Seiji Yano, Roy S. Herbst and Saburo Sone : In vitro and in vivo assays for the proliferative and vascular permeabilization activities of vascular endothelial growth factor(VEGF) and its receptor, Methods in Molecular Medicine, 2003.
Thymidine phosphorylase (TP) is a rate-limiting enzyme in the thymidine catabolic pathway. TP is identical to platelet-derived endothelial cell growth factor and contributes to tumour angiogenesis. TP induces the generation of reactive oxygen species (ROS) and enhances the expression of oxidative stress-responsive genes, such as interleukin (IL)-8. However, the mechanism underlying ROS induction by TP remains unclear. In the present study, we demonstrated that TP promotes NADPH oxidase-derived ROS signalling in cancer cells. NADPH oxidase inhibition using apocynin or small interfering RNAs (siRNAs) abrogated the induction of IL-8 and ROS in TP-expressing cancer cells. Meanwhile, thymidine catabolism induced by TP increased the levels of NADPH and intermediates of the pentose phosphate pathway (PPP). Both siRNA knockdown of glucose 6-phosphate dehydrogenase (G6PD), a rate-limiting enzyme in PPP, and a G6PD inhibitor, dihydroepiandrosterone, reduced TP-induced ROS production. siRNA downregulation of 2-deoxy-D-ribose 5-phosphate (DR5P) aldolase, which is needed for DR5P to enter glycolysis, also suppressed the induction of NADPH and IL-8 in TP-expressing cells. These results suggested that TP-mediated thymidine catabolism increases the intracellular NADPH level via the PPP, which enhances the production of ROS by NADPH oxidase and activates its downstream signalling.
Hisatsugu Goto, Okano Yoshio, Machida Hisanori, Hatakeyama Nobuo, Ogushi Fumitaka, Haku Takashi, Takanori Kanematsu, Tomoyuki Urata, Souji Kakiuchi, Masaki Hanibuchi, Saburo Sone and Yasuhiko Nishioka : Phase II study of tailored S-1 monotherapy with a 1-week interval after a 2-week dosing period in elderly patients with advanced non-small cell lung cancer., Respiratory Investigation, Vol.56, No.1, 80-86, 2018.
(要約)
S-1 is an oral fluoropyrimidine that is active in the treatment of non-small cell lung cancer (NSCLC); however, an optimal treatment schedule and appropriate dose adjustments of S-1 in elderly patients have not yet been established. We conducted a phase II trial to evaluate the efficacy and safety of a 2-week S-1 monotherapy treatment followed by a 1-week interval as a first-line treatment of elderly NSCLC patients, by adjusting the dose based on the individual creatinine clearance (Ccr) and body surface area (BSA). The primary endpoint was the disease control rate. Forty patients were enrolled. The disease control and response rates were 89.5% (95% confidence interval [CI] = 79.8-99.2) and 7.9% (95% CI = 0.0-16.4), respectively. The median progression-free survival and overall survival times were 4.4 months (95% CI = 4.2-8.5) and 17.0 months (95% CI = 11.2-18.7), respectively. Neutropenia, anorexia, hyponatremia, hypokalemia, and pneumonia of grade ≥ 3 occurred in 5.0%, 7.5%, 5.0%, 2.5%, and 2.5% of patients, respectively. Among the patient-reported outcomes, most of the individual factors in the patients' quality of life, including upper intestine-related symptoms improved with the treatment, except for dyspnea, which slightly albeit continuously worsened throughout the study. In elderly patients with previously untreated advanced NSCLC, a 2-week S-1 monotherapy treatment, tailored to both the Ccr and BSA, with a 1-week interval was well tolerated and demonstrated promising efficacy. This study was registered at the University Hospital Medical Information Network (UMIN) Center (ID: UMIN000002035), Japan.
(キーワード)
Administration, Oral / Aged / Aged, 80 and over / Antimetabolites, Antineoplastic / Body Surface Area / Carcinoma, Non-Small-Cell Lung / Creatinine / Drug Administration Schedule / Drug Combinations / Female / Humans / Lung Neoplasms / Male / Metabolic Clearance Rate / Oxonic Acid / Precision Medicine / Survival Rate / Tegafur / Treatment Outcome
Thymidine phosphorylase (TP), a rate-limiting enzyme in thymidine catabolism, plays a pivotal role in tumor progression; however, the mechanisms underlying this role are not fully understood. Here, we found that TP-mediated thymidine catabolism could supply the carbon source in the glycolytic pathway and thus contribute to cell survival under conditions of nutrient deprivation. In TP-expressing cells, thymidine was converted to metabolites, including glucose 6-phosphate, lactate, 5-phospho-α-D-ribose 1-diphosphate, and serine, via the glycolytic pathway both in vitro and in vivo. These thymidine-derived metabolites were required for the survival of cells under low-glucose conditions. Furthermore, activation of thymidine catabolism was observed in human gastric cancer. These findings demonstrate that thymidine can serve as a glycolytic pathway substrate in human cancer cells.
Taisuke Matsuo, Le Tan Dat, Masato Komatsu, Tetsuro Yoshimaru, Kei Daizumoto, Saburo Sone, Yasuhiko Nishioka and Toyomasa Katagiri : Early growth response 4 is involved in cell proliferation of small cell lung cancer through transcriptional activation of its downstream genes., PLoS ONE, Vol.9, No.11, e113606, 2014.
(要約)
Small cell lung cancer (SCLC) is aggressive, with rapid growth and frequent bone metastasis; however, its detailed molecular mechanism remains poorly understood. Here, we report the critical role of early growth factor 4 (EGR4), a DNA-binding, zinc-finger transcription factor, in cell proliferation of SCLC. EGR4 overexpression in HEK293T cells conferred significant upregulation of specific splice variants of the parathyroid hormone-related protein (PTHrP) gene, resulting in enhancement of the secretion of PTHrP protein, a known mediator of osteolytic bone metastasis. More importantly, depletion of EGR4 expression by siRNA significantly suppressed growth of the SCLC cell lines, SBC-5, SBC-3 and NCI-H1048. On the other hand, introduction of EGR4 into NIH3T3 cells significantly enhanced cell growth. We identified four EGR4 target genes, SAMD5, RAB15, SYNPO and DLX5, which were the most significantly downregulated genes upon depletion of EGR4 expression in all of the SCLC cells examined, and demonstrated the direct recruitment of EGR4 to their promoters by ChIP and luciferase reporter analysis. Notably, knockdown of the expression of these genes by siRNA remarkably suppressed the growth of all the SCLC cells. Taken together, our findings suggest that EGR4 likely regulates the bone metastasis and proliferation of SCLC cells via transcriptional regulation of several target genes, and may therefore be a promising target for the development of anticancer drugs for SCLC patients.
Yuko Toyoda, Sho Tabata, Jun Kishi, Takuya Kuramoto, Atsushi Mitsuhashi, Atsuro Saijo, Hiroshi Kawano, Hisatsugu Goto, Yoshinori Aono, Masaki Hanibuchi, Hideaki Horikawa, Toshihiro Nakajima, Tatsuhiko Furukawa, Saburo Sone, Shin-ichi Akiyama and Yasuhiko Nishioka : Thymidine Phosphorylase Regulates the Expression of CXCL10 in Rheumatoid Arthritis Fibroblast-like Synoviocytes, Arthritis & Rheumatology, Vol.66, No.3, 560-568, 2014.
(要約)
Thymidine phosphorylase (TP) in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) is induced by tumor necrosis factor α (TNFα) and other cytokines that have been reported to be major inflammation mediators in RA. We previously demonstrated that TP plays an important role in angiogenesis and tumor growth, invasion, and metastasis. The aim of this study was to investigate whether the role of TP in the pathogenesis of RA is similar to its role in tumors. In FLS obtained from 2 patients with RA, the expression of TP, interferon-γ (IFNγ)-inducible protein 10 (CXCL10), and other cytokines was measured by quantitative real-time polymerase chain reaction, immunoblotting, and enzyme-linked immunosorbent assays. Microarray analysis was performed using FLS transfected with TYMP complementary DNA and treated with a TP inhibitor. The expression of TP in FLS was up-regulated by TNFα, interleukin-1β (IL-1β), IL-17, IFNγ, and lipopolysaccharide. Microarray analysis of FLS overexpressing TP identified CXCL10 as a thymidine phosphorylase-related gene. The expression of CXCL10 was induced by TNFα, and this induction was suppressed by TYMP small interfering RNA and TP inhibitor. Furthermore, the combination of TNFα and IFNγ synergistically augmented the expression of TP and CXCL10. TP-induced CXCL10 expression was suppressed by the antioxidant EUK-8. In the synovial tissue of patients with RA, TP levels were significantly correlated with CXCL10 expression. The combination of TNFα and IFNγ strongly induced the expression of thymidine phosphorylase in RA FLS. The induction of thymidine phosphorylase enhanced the expression of CXCL10, which may contribute to the Th1 phenotype and bone destruction observed in RA.
Katsuhiro Kinoshita, Yoshinori Aono, Momoyo Azuma, Jun Kishi, Akio Takezaki, Masami Kishi, Hideki Makino, Hiroyasu Okazaki, Hisanori Uehara, Keisuke Izumi, Saburo Sone and Yasuhiko Nishioka : Antifibrotic effects of focal adhesion kinase inhibitor in bleomycin-induced pulmonary fibrosis in mice., American Journal of Respiratory Cell and Molecular Biology, Vol.49, No.4, 536-543, 2013.
(要約)
Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase involved in various biological functions, including cell survival, proliferation, migration, and adhesion. FAK is an essential factor for transforming growth factor to induce myofibroblast differentiation. In the present study, we investigated whether the targeted inhibition of FAK by using a specific inhibitor, TAE226, has the potential to regulate pulmonary fibrosis. TAE226 showed inhibitory activity of autophosphorylation of FAK at tyrosine 397 in lung fibroblasts. The addition of TAE226 inhibited the proliferation of lung fibroblasts in response to various growth factors, including platelet-derived growth factor and insulin-like growth factor I, in vitro. TAE226 strongly suppressed the production of type I collagen by lung fibroblasts. Furthermore, treatment of fibroblasts with TAE226 reduced the expression of -smooth muscle actin induced by transforming growth factor , indicating the inhibition of differentiation of fibroblasts to myofibroblasts. Administration of TAE226 ameliorated the pulmonary fibrosis induced by bleomycin in mice even when used late in the treatment. The number of proliferating mesenchymal cells was reduced in the lungs of TAE226-treated mice. These data suggest that FAK signal plays a significant role in the progression of pulmonary fibrosis and that it can become a promising target for therapeutic approaches to pulmonary fibrosis.
Shinji Abe, Yuki Morita, Mika Kato Kaneko, Masaki Hanibuchi, Yuta Tsujimoto, Hisatsugu Goto, Souji Kakiuchi, Yoshinori Aono, Jun Huang, Seidai Sato, Masatoshi Kishuku, Yuki Taniguchi, Mami Azuma, Kazuyoshi Kawazoe, Yoshitaka Sekido, Seiji Yano, Shin-ichi Akiyama, Saburo Sone, Kazuo Minakuchi, Yukinari Kato and Yasuhiko Nishioka : A novel targeting therapy of malignant mesothelioma using anti-podoplanin antibody, The Journal of Immunology, Vol.190, No.12, 6239-6249, 2013.
(要約)
Podoplanin (Aggrus), which is a type I transmembrane sialomucin-like glycoprotein, is highly expressed in malignant pleural mesothelioma (MPM). We previously reported the generation of a rat anti-human podoplanin Ab, NZ-1, which inhibited podoplanin-induced platelet aggregation and hematogenous metastasis. In this study, we examined the antitumor effector functions of NZ-1 and NZ-8, a novel rat-human chimeric Ab generated from NZ-1 including Ab-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity against MPM in vitro and in vivo. Immunostaining with NZ-1 showed the expression of podoplanin in 73% (11 out of 15) of MPM cell lines and 92% (33 out of 36) of malignant mesothelioma tissues. NZ-1 could induce potent ADCC against podoplanin-positive MPM cells mediated by rat NK (CD161a(+)) cells, but not murine splenocytes or human mononuclear cells. Treatment with NZ-1 significantly reduced the growth of s.c. established tumors of MPM cells (ACC-MESO-4 or podoplanin-transfected MSTO-211H) in SCID mice, only when NZ-1 was administered with rat NK cells. In in vivo imaging, NZ-1 efficiently accumulated to xenograft of MPM, and its accumulation continued for 3 wk after systemic administration. Furthermore, NZ-8 preferentially recognized podoplanin expressing in MPM, but not in normal tissues. NZ-8 could induce higher ADCC mediated by human NK cells and complement-dependent cytotoxicity as compared with NZ-1. Treatment with NZ-8 and human NK cells significantly inhibited the growth of MPM cells in vivo. These results strongly suggest that targeting therapy to podoplanin with therapeutic Abs (i.e., NZ-8) derived from NZ-1 might be useful as a novel immunotherapy against MPM.
Atsushi Mitsuhashi, Hisatsugu Goto, Takuya Kuramoto, Sho Tabata, Sawaka Yukishige, Shinji Abe, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, G Julie Ledford, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses lung cancer progression by regulating the polarization of tumor-associated macrophages., The American Journal of Pathology, Vol.182, No.5, 1843-1853, 2013.
(要約)
Surfactant protein A (SP-A) is a large multimeric protein found in the lungs. In addition to its immunoregulatory function in infectious respiratory diseases, SP-A is also used as a marker of lung adenocarcinoma. Despite the finding that SP-A expression levels in cancer cells has a relationship with patient prognosis, the function of SP-A in lung cancer progression is unknown. We investigated the role of SP-A in lung cancer progression by introducing the SP-A gene into human lung adenocarcinoma cell lines. SP-A gene transduction suppressed the progression of tumor in subcutaneous xenograft or lung metastasis mouse models. Immunohistochemical analysis showed that the number of M1 antitumor tumor-associated macrophages (TAMs) was increased and the number of M2 tumor-promoting TAMs was not changed in the tumor tissue produced by SP-A-expressing cells. In addition, natural killer (NK) cells were also increased and activated in the SP-A-expressing tumor. Moreover, SP-A did not inhibit tumor progression in mice depleted of NK cells. Taking into account that SP-A did not directly activate NK cells, these results suggest that SP-A inhibited lung cancer progression by recruiting and activating NK cells via controlling the polarization of TAMs.
Hideki Makino, Yoshinori Aono, Momoyo Azuma, Masami Kishi, Yuki Yokota, Katsuhiro Kinoshita, Akio Takezaki, Jun Kishi, Hiroshi Kawano, Hirohisa Ogawa, Hisanori Uehara, Keisuke Izumi, Saburo Sone and Yasuhiko Nishioka : Antifibrotic effects of CXCR4 antagonist in bleomycin-induced pulmonary fibrosis in mice., The Journal of Medical Investigation : JMI, Vol.60, No.1-2, 127-137, 2013.
(要約)
Circulating fibrocytes had been reported to migrate into the injured lungs, and contribute to fibrogenesis via chemokine-chemokine receptor systems including CXCL12-CXCR4 axis. Here we hypothesized that blockade of CXCR4 might inhibit the migration of fibrocytes to the injured lungs and the subsequent pulmonary fibrosis. To explore the antifibrotic effects of blockade of CXCR4, we used a specific antagonist for CXCR4, AMD3100, in bleomycin-induced pulmonary fibrosis model in mice. Administration of AMD3100 significantly improved the loss of body weight of mice treated with bleomycin, and inhibited the fibrotic lesion in subpleural areas of the lungs. The quantitative analysis demonstrated that treatment with AMD3100 reduced the collagen content and fibrotic score (Aschcroft score) in the lungs. Although AMD3100 did not affect cell classification in bronchoalveolar lavage fluid on day 7, the percentage of lymphocytes was reduced by AMD3100 on day 14. AMD3100 directly inhibited the migration of human fibrocytes in response to CXCL12 in vitro, and reduced the trafficking of fibrocytes into the lungs treated with bleocmycin in vivo. These results suggest that the blockade of CXCR4 might be useful strategy for therapy of patients with pulmonary fibrosis via inhibiting the migration of circulating fibrocytes.
T Yamada, S Takeuchi, N Fujita, A Nakamura, W Wang, Q Li, M Oda, T Mitsudomi, Y Yatabe, Y Sekido, J Yoshida, M Higashiyama, M Noguchi, H Uehara, Yasuhiko Nishioka, Saburo Sone and Seiji Yano : Akt kinase-interacting protein1, a novel therapeutic target for lung cancer with EGFR-activating and gatekeeper mutations, Oncogene, Vol.32, No.37, 4427-4435, 2012.
(要約)
Despite initial dramatic response, epidermal growth factor receptor (EGFR) mutant lung cancer patients always acquire resistance to EGFR-tyrosine kinase inhibitors (TKIs). Gatekeeper T790M mutation in EGFR is the most prevalent genetic alteration underlying acquired resistance to EGFR-TKI, and EGFR mutant lung cancer cells are reported to be addictive to EGFR/Akt signaling even after acquired T790M mutation. Here, we focused on Akt kinase-interacting protein1 (Aki1), a scaffold protein of PI3K (phosphoinositide 3-kinase)/PDK1 (3-phosphoinositide-dependent protein kinase)/Akt that determines receptor signal selectivity for non-mutated EGFR, and assessed its role in EGFR mutant lung cancer with or without gatekeeper T790M mutation. Cell line-based assays showed that Aki1 constitutively associates with mutant EGFR in lung cancer cells with (H1975) or without (PC-9 and HCC827) T790M gatekeeper mutation. Silencing of Aki1 induced apoptosis of EGFR mutant lung cancer cells. Treatment with Aki1 siRNA dramatically inhibited growth of H1975 cells in a xenograft model. Moreover, silencing of Aki1 further potentiated growth inhibitory effect of new generation EGFR-TKIs against H1975 cells in vitro. Aki1 was frequently expressed in tumor cells of EGFR mutant lung cancer patients (53/56 cases), including those with acquired resistance to EGFR-TKI treatment (7/7 cases). Our data suggest that Aki1 may be a critical mediator of survival signaling from mutant EGFR to Akt, and may therefore be an ideal target for EGFR mutant lung cancer patients, especially those with acquired EGFR-TKI resistance due to EGFR T790M gatekeeper mutation.
Seidai Satou, Masaki Hanibuchi, Takuya Kuramoto, Nodoka Yamamori, Hisatsugu Goto, Hirohisa Ogawa, Atsushi Mitsuhashi, The Trung Van, Souji Kakiuchi, Shin-ichi Akiyama, Yasuhiko Nishioka and Saburo Sone : Macrophage stimulating protein promotes liver metastases of small cell lung cancer cells by affecting the organ microenvironment., Clinical & Experimental Metastasis, Vol.30, No.3, 333-344, 2012.
(要約)
The organ microenvironment significantly affects the processes of cancer metastasis. Elucidating the molecular mechanisms of interaction between tumor cells and the organ microenvironment is crucial for the development of effective therapeutic strategies to eradicate cancer metastases. Macrophage stimulating protein (MSP), an activator of macrophages, regulates a pleiotropic array of effects, including proliferation, cellular motility, invasiveness, angiogenesis, and resistance to anoikis. However, the role of MSP in cancer metastasis is still largely unknown. In this study, the action of MSP on the production of metastases was determined in a multiple-organ metastasis model. The murine MSP gene was transfected into two human SCLC cell lines, SBC-5 and H1048, to establish transfectants secreting biologically active MSP. MSP gene transduction did not affect cell proliferation and motility in vitro. Intravenously inoculated MSP transfectants produced significantly larger numbers of liver metastases than parental cells or vector control clones, while there were no significant differences in bone or lung metastases among them. Immunohistochemical analyses of liver metastases revealed that tumor-associated microvessel density and tumor-infiltrating macrophages were significantly increased in lesions produced by MSP transfectants. MSP could stimulate the migration of murine macrophages and endothelial cells in vitro. Consequently, MSP may be one of the major determinants that affects the properties of tumor stroma and that produces a permissive microenvironment to promote cancer metastasis.
Takuya Kuramoto, Hisatsugu Goto, Atsushi Mitsuhashi, Sho Tabata, Hirohisa Ogawa, Hisanori Uehara, Atsuro Saijo, Souji Kakiuchi, Yoichi Maekawa, Koji Yasutomo, Masaki Hanibuchi, Shin-ichi Akiyama, Saburo Sone and Yasuhiko Nishioka : Dll4-Fc, an inhibitor of Dll4-Notch signaling, suppresses liver metastasis of small cell lung cancer cells through down-regulation of NF-kappa-B activity, Molecular Cancer Therapeutics, Vol.11, No.12, 2578-2587, 2012.
(要約)
Notch signaling regulates cell-fate decisions during development and postnatal life. Little is known, however, about the role of Delta-like-4 (Dll4)-Notch signaling between cancer cells, or how this signaling affects cancer metastasis. We, therefore, assessed the role of Dll4-Notch signaling in cancer metastasis. We generated a soluble Dll4 fused to the IgG1 constant region (Dll4-Fc) that acts as a blocker of Dll4-Notch signaling and introduced it into human small cell lung cancer (SCLC) cell lines expressing either high levels (SBC-3 and H1048) or low levels (SBC-5) of Dll4. The effects of Dll4-Fc on metastasis of SCLC were evaluated using a mouse model. Although Dll4-Fc had no effect on the liver metastasis of SBC-5, the number of liver metastasis inoculated with SBC-3 and H1048 cells expressing Dll4-Fc was significantly lower than that injected with control cells. To study the molecular mechanisms of the effects of Dll4-Fc on liver metastasis, a PCR array analysis was conducted. Because the expression of NF-κB target genes was affected by Dll4-Fc, we conducted an electrophoretic mobility shift assay and observed that NF-κB activities, both with and without stimulation by TNF-α, were downregulated in Dll4-Fc-overexpressing SBC-3 and H1048 cells compared with control cells. Moreover, Dll4-Fc attenuates, at least in part, the classical and alternative NF-κB activation pathway by reducing Notch1 signaling. These results suggest that Dll4-Notch signaling in cancer cells plays a critical role in liver metastasis of SCLC by regulating NF-κB signaling.
Trung The Van, Masaki Hanibuchi, Hisatsugu Goto, Takuya Kuramoto, Sawaka Yukishige, Souji Kakiuchi, Seidai Sato, Satoshi Sakaguchi, Le Tan Dat, Yasuhiko Nishioka, Shin-ichi Akiyama and Saburo Sone : SU6668, a multiple tyrosine kinase inhibitor, inhibits the progression of human malignant pleural mesothelioma in an orthotopic model, Respirology, Vol.17, No.6, 984-990, 2012.
(要約)
Malignant pleural mesothelioma (MPM) is an aggressive neoplasm of the mesothelium with high chemotherapeutic resistance. In this study, the preclinical therapeutic activity of the multiple tyrosine kinase inhibitor, SU6668, against MPM was examined. Two human MPM cell lines with different pro-angiogenic cytokine expression, Y-MESO-14 cells that express high levels of vascular endothelial growth factor (VEGF) and MSTO-211H cells that express high levels of basic fibroblast growth factor (bFGF), were orthotopically inoculated into the thoracic cavities of mice with severe combined immunodeficiency. The mice with MPM were treated or not treated with SU6668 (200 mg/kg/day). SU6668 abrogated the proliferation of endothelial cells stimulated by VEGF or bFGF, but did not directly affect the growth of human MPM cells in vitro. In this orthotopic implantation model, treatment with SU6668 effectively reduced tumour weight and pleural effusion volumes, in association with inhibition of the growth of tumour vasculature. More importantly, treatment with SU6668 significantly prolonged survival time in mice with MPM. These findings suggest that SU6668 has a promising therapeutic effect on the progression of MPM in vivo through its anti-angiogenic effects.
Shinji Takeuchi, Wei Wang, Qi Li, Tadaaki Yamada, Kenji Kita, S Ivan Donev, Takahiro Nakamura, Kunio Matsumoto, Eiji Shimizu, Yasuhiko Nishioka, Saburo Sone, Takayuki Nakagawa, Toshimitsu Uenaka and Seiji Yano : Dual inhibition of Met kinase and angiogenesis to overcome HGF-induced EGFR-TKI resistance in EGFR mutant lung cancer., The American Journal of Pathology, Vol.181, No.3, 1034-1043, 2012.
(要約)
Acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) is a serious problem in the management of EGFR mutant lung cancer. We recently reported that hepatocyte growth factor (HGF) induces resistance to EGFR-TKIs by activating the Met/PI3K pathway. HGF is also known to induce angiogenesis in cooperation with vascular endothelial growth factor (VEGF), which is an important therapeutic target in lung cancer. Therefore, we hypothesized that dual inhibition of HGF and VEGF may be therapeutically useful for controlling HGF-induced EGFR-TKI-resistant lung cancer. We found that a dual Met/VEGF receptor 2 kinase inhibitor, E7050, circumvented HGF-induced EGFR-TKI resistance in EGFR mutant lung cancer cell lines by inhibiting the Met/Gab1/PI3K/Akt pathway in vitro. HGF stimulated VEGF production by activation of the Met/Gab1 signaling pathway in EGFR mutant lung cancer cell lines, and E7050 showed an inhibitory effect. In a xenograft model, tumors produced by HGF-transfected Ma-1 (Ma-1/HGF) cells were more angiogenic than vector control tumors and showed resistance to gefitinib. E7050 alone inhibited angiogenesis and retarded growth of Ma-1/HGF tumors. E7050 combined with gefitinib induced marked regression of tumor growth. Moreover, dual inhibition of HGF and VEGF by neutralizing antibodies combined with gefitinib also markedly regressed tumor growth. These results indicate the therapeutic rationale of dual targeting of HGF-Met and VEGF-VEGF receptor 2 for overcoming HGF-induced EGFR-TKI resistance in EGFR mutant lung cancer.
Sho Tabata, Ryuji Ikeda, Masatatu Yamamoto, Tatsuhiko Furukawa, Takuya Kuramoto, Yasuo Takeda, Katsushi Yamada, Misako Haraguchi, Yasuhiko Nishioka, Saburo Sone and Shin-ichi Akiyama : Thymidine phosphorylase enhances reactive oxygen species generation and interleukin-8 expression in human cancer cells., Oncology Reports, Vol.28, No.3, 895-902, 2012.
(要約)
Thymidine phosphorylase (TP) is an angiogenic factor that plays a pivotal role in tumor angiogenesis. Various kinds of solid tumors express TP and high TP activity is correlated with microvessel density. We have previously reported that TP enhances interleukin-8 (IL-8) expression in KB human epidermoid carcinoma cells. In this study, TP was shown to be involved in enhanced expression of IL-8 in EJ human bladder cancer cells and Yumoto human cervical cancer cells as well as KB human epidermoid carcinoma cells. The enzymatic activity of TP was required for the enhanced expression of IL-8. A degradation product of thymidine was implicated in the enhanced expression of IL-8. TP augmented reactive oxygen species (ROS) generation in KB and Yumoto cells, and the enzymatic activity of TP was again required for the generation of ROS. An antioxidant, N-acetylcysteine (NAC), attenuated the generation of ROS and IL-8 mRNA expression in KB and Yumoto cells, and H2O2 increased IL-8 mRNA expression in Yumoto cells, suggesting that ROS generated by TP caused the increased expression of IL-8 mRNA. Since TP also reduced cellular glutathione levels and transcription of γ-GCS in KB cells, the TP-induced augmentation of ROS may be partially attributed to the decreased glutathione. Our findings suggest that thymidine-derived sugars enhanced ROS generation and consequently increased IL-8 expression.
Adel Gomaa Mohammed Gabr, Hisatsugu Goto, Masaki Hanibuchi, Hirohisa Ogawa, Takuya Kuramoto, Minako Suzuki, Atsuro Saijo, Souji Kakiuchi, Van The Trung, Satoshi Sakaguchi, Yoichiro Moriya, Saburo Sone and Yasuhiko Nishioka : Erlotinib prevents experimental metastases of human small cell lung cancer cells with no epidermal growth factor receptor expression, Clinical & Experimental Metastasis, Vol.29, No.3, 207-216, 2012.
(要約)
Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor (EGFR) gene. On the other hand, some lung cancer patients with wild type EGFR also respond to EGFR-TKIs, suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells. However, the effect of EGFR-TKIs on host microenvironments is largely unknown. A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells. This model was used to investigate the therapeutic efficacy of erlotinib, an EGFR-TKI, on multiple organ metastases induced by human small cell lung cancer cells (SBC-5 cells) that did not express EGFR. Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro, it significantly suppressed bone and lung metastases in vivo, but not liver metastases. An immunohistochemical analysis revealed that, erlotinib significantly suppressed the number of osteoclasts in bone metastases, whereas no difference was seen in microvessel density. Moreover, erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line (MC3T3-E1 cells). These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells.
Yoshinori Aono, Julie G. Ledford, Sambuddho Mukherjee, Hirohisa Ogawa, Yasuhiko Nishioka, Saburo Sone, Michael F. Beers, Paul W. Noble and Jo Rae Wright : Surfactant protein-D regulates effector cell function and fibrotic lung remodeling in response to bleomycin injury., American Journal of Respiratory and Critical Care Medicine, Vol.185, No.5, 525-536, 2012.
(要約)
Surfactant protein (SP)-D and SP-A have been implicated in immunomodulation in the lung. It has been reported that patients with idiopathic pulmonary fibrosis (IPF) often have elevated serum levels of SP-A and SP-D, although their role in the disease is not known. The goal of this study was to test the hypothesis that SP-D plays an important role in lung fibrosis using a mouse model of fibrosis induced by bleomycin (BLM). Triple transgenic inducible SP-D mice (iSP-D mice), in which rat SP-D is expressed in response to doxycycline (Dox) treatment, were administered BLM (100 U/kg) or saline subcutaneously using miniosmotic pumps. BLM-treated iSP-D mice off Dox (SP-D off) had increased lung fibrosis compared with mice on Dox (SP-D on). SP-D deficiency also increased macrophage-dominant cell infiltration and the expression of profibrotic cytokines (transforming growth factor [TGF]-1, platelet-derived growth factor-AA). Alveolar macrophages isolated from BLM-treated iSP-D mice off Dox (SP-D off) secreted more TGF-1. Fibrocytes, which are bone marrow-derived mesenchymal progenitor cells, were increased to a greater extent in the lungs of the BLM-treated iSP-D mice off Dox (SP-D off). Fibrocytes isolated from BLM-treated iSP-D mice off Dox (SP-D off) expressed more of the profibrotic cytokine TGF-1 and more CXCR4, a chemokine receptor that is important in fibrocyte migration into the lungs. Exogenous SP-D administered intratracheally attenuated BLM-induced lung fibrosis in SP-D(-/-) mice. These data suggest that alveolar SP-D regulates numbers of macrophages and fibrocytes in the lungs, profibrotic cytokine expression, and fibrotic lung remodeling in response to BLM injury.
Hirohisa Ogawa, Masahiko Azuma, Hisanori Uehara, Tetsuyuki Takahashi, Yasuhiko Nishioka, Saburo Sone and Keisuke Izumi : Nerve growth factor derived from bronchial epithelium afer chronic mite antigen exposure contributes to airway hyperresponsiveness by inducing hyperinnervaiton, and is inhibited by in vivo siRNA., Clinical and Experimental Allergy, Vol.42, No.3, 460-470, 2012.
(要約)
Bronchial asthma is a chronic allergic airway inflammatory disease. Neurotrophins, including nerve growth factor (NGF), play an important role in the pathogenesis of asthma. However, the effects of NGF derived from epithelium on airway hyperresponsiveness (AHR) after antigen sensitization/exposure remain uncertain. In this study, we examined the role of NGF on AHR after chronic antigen exposure and the effect of inhibiting NGF by in vivo siRNA on AHR exacerbation. We generated chronic mouse models of bronchial asthma using house-dust mite antigen (Dermatophagoides pteronyssinus; Dp). NGF concentrations in bronchoalveolar lavage fluid (BALF), lung histopathology, hyperresponsiveness, and related neuronal peptides and cytokines in supernatants of lung homogenates were determined. NGF in BALF was increased in a dose- and time-dependent manner, and was expressed primarily in bronchial epithelium. Nerve fibres and substance P-positive fibres were detected in subepithelium of Dp-sensitized and challenged mice over 4 weeks of mite antigen exposure. AHR was positively correlated with NGF concentration and nerve fibre innervation. AHR, modulation of innervation, and increased substance P were inhibited by in vivo administration of siRNA that targeted NGF, although the inhibition of NGF did not affect allergic inflammation and subepithelial fibrosis. These findings suggest that NGF derived from bronchial and alveolar epithelium plays an important role in AHR after chronic exposure to mite antigen. NGF inhibition could potentially manage bronchial asthma, including AHR.
Wei Wang, Qi Li, Shinji Takeuchi, Tadaaki Yamada, Hitomi Koizumi, Takahiro Nakamura, Kunio Matsumoto, Naofumi Mukaida, Yasuhiko Nishioka, Saburo Sone, Takayuki Nakagawa, Toshimitsu Uenaka and Seiji Yano : Met kinase inhibitor E7050 reverses three different mechanisms of hepatocyte growth factor-induced tyrosine kinase inhibitor resistance in EGFR mutant lung cancer., Clinical Cancer Research, Vol.18, No.6, 1663-1671, 2012.
(要約)
Hepatocyte growth factor (HGF) induces resistance to reversible and irreversible epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKI) in EGFR mutant lung cancer cells by activating Met and the downstream phosphoinositide 3-kinase (PI3K)/Akt pathway. Moreover, continuous exposure to HGF accelerates the emergence of EGFR-TKI-resistant clones. We assayed whether a new Met kinase inhibitor, E7050, which is currently being evaluated in clinical trials, could overcome these three mechanisms of resistance to EGFR-TKIs. The effects of E7050 on HGF-induced resistance to reversible (gefitinib), irreversible (BIBW2992), and mutant-selective (WZ4002) EGFR-TKIs were determined using the EGFR mutant human lung cancer cell lines PC-9 and HCC827 with an exon 19 deletion and H1975 with an T790M secondary mutation. PC-9 cells were mixed with HGF-producing fibroblasts, MRC-5 cells, and subcutaneously inoculated into severe combined immunodeficient mice, and the therapeutic effects of E7050 plus gefitinib were assayed. E7050 circumvented resistance to all of the reversible, irreversible, and mutant-selective EGFR-TKIs induced by exogenous and/or endogenous HGF in EGFR mutant lung cancer cell lines, by blocking the Met/Gab1/PI3K/Akt pathway in vitro. E7050 also prevented the emergence of gefitinib-resistant HCC827 cells induced by continuous exposure to HGF. In the in vivo model, E7050 plus gefitinib resulted in marked regression of tumor growth associated with inhibition of Akt phosphorylation in cancer cells. A new Met kinase inhibitor, E7050, reverses the three HGF-induced mechanisms of gefitinib resistance, suggesting that E7050 may overcome HGF-induced resistance to gefitinib and next-generation EGFR-TKIs.
Keiko Miyake, Kenji Tani, Souji Kakiuchi, Chiyuki Suzuka, Yuko Toyoda, Jun Kishi, Toshifumi Tezuka, Shino Yuasa, Masaki Hanibuchi, Yoshinori Aono, Yasuhiko Nishioka and Saburo Sone : Epidermal growth factor receptor-tyrosine kinase inhibitor (gefitinib) augments pneumonitis, but attenuates lung fibrosis in response to radiation injury in rats, The Journal of Medical Investigation : JMI, Vol.59, No.1-2, 174-185, 2012.
(要約)
Gefitinib, an inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase, has been reported to be associated with interstitial lung disorders, and their high incidence and mortality have become a matter of great concern, especially in Japan. In this study, we investigated the effect of gefitinib on different phases of radiation-induced lung disorders in an experimental model. The thoraxes of Wistar rats were irradiated on day 1 with a single X-ray dose of 20 Gy, and gefitinib (50 mg/kg/day) was orally administered from day 1 to 14. The rat lungs were harvested on days 15 and 57 and the bronchoalveolar lavage (BAL) was performed. Gefitinib treatment increased the infiltration of inflammatory cells, which produced more pro-inflammatory cytokines (IL-6, IL-1), in the lungs of the irradiated rats on days 15 and 57, while gefitinib treatment reduced collagen content of the lungs in irradiated rats and decreased proliferation and EGFR expression in the lung fibroblasts from irradiated rats on day 57. In irradiated rats, gefitinib treatment augmented lung inflammation, including inflammatory cell infiltration and pro-inflammatory cytokine expression, while gefitinib treatment attenuated fibrotic lung remodeling due to the inhibition of lung fibroblast proliferation.
Le T Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Souji Kakiuchi, Hisatsugu Goto, Masaki Hanibuchi, Takuya Kuramoto, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Identification of genes potentially involved in bone metastasis by genome-wide gene expression profile analysis of non-small cell lung cancer in mice, International Journal of Oncology, Vol.40, No.5, 1455-1469, 2012.
(要約)
Lung cancer is commonly associated with multi-organ metastasis, and the bone is a frequent metastatic site for lung cancer. However, the molecular mechanism of organ-specific metastasis remains poorly understood. To elucidate this issue, we analyzed in this study genome-wide gene expression profiles of 15 metastatic lesions from three organs (bone, lung and liver) in a mouse model with multi-organ metastasis properties of human non-small cell lung cancer cells (ACC-LC319/bone2), using a combination of laser-microbeam microdissection and DNA microarrays. We identified 299 genes that could potentially be involved in the organ-selective nature of lung cancer metastasis. Among them, 77 were bone-specifically expressed elements, including genes involved in cell adhesion, cytoskeleton/cell motility, extracellular matrix remodeling and cell-cell signaling as well as genes already known to be involved in the bone metastasis of breast cancers. Quantitative RT-PCR confirmed the specific upregulation of eight genes in bone metastasis tumors, suggesting that these genes may be involved in bone metastasis. Our findings should be helpful for a better understanding of the molecular aspects of the metastatic process in different organs, and could lead to molecular target-based anticancer drugs and prevention of metastasis, especially bone metastasis.
Tadaaki Yamada, Hideaki Bando, Shinji Takeuchi, Kenji Kita, Qi Li, Wei Wang, Shiro Akinaga, Yasuhiko Nishioka, Saburo Sone and Seiji Yano : Genetically engineered humanized anti-ganglioside GM2 antibody against multiple organ metastasis produced by GM2-expressing small-cell lung cancer cells., Cancer Science, Vol.102, No.12, 2157-2163, 2011.
(要約)
Small-cell lung cancer (SCLC) grows rapidly and metastasizes to multiple organs. We examined the antimetastatic effects of the humanized anti-ganglioside GM2 (GM2) antibodies, BIW-8962 and KM8927, compared with the chimeric antibody KM966, in a SCID mouse model of multiple organ metastases induced by GM2-expressing SCLC cells. BIW-8962 and KM8927 induced higher antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity than KM966 against the GM2-expressing SCLC cell line SBC-3 in vitro. These humanized antibodies inhibited the production of multiple organ metastases, increased the number of apoptotic cells, and prolonged the survival of the SCID mice. Histological analyses using clinical specimens showed that SCLC cells expressed GM2. These findings suggest that humanized anti-GM2 antibodies could be therapeutically useful for controlling multiple organ metastases of GM2-expressing SCLC.
Qi Li, Wei Wang, Tadaaki Yamada, Kunio Matsumoto, Katsuya Sakai, Yoshimi Bando, Hisanori Uehara, Yasuhiko Nishioka, Saburo Sone, Shotaro Iwakiri, Kazumi Itoi, Teruhiro Utsugi, Kazuo Yasumoto and Seiji Yano : Pleural mesothelioma instigates tumor-associated fibroblasts to promote progression via a malignant cytokine network., The American Journal of Pathology, Vol.179, No.3, 1483-1493, 2011.
(要約)
The tumor microenvironment is crucial to the progression of various malignancies. Malignant pleural mesothelioma (MPM), which originates from the pleura, grows aggressively in the thoracic cavity. Here we describe an orthotopic implantation SCID mouse model of MPM and demonstrate that α-SMA-positive fibroblast-like cells accumulate in the tumors produced by the human MPM cell lines MSTO-211H and Y-Meso-14. We assessed the interaction between MPM cells and their microenvironments, focusing on tumor-associated fibroblasts. MSTO-211H and Y-Meso-14 cells produced fibroblast growth factor-2 (FGF-2) and/or platelet-derived growth factor-AA (PDGF-AA); they also enhanced growth, migration, and production of hepatocyte growth factor (HGF) by human lung fibroblast MRC-5 cells. MRC-5 cells stimulated HGF-mediated growth and migration of MSTO-211H and Y-Meso-14 cells in an in vitro coculture system. In the orthotopic model, tumor formation by MSTO-211H and Y-Meso-14 cells was significantly inhibited by TSU-68, an inhibitor of FGF, VEGF, and PDGF receptors; imatinib, an inhibitor of PDGF receptors; and NK4, an antagonist of HGF. Histological analyses of clinical specimens from 51 MPM patients revealed considerable tumor-associated fibroblasts infiltration and expression of HGF, together with FGF-2 or PDGF-AA, in tumors. These findings indicate that MPM instigates tumor-associated fibroblasts, promoting tumor progression via a malignant cytokine network. Regulation of this cytokine network may be therapeutically useful for controlling MPM.
Trung The Van, Masaki Hanibuchi, Souji Kakiuchi, Seidai Sato, Takuya Kuramoto, Hisatsugu Goto, Atsushi Mitsuhashi, Yasuhiko Nishioka, Shin-ichi Akiyama and Saburo Sone : The therapeutic efficacy of S-1 against orthotopically implanted human pleural mesothelioma cells in severe combined immunodeficient mice., Cancer Chemotherapy and Pharmacology, Vol.68, No.2, 497-504, 2011.
(要約)
Malignant pleural mesothelioma (MPM) is a highly lethal neoplasm. S-1 has been developed as a novel oral antineoplastic agent based on the modulation of 5-fluorouracil (5-FU) bioactivity. This study was conducted to investigate the preclinical therapeutic effect of S-1 on MPM. We used three human MPM cell lines, Y-MESO-14, NCI-H290 and MSTO-211H. In vitro proliferation of human MPM cells was determined by MTT assay. Human MPM cells were orthotopically implanted into thoracic cavity of SCID mice. Tumor-bearing mice were treated with S-1 or vehicle. The combination of 5-FU and 5-chloro-2,4-dihydroxypyridine (CDHP) was more effective than 5-FU alone in inhibiting MPM cell proliferation in vitro. This combination was most effective in Y-MESO-14 cells, which co-expressed high protein level of dihydropyrimidine dehydrogenase (DPD) and thymidine phosphorylase (TP). In vivo data showed that treatment with S-1 significantly reduced thoracic tumors and pleural effusion produced by Y-MESO-14 cells. Moreover, treatment with S-1 prolonged the survival of Y-MESO-14 cell-bearing SCID mice. We demonstrated that S-1 was effective for inhibiting the proliferation of MPM cells, particularly with both DPD and TP expressions, suggesting that S-1 might be therapeutically effective for control of MPM.
Rentsenkhand Batmunkh, Yasuhiko Nishioka, Yoshinori Aono, Momoyo Azuma, Katsuhiro Kinoshita, Jun Kishi, Hideki Makino, Masami Kishi, Akio Takezaki and Saburo Sone : CCN6 as a profibrotic mediator that stimulates the proliferation of lung fibroblasts via the integrin β1/focal adhesion kinase pathway., The Journal of Medical Investigation : JMI, Vol.58, No.3-4, 188-196, 2011.
(要約)
Idiopathic pulmonary fibrosis is a progressive and lethal disease of the lung that is characterized by the proliferation of fibroblasts and increased deposition of the extracellular matrix. The CCN6/WISP-3 is a member of the CCN family of matricellular proteins, which consists of six members that are involved in many vital biological functions. However, the regulation of lung fibroblasts mediated by CCN6 protein has not been fully elucidated. Here, we demonstrated that CCN6 induced the proliferation of lung fibroblasts by binding to integrin β1, leading to the phosphorylation of FAK(Y397). Furthermore, CCN6 showed a weak, but significant, ability to stimulate the expression of fibronectin. CCN6 was highly expressed in the lung tissues of mice treated with bleomycin. Our results suggest that CCN6 plays a role in the fibrogenesis of the lungs mainly by stimulating the growth of lung fibroblasts and is a potential target for the treatment of pulmonary fibrosis.
Hisatsugu Goto, Masaki Hanibuchi, Satoshi Sakaguchi, Takanori Kanematsu, Souji Kakiuchi, Hideki Tomimoto, Masahiko Azuma, Toshifumi Tezuka, Hiroya Tada, Yukiyo Miki, Toshimi Nakamura, Saburo Sone and Yasuhiko Nishioka : Investigation of the outpatient chemotherapy for lung cancer patients in Tokushima University Hospital., The Journal of Medical Investigation : JMI, Vol.58, No.3-4, 219-226, 2011.
(要約)
Platinum-doublet regimens and docetaxel as first- and second-line chemotherapy, respectively, are shown to prolong the survival of lung cancer patients in various randomized phase III studies. However, the evidence for the efficacy of chemotherapy for lung cancer in the clinical practice is still insufficient. In the present study, we investigated the effectiveness and safety of outpatient chemotherapy for lung cancer in the clinical practice. Ninety-four lung cancer cases were retrospectively analyzed. Among these cases, 67 (71.3%) were non-small cell lung cancer (NSCLC) and 27 (28.7%) were small cell lung cancer (SCLC). The response rates in SCLC and NSCLC patients were 55.6% (15/27) and 16.9% (11/65), respectively. Objective tumor response rates for the patients were found to decrease substantially with each line of treatment as described previously. All adverse events were well tolerated and no treatment-related death was observed. Median time to treatment failures (TTFs) of first-line treatment were 10.1 months and 4.8 months in SCLC and NSCLC, respectively. These findings indicate that even in the setting of clinical practice, the efficacy and safety of chemotherapy is strictly insured by the appropriate therapeutic management.
(キーワード)
Adult / Aged / Aged, 80 and over / Antineoplastic Agents / Carcinoma, Non-Small-Cell Lung / Carcinoma, Small Cell / Female / Hospitals, University / Humans / Lung Neoplasms / Male / Middle Aged / Outpatients / Retrospective Studies
Ivan S. Donev, Wei Wang, Tadaaki Yamada, Qi Li, Shinji Takeuchi, Kunio Matsumoto, Takao Yamori, Yasuhiko Nishioka, Saburo Sone and Seiji Yano : Transient PI3K inhibition induces apoptosis and overcomes HGF-mediated resistance to EGFR-TKIs in EGFR mutant lung cancer., Clinical Cancer Research, Vol.17, No.8, 2260-2269, 2011.
(要約)
Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI), such as gefitinib and erlotinib, show favorable response to EGFR mutant lung cancer. However, the responders acquire resistance almost without exception. We recently reported that hepatocyte growth factor (HGF) induces EGFR-TKI resistance by activating MET that restores downstream mitogen activated protein kinase (MAPK)/extracellular signal regulated kinase (ERK)1/2 and phosphoinositide 3-kinase (PI3K)/Akt signaling. The purpose of this study was to determine whether inhibition of PI3K, a downstream molecule of both EGFR and MET, could overcome HGF-mediated EGFR-TKI resistance in EGFR mutant lung cancer cells PC-9 and HCC827. We explored therapeutic effect of a class I PI3K inhibitor PI-103 on HGF-induced EGFR-TKI resistance in vitro and in vivo. Unlike gefitinib or erlotinib, continuous exposure with PI-103 inhibited proliferation of PC-9 and HCC827 cells, even in the presence of HGF. On the other hand, in gefitinib-resistant xenograft model by using PC-9 cells mixed with HGF high producing fibroblasts, PI-103 monotherapy did not inhibit tumor growth. However, PI-103 combined with gefitinib successfully regressed gefitinib-resistant tumor. In vitro experiments by considering short half-life of PI-103 reveal that transient exposure of PI-103 combined with gefitinib caused sustained inhibition of Akt phosphorylation, but not ERK1/2 phosphorylation, resulting in induction of tumor cell apoptosis even in the presence of HGF. These results indicate that transient blockade of PI3K/Akt pathway by PI-103 and gefitinib could overcome HGF-mediated resistance to EGFR-TKIs by inducing apoptosis in EGFR mutant lung cancer.
Hideki Tomimoto, Masaki Hanibuchi, Fumitaka Ogushi, Yoshio Okano, Tsutomu Shinohara, Hiroyuki Doi, Akiyoshi Yamamoto, Eiji Takeuchi, Akihiko Yamamoto, Masahiko Azuma, Hiroya Tada, Takanori Kanematsu, Souji Kakiuchi, Hisatsugu Goto, Seiji Yano, Yasuhiko Nishioka and Saburo Sone : A multi-institutional phase II study of combination chemotherapy with S-1 plus cisplatin in patients with advanced non-small cell lung cancer., Oncology Letters, Vol.2, No.3, 465-470, 2011.
(要約)
S-1 is an oral anticancer fluoropyrimidine agent designed to elevate anticancer activity with a decrease in gastrointestinal toxicity. We conducted a phase II study to evaluate the efficacy and safety of combination chemotherapy with S-1 plus cisplatin in patients with advanced non-small cell lung cancer (NSCLC). Chemotherapy-naïve patients were treated with S-1 administered orally at 40 mg/m(2) twice a day for 21 consecutive days, and cisplatin (60 mg/m(2)) infused intravenously on day 8, repeated every 5 weeks. Of the 44 patients enrolled in the study, 40 were assessable for efficacy and safety. The median number of cycles administered was 3 (range 1-9 cycles). Among the 40 assessable patients, 7 partial responses were observed, with an overall response rate (RR) of 17.5% [95% confidence interval (CI), 5.2-29.8]. Patients with squamous cell carcinoma showed a significantly higher RR (55.5%) than those with adenocarcinoma (9.1%) or other types of NSCLC (0%). The median progression-free survival was 4.3 months (95% CI, 3.4-4.9), the median survival time was 17.9 months (95% CI, 15.0-20.8), and the 1- and 2-year survival rates were 63.3 and 27.3%, respectively. Major grade 3-4 hematologic toxicities were leukocytopenia (7.5%), neutropenia (5.0%), anemia (15.0%) and thrombocytopenia (2.5%). No grade 4 non-hematologic toxicity or treatment-related death occurred. These results suggest that combination chemotherapy with S-1 plus cisplatin is a promising therapeutic candidate for patients with advanced NSCLC, particularly squamous cell carcinoma.
The effects of the soy isoflavones, genistein, daidzein and equol, on experimental colitis were examined. Equol severely perpetrated dextran sulfate sodium (DSS)-induced colitis as evaluated by the weight loss. Production of the anti-inflammatory cytokine, IL-10, from T cells was decreased in the equol-treated mice. The results show that the soy isoflavone, equol, played an important role in the inflammatory response in the gastrointestinal tract.
BACKGROUND: It is known that endogenously synthesized protoporphyrin IX (PpIX) following the administration of 5-aminolevulinic acid (5-ALA) is an effective photosensitizer for photodynamic diagnosis (PDD). We tested in vivo and in vitro susceptibility of human lung cancer and mesothelioma cells to photodynamic diagnosis (PDD) using 5-aminolevulinic acid (5-ALA) as a photosensitizer. METHODS: Human lung cancer cell lines A549, Ma44-3, FT821 and human mesothelioma cell lines MSTO-211H, NCI-H290, Y-MESO-14 were incubated with 0.03% 5-ALA for 4h. After incubation, protoporphyrin IX (PpIX) fluorescence was detected using a fluorescence microscope. Pleural carcinosis was induced in severe combined immunodeficiency disease mice using the previous cell lines to test the efficacy of PDD in vivo. The mice were sacrificed 4h after oral administration of 400mg/kg of 5-ALA. We counted the visible tumors under white light then fluorescence light. RESULTS: In vitro, clear red fluorescence was observed in all cell lines. The mean fluorescence intensity was stronger in A549 and FT821 cells than Ma44-3 cells (165.59±26.49, 157.62±18.93 vs. 104.01±17.58). Also, MSTO-211H and NCI-H290 cells had stronger fluorescence intensity than Y-MESO-14 cells (142.51±26.85, 165.16±12.91 vs. 92.31±8.69). In vivo, the tumor detection rate of fluorescence diagnosis was 1.1-4.5 times higher than that of white light. The mean number of metastases detected by the PDD was significantly higher than that of white light for FT821 (p=0.004), Ma44-3 (p=0.006) and Y-MESO-14 cell lines (p=0.005), but not for A549, NCI-H290 and MSTO-211H cell lines. Small lesions were detected by fluorescence diagnosis even though the lesions were invisible macroscopically under white light. CONCLUSION: Our results suggest the possibility of clinical application of fluorescence diagnosis with intrapleural malignant tumors.
Jun Kishi, Yasuhiko Nishioka, Tomomi Kuwahara, Souji Kakiuchi, Momoyo Azuma, Yoshinori Aono, Hideki Makino, Katsuhiro Kinoshita, Masami Kishi, R Batmunkh, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Blockade of Th1 chemokine receptors ameliorates pulmonary granulomatosis in mice., The European Respiratory Journal, Vol.38, No.2, 415-424, 2011.
(要約)
Sarcoidosis is a granulomatous disease of unknown aetiology. We identified immunological targets for the treatment of pulmonary granulomatosis using a murine model generated with Propionibacterium acnes. Sensitisation and challenge using heat-killed P. acnes and dendritic cells (DCs) were performed to produce pulmonary granulomatosis in C57BL/6 mice. Immunological analyses using ELISA as well as cDNA microarray analysis were used to search for cytokines or chemokines associated with the formation of granulomas in the lungs. Co-administration of P. acnes and DCs reproducibly induced the formation of pulmonary granulomas, which resembled sarcoid granulomas. The cDNA microarray assay demonstrated that the gene expression of CXCL9 and CXCL10, ligands for CXCR3, and of CCL4, a ligand for CCR5, was strongly upregulated during granulomatosis. ELISA confirmed that levels of CXCL9 and CXCL10 as well as T-helper (Th)1 cytokines and chemokines including tumour necrosis factor-α and interferon-γ were elevated in bronchoalveolar lavage fluid (BALF). The blockade of Th1 chemokine receptors using TAK-779, a dual blocker for CXCR3 and CCR5, led to reduced numbers of CXCR3+CD4+ and CCR5+CD4+ T-cells in BALF. Furthermore, administration of TAK-779 ameliorated the granulomatosis. The targeted inhibition of Th1 chemokines might be useful for inhibiting Th1-biased granulomatous diseases, including sarcoidosis.
Malignant pleural mesothelioma (MPM), arises from the mesothelial cells, is difficult to be diagnosed at an early stage, and is refractory to conventional chemotherapy and radiotherapy. Therefore, the establishment of novel effective therapies is necessary to improve the prognosis for many patients with this disease. Recent studies have demonstrated that angiogenesis plays a significant role in MPM progression, suggesting the importance of tumor vessels as therapeutic targets. To explore molecular pathogenesis and evaluate the efficacy of vascular targeting therapy in MPM, we developed orthotopic implantation SCID mouse models of MPM. We found that selective VEGF inhibitors were effective only in the treatment of high-VEGF-producing MPM models. On the other hand, multiple kinase inhibitor E7080, with inhibitory activity against various angiogenic cytokine receptors, suppressed the progression and prolonged survival of both high-VEGF-producing and low-VEGF-producing MPM models. Further understanding of the functional characteristics of tumor angiogenesis may be essential to improve targeting therapies in MPM. In this review, we introduce current status of clinical strategies and novel therapeutic approaches against angiogenesis in MPM.
Yasuhiko Nishioka, Yoshinori Aono and Saburo Sone : Role of tyrosine kinase inhibitors in tumor immunology., Immunotherapy, Vol.3, No.1, 107-116, 2011.
(要約)
Various immune cells are involved in both innate and acquired immunity against tumors. NK cells and cytotoxic T lymphocytes play a role as effector cells to directly kill tumor cells. On the other hand, antigen-presenting cells, particularly dendritic cells, control tumor-specific immune responses. In addition, much focus has been paid on the immune regulatory cells in tumor sites, including CD4(+)CD25(+) regulatory T cells and myeloid-derived suppressor cells. The recent advances in molecular-targeted therapy for cancer have provided small-molecule kinase inhibitors, which are effective for several hematopoietic malignancies as well as solid tumors in the clinical setting. Most drugs generally have inhibitory effects on several kinases, including tyrosine kinases, which are critical molecules for the survival, proliferation, migration and invasion of tumor cells. Since the host immune surveillance against tumors affects tumor progression, it is of interest to understand how these molecular-targeted drugs affect immune function in the tumor-bearing host. Besides this, there are emerging findings that myeloid cells could be involved in tumor angiogenesis. In this article, we address the role of tyrosine kinase inhibitors in tumor immunology by summarizing their effects on myeloid cells, such as antigen-presenting cells and regulatory cells, and their role in tumor immunity and angiogenesis.
Takanori Kanematsu, Masaki Hanibuchi, Hideki Tomimoto, Shoji Sakiyama, Koichiro Kenzaki, Kazuya Kondo, Bando Hiroyasu, Haku Takashi, Yoneda Kazuo, Hirose Toshiyuki, Toyoda Yuko, Hisatsugu Goto, Sakaguchi Satoshi, Katsuhiro Kinoshita, Momoyo Azuma, Kakiuchi Soji, Jun Kishi, Masahiko Azuma, Tada Hiroya, Sumitomo Masayuki, Yasuhiko Nishioka, Seiji Yano and Saburo Sone : Epidemiological and clinical features of lung cancer patients from 1999 to 2009 in Tokushima Prefecture of Japan, The Journal of Medical Investigation : JMI, Vol.57, No.3,4, 326-333, 2010.
(要約)
Lung cancer is the leading cause of malignancy-related death worldwide. In the present study, we reviewed the epidemiologic and clinical features of lung cancer in Tokushima Prefecture, Japan. Between January 1999 and December 2009, 2,183 patients with lung cancer were enrolled in this study. One thousand five hundred ninety-one (73%) patients were male and 592 (27%) patients were female. Median age was 70 years, with a range of 15-93 years. Seventy-six percent of patients had smoking history. One thousand nine hundred five (87%) patients were non-small cell lung cancer and the predominant histological type was adenocarcinoma (51%). Among all 2,183 patients, 702 (32%) belonged to elderly population. Four hundred seventy-one (22%), 213 (10%), 24 (1%), 116 (5%), 238 (11%), 370 (17%) and 678 (31%) patients had stage IA, IB, IIA, IIB, IIIA, IIIB and IV lung cancer, respectively. In Tokushima University Hospital, 516 (29%), 191 (11%), 58 (3%), 755 (43%) and 216 (12%) patients were initially treated with chemotherapy, chemo-radiotherapy, thoracic radiotherapy, operation and best supportive care, respectively. The median time to progression (TTP) and the median survival time (MST) of patients treated with chemotherapy and chemo-radiotherapy were 3.5 months, 13.0 months and 7.0 months, 18.0 months, respectively. The median TTP and the MST of 33 elderly patients treated with chemotherapy were 3.3 months and 18.0 months, respectively, which were comparable with those of total population. These results indicated the benefit of chemotherapy in elderly patients with advanced lung cancer by proper selection.
(キーワード)
Adolescent / Adult / Age Factors / Aged / Aged, 80 and over / Carcinoma, Non-Small-Cell Lung / Carcinoma, Small Cell / Female / Humans / Japan / Kaplan-Meier Estimate / Lung Neoplasms / Male / Middle Aged / Risk Factors / 喫煙 (smoking) / Young Adult
Hirohisa Ogawa, Masahiko Azuma, S Muto, Yasuhiko Nishioka, A Honjo, T Tezuka, Hisanori Uehara, Keisuke Izumi, A Itai and Saburo Sone : IκB kinase β inhibitor IMD-0354 suppresses airway remodelling in a Dermatophagoides pteronyssinus-sensitized mouse model of chronic asthma, Clinical and Experimental Allergy, Vol.41, No.1, 104-115, 2010.
(要約)
Nuclear factor (NF)-κB is a transcription factor that regulates cytokine and chemokine production in various inflammatory diseases, including bronchial asthma. IκB kinase (IKK) β is important for NF-κB activation in inflammatory conditions, and is possibly related to airway remodelling. Thus, inhibition of the IKKβ-NF-κB pathway may be an ideal strategy for the management of airway remodelling. We examined the effects of a newly synthesized IKKβ inhibitor, IMD-0354, in a chronic allergen exposure model of bronchial asthma in mice. A chronic mouse model was generated by challenge with house dust mite antigen (Dermatophagoides pteronyssinus). IMD-0354 was administrated intraperitoneally in therapeutic groups. Lung histopathology, hyperresponsiveness and the concentrations of mediators and molecules in supernatants of lung homogenates were determined. NF-κB activation was inhibited by prolonged periods of IMD-0354 administration. IMD-0354 reduced the numbers of bronchial eosinophils. IMD-0354 also inhibited the pathological features of airway remodelling, including goblet cell hyperplasia, subepithelial fibrosis, collagen deposition and smooth muscle hypertrophy. Inhibition of these structural changes by IMD-0354 was the result of the suppressing the production and activation of remodelling-related mediators, such as TGF-β, via inhibition of IKKβ. IMD-0354 inhibited IL-13 and IL-1β production, and it restored the production of IFN-γ. It also ameliorated airway hyperresponsiveness. IKKβ plays crucial roles in airway inflammation and remodelling in a chronic mouse model of asthma. A specific IKKβ inhibitor, IMD-0354, may be therapeutically beneficial for treating airway inflammation and remodelling in chronic asthma.
Satoshi Sakaguchi, Hisatsugu Goto, Masaki Hanibuchi, Shinsaku Otsuka, Hirokazu Ogino, Souji Kakiuchi, Hisanori Uehara, Seiji Yano, Yasuhiko Nishioka and Saburo Sone : Gender difference in bone metastasis of human small cell lung cancer, SBC-5 cells in natural killer-cell depleted severe combined immunodeficient mice., Clinical & Experimental Metastasis, Vol.27, No.5, 351-359, 2010.
(要約)
Lung cancer frequently develops multiple organ metastases, which thus makes this disease a leading cause of malignancy-related death worldwide. A gender difference is reported to affect the incidence and mortality of lung cancer; however, whether and how the gender difference is involved in lung cancer metastasis is unclear. This study evaluated the gender difference in multiple organ metastases in human small cell lung cancer (SBC-5) cells by using natural killer cell-depleted severe combined immunodeficient mice. Among multiple organ metastases, only bone metastasis formation significantly increased in female mice in comparison to males, while no significant difference was observed in the metastases to the liver and lungs. The suppression of androgen by castration or androgen receptor antagonist treatment in male mice also induced a significant increase of bone metastases. The number of osteoclasts in the bone metastatic lesions was greater in female mice and in mice with androgen suppression than in control male. However, there was no significant difference in the serum concentration of parathyroid hormone-related protein (PTHrP) associated with gender or androgen suppression. An in vitro study also indicated that sex steroid treatment had no effect on the proliferation or PTHrP production in SBC-5 cells. These results indicate that the balance of sex steroids therefore plays an important role in the formation of bone metastasis in small cell lung cancer, and suggests diverse mechanisms of interaction between cancer cells and host cells in the bone microenvironment.
(キーワード)
Androgens / Animals / Base Sequence / Bone Neoplasms / Carcinoma, Small Cell / Cell Line, Tumor / Cell Proliferation / DNA Primers / 女性 (female) / Humans / Killer Cells, Natural / Lung Neoplasms / Male / Mice / Mice, SCID / Orchiectomy / Parathyroid Hormone-Related Protein / Reverse Transcriptase Polymerase Chain Reaction / Sex Factors
Tadaaki Yamada, Kunio Matsumoto, Wei Wang, Qi Li, Yasuhiko Nishioka, Yoshitaka Sekido, Saburo Sone and Seiji Yano : Hepatocyte growth factor reduces susceptibility to an irreversible epidermal growth factor receptor inhibitor in EGFR-T790M mutant lung cancer., Clinical Cancer Research, Vol.16, No.1, 174-183, 2010.
(要約)
The secondary T790M mutation in epidermal growth factor receptor (EGFR) is the most frequent cause of acquired resistance to the reversible EGFR tyrosine kinase inhibitors (EGFR-TKI), gefitinib and erlotinib, in lung cancer. Irreversible EGFR-TKIs are expected to overcome the reversible EGFR-TKI resistance of lung cancer harboring T790M mutation in EGFR. However, it is clear that resistance may also develop to this class of inhibitors. We showed previously that hepatocyte growth factor (HGF) induced gefitinib resistance of lung cancer harboring EGFR-activating mutations. Here, we investigated whether HGF induced resistance to the irreversible EGFR-TKI, CL-387,785, in lung cancer cells (H1975) harboring both L858R activating mutation and T790M secondary mutation in EGFR.
Tohru Sakai, Sakina Furoku, Mariko Nakamoto, Emi Shuto, Toshio Hosaka, Yasuhiko Nishioka and Saburo Sone : The Soy isoflavone equol enhances antigen-specific IgE production in ovalbumin-immunized BALB/c mice., Journal of Nutritional Science and Vitaminology, Vol.56, No.1, 72-76, 2010.
(要約)
Although an immunomodulatory role of the soy isoflavone genistein has been demonstrated, the effects of other soy isoflavones on induction of antigen (Ag)-specific immune responses are not known. In this study, we therefore investigated the effects of daidzein and equol on ovalbumin (OVA)-specific T cell and B cell responses in BALB/c mice. Mice that had been treated with 20 mg/kg equol showed a significantly higher level of OVA-specific IgE than control mice. Levels of interferon (IFN)-gamma and interleukin (IL)-4 production were not different between the control and equol groups. However, IL-13 production level in mice administered 20 mg/kg equol was significantly higher than that in control mice. Strong induction of OVA-specific IgE production by equol was also observed in ovariectomized BALB/c mice, suggesting that the immunomodulatory effect of equol is not affected by endogenous estrogen.
Battur Lkhagvaa, Kenji Tani, Keiko Sato, Yuko Toyoda, Chiyuki Suzuka and Saburo Sone : Bestatin, an inhibitor for aminopeptidases, modulates the production of cytokines and chemokines by activated monocytes and macrophages., Cytokine, Vol.44, No.3, 386-391, 2008.
(要約)
The aim of this study was to clarify the effect of bestatin, an aminopeptidase inhibitor, on the production of cytokines from peripheral blood monocytes and alveolar macrophages (AM). Human monocytes isolated from peripheral blood of healthy volunteers were incubated with or without lipopolysaccharide (LPS) in the presence or absence of bestatin. AM obtained from patients with sarcoidosis were incubated in the presence or absence of bestatin. The concentration of cytokines in the culture supernatant was determined by enzyme-linked immunosorbent assay. The expression of mRNA was determined by reverse transcription polymerase chain reaction. Bestatin suppressed the production and expression of proinflammatory cytokines and chemokines, interleukin (IL)-6, CXCL8/IL-8, CCL3/macrophage inflammatory protein (MIP)-1alpha by LPS-stimulated monocytes. The mean percentage of the inhibition of IL-6, CXCL8/IL-8, CCL3/MIP-1alpha by bestatin at a concentration of 50 microg/mL was 71.2%, 29.7% and 61.0%, respectively. On the other hand, bestatin increased the production and mRNA expression of IL-10 by LPS-stimulated monocytes. The treatment with bestatin significantly inhibited the production of IL-6 and CXCL8/IL-8 by AM from patients with sarcoidosis. The data presented here indicate that bestatin suppresses the production of the pro-inflammatory cytokines and stimulates the anti-inflammatory cytokine by activated human monocytes. This study suggests that bestatin may be useful as an anti-inflammatory agent in various inflammatory diseases.
Akemi Sugita, Hirohisa Ogawa, Masahiko Azuma, Susumu Muto, Akifumi Honjo, Hiroaki Yanagawa, Yasuhiko Nishioka, Kenji Tani, Akiko Itai and Saburo Sone : Antiallergic and anti-inflammatory effects of a novel IκB kinase β inhibitor, IMD-0354, in a mouse model of allergic inflammation., International Archives of Allergy and Immunology, Vol.148, No.3, 186-198, 2008.
(要約)
BACKGROUND: Nuclear factor (NF)-kappaB is a transcription factor known to regulate allergy-associated cytokine and chemokine production related to the induction of inflammation. I kappaB kinase beta (IKK beta), which is responsible for activation of the NF-kappaB pathway, may be an ideal molecular target to inhibit this process. IMD-0354 [N-(3,5-bis-trifluoromethyl-phenyl)-5-chloro-2-hydroxy-benzamide] is an attractive novel IKK beta inhibitor that prevents the production of inflammatory cytokines in various diseases, although it is not known if IMD-0354 is effective against allergic inflammation. This study aimed to elucidate the antiallergic effects of a newly synthesized IKK beta inhibitor, IMD-0354, in a mouse model of allergic inflammation. METHODS: We generated ovalbumin (OVA)-sensitized mice which were then challenged with OVA. IMD-0354 was administered intraperitoneally to therapeutic groups. Lung histopathology and the concentrations of cytokines and chemokines in bronchoalveolar lavage fluid (BALF) and supernatants of lung homogenates were determined. RESULTS: Administration of IMD-0354 ameliorated airway hyperresponsiveness and reduced the numbers of bronchial eosinophils and mucus-producing cells in OVA-sensitized mice. The total numbers of cells and eosinophils in BALF were also reduced by treatment with IMD-0354. Treatment with IMD-0354 inhibited the production of Th2 cytokines such as interleukin (IL)-5 and IL-13 and eotaxin in the airways and/or lungs of OVA-sensitized mice, but it did not affect the restoration of Th1 cytokines such as IL-12 and interferon-gamma under the same experimental conditions. IgE production was also inhibited by IMD-0354. CONCLUSION: A specific IKK beta inhibitor, IMD-0354, improved allergic airway inflammation and hyperresponsiveness in mice. IMD-0354 may have therapeutic potential for bronchial asthma.
Reiko Tomioka, Kenji Tani, Keiko Sato, Chiyuki Suzuka, Yuko Toyoda, Jun Kishi and Saburo Sone : Observations on the occurrence of exacerbations in clinical course of systemic lupus erythematosus, The Journal of Medical Investigation : JMI, Vol.55, No.1,2, 112-119, 2008.
(要約)
Systemic lupus erythematosus (SLE) is a chronic disease that is characterized by an undulating course of exacerbations and remissions, and a major determinant of long-term prognosis is organ damage consequent to tissue injury that accompanies disease activity and toxicity of therapy. In this study, we evaluated which patients with SLE will develop an exacerbation and whether factors can be identified to predict the development of an exacerbation. Fifty-seven SLE patients (52 females) were included in this study. The exacerbation of SLE was found in 15 patients (26.3%). A relatively increased incidence of an exacerbation was found in younger SLE patients. An increased percentage of patients who had lupus nephritis at the time of diagnosis of SLE was found in patients with a subsequent exacerbation when compared with that in those without it. Increased incidence of an exacerbation was observed in patients who had decreased number of WBC and platelets, decreased level of C3 and CH50, and the presence positivity of anti-Sm antibodies at the time of the diagnosis. This study suggests that age, renal involvement, and the presence of decreased number of WBC and platelets, decreased level of complements anti-Sm antibodies are predictors of exacerbation.
M. Azuma, Yasuhiko Nishioka, Y. Aono, M. Inayama, H. Makino, Jun Kishi, Masayuki Shono, Katsuhiro Kinoshita, H. Uehara, H. ogushi, K. Izumi and Saburo Sone : Role of 1-acid glycoprorein in theraqeutic antifibrotic effects of imatinib plus macrolides in mice., American Journal of Respiratory and Critical Care Medicine, Vol.176, No.12, 1243-1250, 2007.
(要約)
Imatinib is an inhibitor of platelet-derived growth factor receptors. We have reported that treatment with imatinib inhibited bleomycin-induced pulmonary fibrosis in mice. However, late treatment with imatinib had no effect. To clarify why imatinib had no antifibrotic effect when its administration was delayed, we focused on alpha(1)-acid glycoprotein (AGP), because it was reported to bind imatinib and mediate drug resistance. The concentration of AGP in serum of mice and patients with idiopathic pulmonary fibrosis was measured by radial immunodiffusion testing. The effects of AGP in vitro were evaluated by assaying the growth of lung fibroblasts. We examined the combined effects of erythromycin (EM) or clarithromycin (CAM) on bleomycin-induced pulmonary fibrosis in mice. Addition of AGP abrogated imatinib-mediated inhibition of the growth of fibroblasts. However, treatment with EM or CAM restored the growth-inhibitory effects of imatinib. The elevated level of AGP was detected in serum and lung homogenates in bleomycin-exposed mice and reached a plateau on Day 14. Imatinib alone did not ameliorate pulmonary fibrosis when treatment was started on Day 15, whereas coadministration of imatinib and EM or CAM significantly reduced the fibrogenesis via inhibition of the growth of fibroblasts in vivo. Serum levels of AGP were higher in patients with idiopathic pulmonary fibrosis than in healthy subjects. AGP is an important regulatory factor modulating the ability of imatinib to prevent pulmonary fibrosis in mice, and combined therapy with imatinib and EM or CAM might be useful for treatment of pulmonary fibrosis.
Ali Nermin, Masanori Yoshizumi, Seiji Yano, Saburo Sone, Hideki Ohnishi, Keisuke Ishizawa, Yasuhisa Kanematsu, Koichiro Tsuchiya and Toshiaki Tamaki : The novel Src kinase inhibitor M475271 inhibits VEGF-induced vascular endothelial-cadherin and β-catenin phosphorylation but increases their association, Journal of Pharmacological Sciences, Vol.102, No.1, 112-120, 2006.
(要約)
M475271, 4-quinazolinamine, N-(2-chloro-5-methoxyphenyl)-6-methoxy-7-[(1-methyl-4-piperidinyl) methoxy]-(9Cl), is a new anilinoquinazoline derivative that displays selective inhibition of Src kinase activity and tumor growth in vivo. Vascular endothelial growth factor (VEGF)-induced angiogenesis plays a pivotal role in tumor growth and metastasis. Vascular endothelial (VE)-cadherin is an endothelial cell-specific adhesion molecule that can interact with the cytoskeleton via several anchoring molecules such as beta-catenin. Here, we examined the effect of M475271 on VE-cadherin and beta-catenin phosphorylation and association. We also examined its effect on VEGF-induced human umbilical vein endothelial cell (HUVEC) proliferation, migration, and tube formation. The findings reveal pretreatment with M475271 significantly inhibits VEGF-induced VE-cadherin and beta-catenin phosphorylation. However, M475271 significantly increases VE-cadherin and beta-catenin association compared to the VEGF-treated group. Confocal laser microscopic examination confirmed the augmentation effect of M475271 on VE-cadherin and beta-catenin association. Finally, M475271 was shown to have inhibitory effects comparable to those of PP2 and Herbimycin A on VEGF-induced HUVEC proliferation, migration, and tube formation. These findings suggest that M475271 attenuates VEGF-induced angiogenesis by maintaining cell-cell junction stability. Although the involvement of other signaling molecules cannot be ruled out, M475271 has potential as a drug for the inhibition of the angiogenesis needed for tumor growth and metastasis.
Seiji Yano, Hiroaki Muguruma, Yuka Matsumori, Hisatsugu Goto, Emiko Nakataki, Nobutaka Edakuni, Hideki Tomimoto, Soji Kakiuchi, Akihiko Yamamoto, Hisanori Uehara, Anderson Ryan and Saburo Sone : Antitumor Vascular Strategy for Controlling Experimental Metastatic Spread of Human Small-Cell Lung Cancer Cells with ZD6474 in Natural Killer Cell Depleted Severe Combined Immunodeficient Mice, Clinical Cancer Research, Vol.11, No.24, 8789-8798, 2005.
(要約)
Small-cell lung cancer is often characterized by rapid growth and metastatic spread. Because tumor growth and metastasis are angiogenesis dependent, there is great interest in therapeutic strategies that aim to inhibit tumor angiogenesis. The effect of ZD6474, an orally available inhibitor of vascular endothelial growth factor receptor-2 (VEGFR-2) and epidermal growth factor tyrosine kinases, was studied in experimental multiple-organ metastasis models with human small-cell lung cancer cell lines (SBC-3 or SBC-5) in natural killer cell-depleted severe combined immunodeficient mice. Intravenously inoculated SBC-5 cells produced experimental metastases in the liver, lung, and bone whereas SBC-3 cells produced the metastases in the liver, systemic lymph nodes, and kidneys. Daily oral treatment with ZD6474 (50 mg/kg), started on day 14 (after the establishment of micrometastases), significantly reduced the frequency of large (>3 mm) metastatic colonies (in the liver and lymph nodes) and osteolytic bone lesions. ZD6474 treatment did not significantly reduce the frequency of small (<2-3 mm) metastatic lesions found in the lung (SBC-5) or kidney (SBC-3), consistent with an antiangiogenic mechanism of action. Immunohistochemical analysis of SBC-5 metastatic deposits in the liver showed that ZD6474 treatment inhibited VEGFR-2 activation and induced apoptosis of tumor-associated endothelial cells, resulting in decreasing tumor microvessel density. ZD6474 treatment was also associated with a decrease in tumor cell proliferation and an increase in tumor cell apoptosis. The antitumor effects of ZD6474 were considered likely to be due to inhibition of VEGFR-2 tyrosine kinase because gefitinib, a small-molecule inhibitor of epidermal growth factor receptor tyrosine kinase, was inactive in these models. These results suggest that ZD6474 may be of potential therapeutic value in inhibiting the growth of metastatic small-cell lung cancer in humans. Phase II trials with ZD6474 are currently ongoing in a range of solid tumors.
Chiyuki Furukawa, Yataro Daigo, Nobuhisa Ishikawa, Tatsuya Kato, Tomoo Ito, Eiju Tsuchiya, Saburo Sone and Yusuke Nakamura : Plakophilin 3 Oncogene as Prognostic Marker and Therapeutic Target for Lung Cancer, Cancer Research, Vol.65, No.16, 7102-7110, 2005.
(要約)
We investigated gene expression profiles of non-small cell lung carcinomas (NSCLC) to screen candidate molecules that might be useful as diagnostic markers or for development of novel molecular-targeting therapies. Here we report evidence that a member of the armadillo protein family, plakophilin 3 (PKP3), is a potential molecular target for treatment of lung cancers and might also serve as a prognostic indicator. We documented elevated expression of PKP3 in the great majority of NSCLC samples examined. Treatment of NSCLC cells with small interfering RNAs of PKP3 suppressed growth of the cancer cells; on the other hand, induction of exogenous expression of PKP3 conferred growth-promoting activity on COS-7 cells and enhanced their mobility in vitro. To investigate its function, we searched for PKP3-interacting proteins and identified dynamin 1-like, which was also activated in NSCLC. In addition, a high level of PKP3 expression was associated with poor survival as well as disease stage and node status for patients with lung adenocarcinoma, suggesting an important role of the protein in development and progression of this disease. As our data imply that up-regulation of PKP3 is a frequent and important feature of lung carcinogenesis, we suggest that targeting the PKP3 molecule might hold promise for development of a new therapeutic and diagnostic strategy for clinical management of lung cancers.
Ali Nermin, Masanori Yoshizumi, Yoshiko Fujita, Yuki Izawa, Yasuhisa Kanematsu, Keisuke Ishizawa, Koichiro Tsuchiya, Seiji Yano, Saburo Sone and Toshiaki Tamaki : A Novel Src Kinase Inhibitor, M475271, Inhibits VEGF-Induced Human Umbilical Vein Endothelial Cell Proliferation and Migration, Journal of Pharmacological Sciences, Vol.98, No.2, 130-141, 2005.
(要約)
Vascular endothelial growth factor (VEGF) was reported to be a potent proangiogenic factor that plays a pivotal role in both physiological and pathological angiogenesis. M475271, 4-quinazolinamine, N-(2-chloro-5-methoxyphenyl)-6-methoxy-7-[(1-methyl-4-piperidinyl) methoxy]-(9Cl), is a new anilinoquinazoline derivative that showed selective inhibition of Src kinase activity and tumor growth in vivo. Here, we examined the effect of M475271 on VEGF-induced human umbilical vein endothelial cell (HUVEC) proliferation and migration and their intracellular mechanisms. Our findings showed that M475271 pretreatment resulted in a significant inhibition of VEGF-induced HUVEC proliferation, [(3)H]thymidine incorporation, and migration. M475271 inhibited VEGF-induced Flk-1 and Src phosphorylation and their association. Confocal laser microscopic examination confirmed the inhibitory effect of M475271 on VEGF-induced Flk-1/Src association. M475271 inhibited VEGF-induced extracellular signal-regulated kinase1/2 (ERK1/2) and p38 but not Akt activation in a concentration-dependent manner. M475271, PI3-K inhibitor, and p38 inhibitor inhibited VEGF-induced HUVEC proliferation and migration. However, a MEK1/2 inhibitor inhibited VEGF-induced proliferation but not migration. These findings suggest that M475271 attenuates VEGF-induced HUVEC proliferation and migration through the inhibition of signaling pathways involving Src, ERK1/2, and/or p38. Taken together, these data indicate that M475271 may be a useful candidate for inhibition of endothelial cell proliferation and migration relevant to angiogenesis.
(キーワード)
M475271 / vascular endothelial growth factor / human umbilical vein endothelial cell proliferation and migration / angiogenesis
Yoshinori Aono, Yasuhiko Nishioka, Mami Inayama, Jun Kishi, Momoyo Ugai, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Imatinib as a Novel Antifibrotic Agent in Bleomycin-induced Pulmonary Fibrosis in Mice, American Journal of Respiratory and Critical Care Medicine, Vol.171, No.11, 1279-1285, 2005.
(要約)
Imatinib mesylate is a potent and specific tyrosine kinase inhibitor against c-ABL, BCR-ABL, and c-KIT, and has been demonstrated to be highly active in chronic myeloid leukemia and gastrointestinal stromal tumors. We examined the antifibrotic effects of imatinib using a bleomycin-induced lung fibrosis model in mice because imatinib also inhibits tyrosine kinase of platelet-derived growth factor receptors (PDGFRs). Imatinib inhibited the growth of primary murine lung fibroblasts and the autophosphorylation of PDGFR-beta induced by PDGF. Administration of imatinib significantly prevented bleomycin-induced pulmonary fibrosis in mice, partly by reducing the number of mesenchymal cells incorporating bromodeoxyuridine. Analysis of bronchoalveolar lavage cells demonstrated that imatinib did not suppress early inflammation on Days 7 and 14 caused by bleomycin. These results suggest that imatinib has the potential to prevent pulmonary fibrosis by inhibiting the proliferation of mesenchymal cells, and that imatinib might be useful for the treatment of pulmonary fibrosis in humans.
Rui Zheng, Seiji Yano, Yuka Matsumori, Emiko Nakataki, Hiroaki Muguruma, Masanori Yoshizumi and Saburo Sone : SRC Tyrosine Kinase Inhibitor, M475271, Suppresses Subcutaneous Growth and Production of Lung Metastasis Via Inhibition of Proliferation, Invasion, and Vascularization of Human Lung Adenocarcinoma Cells, Clinical & Experimental Metastasis, Vol.22, No.3, 195-204, 2005.
(要約)
Src, a proto-oncogene, has been strongly implicated in the growth, progression and metastasis of a number of human cancers. Its role in lung cancer is, however, still unknown. In the present study, we assessed the expression of Src in three different human lung adenocarcinoma cell lines (PC-9, PC14PE6, A549), and explored the effect of a novel Src kinase inhibitor, M475271, on the behavior of the cell lines. The three cell lines expressed various levels of auto-phosphorylated Src. While M475271 reduced Src-phosphorylation and invasiveness of all three cell lines, it inhibited the proliferation of PC-9 and A549 cells with highly phosphorylated Src, but not PC14PE6 cells. We further examined the effect of M475271 on subcutaneous tumors and lung metastasis caused by PC-9 and/or A549 cells in NK-cell depleted SCID mice. Daily oral treatment with M475271 inhibited the growth of subcutaneous tumors with PC-9 and A549 cells via inhibition of tumor cells proliferation, VEGF production and/or vascularization in the mice in a dose-dependent manner. In the metastasis model with A549 cells, the lung weight in the M475271 (50 mg/kg)-treated group was less than that of the control group, despite no difference in the number of metastatic nodules. Our results suggest that inhibition of tyrosine kinase Src by M475271 could reduce the growth, invasion and VEGF-mediated neovascularization of lung adenocarcinoma cells, resulting in inhibition of growth of subcutaneous tumors and lung metastasis. Therefore, a novel Src tyrosine kinase inhibitor, M475271, might be helpful for controlling the progression of human lung adenocarcinoma.
Yanjmaa Bira, Kenji Tani, Yasuhiko Nishioka, juuya Miyata, Keiko Sato, Akihito Hayashi, Yutaka Nakaya and Saburo Sone : Transforming growth factor β stimulates rheumatoid synovial fibroblasts via the type receptor, Modern Rheumatology, Vol.15, No.2, 108-113, 2005.
(要約)
Transforming growth factor (TGF)-beta regulates the function of fibroblasts, and has been shown to have a role in the pathogenesis of rheumatoid arthritis (RA) because several studies have demonstrated the presence of TGF-beta in the synovial tissue and synovial fluids of RA patients. In this study, we examined the expression of TGF-beta receptors in synovial fibroblasts of patients with RA and demonstrated the significance in functional responses of synovial fibroblasts to TGF-beta in this disorder. Transforming growth factor beta1 stimulated the expression of connective tissue growth factor (CTGF) in fibroblasts of patients with RA more than in those of patients with osteoarthritis (OA). Transforming growth factor beta1 induced the chemotactic migration of RA synovial fibroblasts and inhibited their proliferation significantly more than OA synovial fibroblasts. Both RA and OA synovial fibroblasts expressed detectable amounts of TGF-beta receptor type II mRNA, but the expression was higher in RA patients than in OA patients, as assessed by reverse transcriptase-polymerase chain reaction. There was no significant difference in the expression of TGF-beta receptor type I or type III in synovial fibroblasts between RA and OA patients. These results indicate that synovial fibroblasts of RA patients express the increased TGF-beta receptor type II, which is associated with altered responses to TGF-beta observed in CTGF expression, chemotaxis, and proliferation of RA synovial fibroblasts, and may have an important role in the pathogenesis of RA.
Hiroki Kuniyasu, Seiji Yano, Takamitsu Sasaki, Tomonori Sasahira, Saburo Sone and Hitoshi Ohmori : Colon Cancer Cell-Derived High Mobility Group 1/Amphoterin Induces Growth Inhibition and Apoptosis in Macrophages, The American Journal of Pathology, Vol.166, No.3, 751-760, 2005.
(要約)
High mobility group (HMGB)1/amphoterin is a multifunctional cytokine involved in invasion and metastasis of cancer and in inflammation. To investigate HMGB1/amphoterin effects on macrophages, U937 human monocytic leukemia cells and rat peritoneal and human alveolar macrophages were examined. U937 cells expressed low levels of an HMGB1/amphoterin receptor, receptor for advanced glycation end-products (RAGE), whereas RAGE production was induced in differentiated phorbol 12-myristate 13-acetate (PMA)-U937 cells. Treatment with cultured medium of HMGB1/amphoterin-secreting WiDr human colon cancer cells showed growth inhibition of both U937 and PMA-U937 cells and apoptosis in PMA-U937 cells. The number of PMA-U937 cells was markedly decreased by co-culture with WiDr cells exposed to HMGB1/amphoterin sense S-oligodeoxynucleotide (ODN) in spheroids or monolayers. In contrast, PMA-U937 cells co-cultured with WiDr cells exposed to HMGB1/amphoterin anti-sense S-ODN were preserved in number. PMA-U937 cells exposed to RAGE anti-sense S-ODN were insensitive to WiDr-cultured medium. Recombinant human HMGB1/amphoterin induced growth inhibition in thioglycollate-induced rat peritoneal macrophages, PMA-U937 cells, and human alveolar macrophages, an effect that was abrogated by absorption with anti-HMGB1 antibody. Phosphorylation of JNK and Rac1 was induced in PMA-U937 cells treated with HMGB1/amphoterin. These results suggest that HMGB1/amphoterin induces growth inhibition and apoptosis in macrophages through RAGE intracellular signaling pathway.
(キーワード)
Animals / Apoptosis / Cell Line / Cell Line, Tumor / Cell Proliferation / Coculture Techniques / Colonic Neoplasms / Culture Media, Conditioned / Glycosylation End Products, Advanced / HMGB1 Protein / Humans / Immunoblotting / MAP Kinase Signaling System / Macrophages / Nitric Oxide Synthase / Nitric Oxide Synthase Type II / Nitrites / Oligonucleotides / Oligonucleotides, Antisense / Rats / Rats, Inbred F344 / Recombinant Proteins / Signal Transduction / Tetradecanoylphorbol Acetate / Time Factors / U937 Cells
Masaki Hanibuchi, Yuka Matsumori, Yasuhiko Nishioka and Saburo Sone : A case of sarcoidosis accompanying squamous cell carcinoma in the mandibular gingiva, The Journal of Medical Investigation : JMI, Vol.52, No.1-2, 118-121, 2005.
(要約)
A 51-year-old man with a history of gingival cancer two years previously was referred to our hospital for further examination of chest abnormal shadow. Bilateral hilar and mediastinal lymphadenopathy, diffuse small nodular opacities and pleural nodules were observed in chest high resolution CT. Serum angiotensin converting enzyme and lysozyme were elevated. Transbronchial lung biopsy specimens demonstrated non-caseous granuloma. CD4-positive lymphocytes were increased in broncho-alveolar lavage (CD4/CD8 ratio 5.47). The patient was diagnosed as having sarcoidosis. Radiological findings were improved and serum angiotensin converting enzyme level was decreased to within the normal range by corticosteroid therapy (prednisolone 30 mg/day). This is the first report of sarcoidosis accompanying the gingival cancer.
Kazuyoshi Manabe, Yasuhiko Nishioka, Jun Kishi, Mami Inayama, Yoshinori Aono, Yoichi Nakamura, Fumitaka Ogushi, Hiroyasu Bando, Kenji Tani and Saburo Sone : Elevation of macrophage-derived chemokine in eosinophilic pneumonia: a role of alveolar macrophages, The Journal of Medical Investigation : JMI, Vol.52, No.1,2, 85-92, 2005.
(要約)
Macrophage-derived chemokine (MDC/CCL22) and thymus-and activation-regulated chemokine (TARC/CCL17) are ligands for CC chemokine receptor 4. Recently, TARC has been reported to play a role in the pathogenesis of idiopathic eosinophilic pneumonia (IEP). The purpose of this study was to evaluate the role of MDC in IEP and other interstitial lung diseases (ILDs). MDC and TARC in the bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay in patients with ILDs and healthy volunteers (HV). We also examined the expression of MDC mRNA in alveolar macrophages (AM) by real-time quantitative reverse transcriptase-polymerase chain reaction. Both MDC and TARC were detected only in BALF obtained from IEP patients. The concentration of MDC was higher than that of TARC in all cases. The level of MDC in IEP correlated with that of TARC. AM from IEP patients expressed a significantly higher amount of MDC than that from HV at the levels of protein and mRNA. MDC in BALF from IEP dramatically decreased when patients achieved remission. These findings suggest that MDC, in addition to TARC, might be involved in the pathogenesis of IEP, and AM play a role in the elevation of MDC in IEP.
Makoto Kimura, Kenji Tani, Junya Miyata, Keiko Sato, Akihito Hayashi, Shinsaku Otsuka, Tomoyuki Urata and Saburo Sone : The significance of cathepsins, thrombin and aminopeptidase in diffuse interstitial lung diseases, The Journal of Medical Investigation : JMI, Vol.52, No.1,2, 93-100, 2005.
(要約)
To determine the significance of proteases in interstitial lung diseases, we examined the activity of cathepsins, thrombin, and aminopeptidase in bronchoalveolar lavage (BAL) fluid from patients with these disorders. Significantly increased activities of cathepsin H and aminopeptidase were detected in BAL fluid from patients with idiopathic pulmonary fibrosis (IPF), cryptogenic organizing pneumonia (COP), chronic eosinophilic pneumonia (CEP) and hypersensitivity pneumonitis (HP). Significantly higher activity of cathepsin B was found in BAL fluid from patients with CEP. The activity of thrombin was significantly higher in patients with IPF and CEP. In patients with IPF, there were significant correlations between neutrophil number and the activity of cathepsin B, cathepsin H or aminopeptidase. In patients with COP and HP, the activity of the proteases was significantly higher in patients with higher number of lymphocytes than in those with lower number of lymphocytes. The present study suggests that the activity of the proteases is a useful marker in activity of the interstitial lung diseases, and may have a role in the pathogenesis of these disorders.
(キーワード)
interstitial lung diseases / cathepsin H / cathepsin B / cathepsin G / thrombin / aminopeptidase
Seiji Yano, Emiko Nakataki, Shinsaku Ohtsuka, Mami Inayama, Hideki Tomimoto, Nobutaka Edakuni, Soji Kakiuchi, Naoki Nishikubo, Hiroaki Muguruma and Saburo Sone : Retreatment of Lung Adenocarcinoma Patients With Gefitinib Who Had Experienced Favorable Results From Their Initial Treatment With This Selective Epidermal Growth Factor Receptor Inhibitor: A Report of Three Cases, Oncology Research, Vol.15, No.2, 107-111, 2005.
(要約)
Gefitinib is a selective inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinases, and shows favorable antitumor activity against chemorefractory non-small cell lung cancer (NSCLC). The majority of responders (patients who are sensitive to gefitinib), however, relapse within 1.5 years, indicating an acquired resistance to gefitinib. Here we report three chemotherapy refractory NSCLC patients who were retreated with gefitinib. All three cases were nonsmokers and showed an adenocarcinoma histology. While they had experienced successful control from their initial treatment with gefitinib for more than 12 months, gefitinib therapy was terminated because two cases (cases 1 and 3) relapsed during the therapy and case 2 suffered alveolar hemorrhage. After more than 7 months from the time of discontinuation of the initial gefitinib treatment, they were retreated with gefitinib, as further tumor progression was observed. Of the three cases, cases 1 and 2 were well controlled by retreatment with gefitinib monotherapy for more than 7 months, suggesting sensitivity to retreatment. Case 3 also showed a regression in size of several tumors, while some other lesions progressively enlarged and developed a malignant pleural effusion after 4 months. These observations suggest the possibility that retreatment with gefitinib might be useful when 1) initial treatment shows a favorable clinical response, and 2) there has been a period of time following the termination of the initial gefitinib treatment.
Soji Kakiuchi, Yataro Daigo, Nobuhisa Ishikawa, Chiyuki Furukawa, Tatsuhiko Tsunoda, Seiji Yano, Kazuhiko Nakagawa, Takashi Tsuruo, Nobuoki Kohno, Masahiro Fukuoka, Saburo Sone and Yusuke Nakamura : Prediction of sensitivity of advanced non-small cell lung cancers to gefitinib (Iressa, ZD1839), Human Molecular Genetics, Vol.13, No.24, 3029-3043, 2004.
(要約)
Gefitinib (Iressa, ZD1839), an inhibitor of epidermal growth factor receptor-tyrosine kinase, has shown potent anti-tumor effects and improved symptoms and quality-of-life of a subset of patients with advanced non-small cell lung cancer (NSCLC). However, a large portion of the patients showed no effect to this agent. To establish a method to predict the response of NSCLC patients to gefitinib, we used a genome-wide cDNA microarray to analyze 33 biopsy samples of advanced NSCLC from patients who had been treated with an identical protocol of second to seventh line gefitinib monotherapy. We identified 51 genes whose expression differed significantly between seven responders and 10 non-responders to the drug. We selected the 12 genes that showed the most significant differences to establish a numerical scoring system (GRS, gefitinib response score), for predicting response to gefitinib treatment. The GRS system clearly separated the two groups without any overlap, and accurately predicted responses to the drug in 16 additional NSCLC cases. The system was further validated by the semi-quantitative RT-PCR, immunohistochemistry and ELISA for serological test. Moreover, we proved that the anti-apoptotic activity of amphiregulin, a protein that was significantly over-expressed in non-responders but undetectable in responders, leads to resistance of NSCLC cells to gefitinib in vitro. Our results suggested that sensitivity of a given NSCLC to gefitinib can be predicted according to expression levels of a defined set of genes that may biologically affect drug sensitivity and survival of lung cancer cells. Our scoring system might eventually lead to achievement of personalized therapy for NSCLC patients.
Hisatsugu Goto, Seiji Yano, Yuka Matsumori, Hirohisa Ogawa, David C. Blakey and Saburo Sone : Sensitization of Tumor-Associated Endothelial Cell Apoptosis by the Novel Vascular-Targeting Agent ZD6126 in Combination with Cisplatin, Clinical Cancer Research, Vol.10, No.22, 7671-7676, 2004.
(要約)
ZD6126 is a novel vascular-targeting agent that selectively disrupts the tubulin cytoskeleton of endothelial cells. In the immature vessels characteristic of tumor vasculature, this leads to endothelial cell contraction, blood vessel congestion, and, consequently, tumor cell death. ZD6126 has been shown to delay tumor growth in a range of xenograft models. The antitumor effect of ZD6126 can be increased in combination with cisplatin or radiation therapy, although the precise mechanism of this enhancement has not been demonstrated. ZD6126 treatment has also been shown to inhibit lung metastasis, and the present study has explored the potential to increase the antimetastatic effect of ZD6126 by combining with cisplatin, and the underlining mechanism has been investigated. Human lung adenocarcinoma PC14PE6 cells were injected into the tail vein of nude mice. Five weeks after injection animals were treated with ZD6126 (200 mg/kg i.p.), cisplatin (6 mg/kg i.v.), or a combination of the two agents. The animals were sacrificed 24 hours later, and the extent of lung metastases and the presence of apoptotic cells were assessed. Histologic analysis revealed that the ZD6126/cisplatin combination resulted in a 2 to 4-fold increase in the total number of tumor-associated apoptotic cells compared with either treatment alone. ZD6126 alone induced apoptosis of tumor-associated endothelial cells in tumors, and the extent of apoptosis was increased 2-fold in combination with cisplatin. The lung weight was significantly reduced, and the number of metastatic nodules significantly was lower in the combined treatment group than in the control group. These data suggest that the antimetastatic effect of the vascular-targeting agent ZD6126 can be increased by use in combination with cisplatin, which increases the incidence of endothelial cell apoptosis.
Ning Ge, Yasuhiko Nishioka, Yoichi Nakamura, Yoshio Okano, Kazuo Yoneda, Hirohisa Ogawa, Akemi Sugita, Hiroaki Yanagawa and Saburo Sone : Synthesis and Secretion of Interleukin-15 by Freshly Isolated Human Bronchial Epithelial Cells, International Archives of Allergy and Immunology, Vol.135, No.3, 235-242, 2004.
(要約)
Interleukin-15 (IL-15), which shares many functional activities of IL-2, is proposed as a potential modulator of T and natural killer (NK) cell-mediated inflammatory diseases. Since IL-15 gene is expressed in various cell types including epithelial cells, we examined how proinflammatory modulators affect IL-15 gene expression in both freshly isolated human bronchial epithelial cells (HBECs) and the human bronchial epithelial cell line BEAS-2B. HBECs were obtained from 25 patients with primary lung cancer by bronchial brushing under bronchofiberscopy. The expressions of IL-15 and its receptor were examined using reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis and enzyme-linked immunosorbent assay. IL-15 mRNA was constitutively expressed in the cells and was upregulated by several proinflammatory cytokines such as IL-1beta, tumor necrosis factor-alpha, interferon-gamma (IFN-gamma) and lipopolysaccharide. In addition, IFN-gamma but not other cytokines induced the synthesis and secretion of IL-15 protein. Investigation of IL-15 receptor expression using RT-PCR showed that IL-15Ralpha and IL-2Rbeta chains but not IL-2Ralpha or gamma chain were constitutively expressed in these cells. Bronchial epithelial cells may contribute to T and NK cell-mediated airway inflammation through IL-15 production.
Masato Okamoto, Sachiko Furuichi, Yasuhiko Nishioka, Tetsuya Oshikawa, Tomoyuki Tano, Sharif Uddin Ahmed, Kiyoshi Takeda, Shizuo Akira, Yoshiki Ryoma, Yoichiro Moriya, Motoo Saito, Saburo Sone and Mitsunobu Sato : Expression of Toll-Like Receptor 4 on Dendritic Cells Is Significant for Anticancer Effect of Dendritic Cell-Based Immunotherapy in Combination with an Active Component of OK-432, a Streptococcal Preparation, Cancer Research, Vol.64, No.15, 5461-5470, 2004.
(要約)
A lipoteichoic acid-related molecule OK-PSA is an active component of OK-432, a Streptococcus-derived anticancer immunotherapeutic agent. In the present study, we first examined the effect of OK-PSA on the maturation of dendritic cells (DCs) in vitro by using the DCs derived from 5 healthy donors and 10 patients with head and neck cancer with or without expression of toll-like receptor 4 (TLR4) or MD-2 mRNA. OK-PSA treatment effectively increased the surface expression of MHC class II, CD80, CD83, and CD86. OK-PSA-stimulated DCs secreted the cytokines that can induce helper T-cell 1 (Th1)-type T-cell response, and stimulated allogeneic T cells to produce IFN-gamma and to elicit an allogeneic antigen-specific cytotoxicity. These activities almost depended on expression of TLR4 and MD-2 genes. We next investigated the in vivo anticancer effect of intratumoral administration of syngeneic DCs followed by OK-PSA against established tumors in mice. C57BL/6 mice, which express wild-type TLR4, and C57BL/6-derived TLR4-knockout (TLR4(-/-)) mice were used. Although OK-PSA accelerated the antitumor effect of intratumoral DC administration in wild-type mice bearing syngeneic tumors, the antitumor effect of OK-PSA as well as of the combination therapy with DCs and OK-PSA was not significant in TLR4(-/-) mice. Interestingly, an administration of wild-type-mouse-derived DCs followed by OK-PSA exhibited a marked antitumor effect even in the TLR4(-/-) mice. These findings suggest that OK-PSA may be a potent adjuvant for local DC therapy, and that DC therapy followed by OK-PSA is able to elicit anticancer activity even in a TLR4-deficient host when TLR4 is expressed only in DCs injected intratumorally.
Masaki Hanibuchi, Seiji Yano, Nobutaka Edakuni, Mami Inayama and Saburo Sone : A case of early-stage lung cancer detected by autofluorescence bronchoscopy, The Journal of Medical Investigation : JMI, Vol.51, No.3-4, 234-237, 2004.
(要約)
A 71-year-old man was referred to our hospital for further examination of abnormal sputum cytology. No abnormal nodular shadows were detected in chest X-ray and chest CT. The location of the tumor was clearly identified as a defect of autofluorescence by autofluorescence bronchoscopy at the bifurcation between the left B1+2 and B3 bronchi, whereas it was quite difficult by conventional bronchoscopy. Transbronchial biopsy revealed squamous cell carcinoma. Further examinations yielded the diagnosis of early-stage lung cancer. Photodynamic therapy was performed and complete response was confirmed. This case indicates the efficacy of autofluorescence bronchoscopy for detecting early-stage lung cancer.
Tumor necrosis factor-alpha (TNF-alpha) stimulates expression of endothelial cell (EC) genes that may promote atherosclerosis in part by an activation of mitogen-activated protein (MAP) kinases. Ebselen (2-phenyl-1,2-benzisoselenazol-3[2H]-one), a selenoorganic compound, is effective for acute ischemic stroke; however, its effect on EC has not yet been elucidated. We examined the effect of ebselen on TNF-alpha-induced MAP kinase activation and adhesion molecule expression in cultured human umbilical vein endothelial cells (HUVEC). Extracellular signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK) and p38 were rapidly and significantly activated by TNF-alpha in HUVEC. TNF-alpha-induced JNK activation was inhibited by ebselen, whereas ERK1/2 and p38 were not affected. Apoptosis signal-regulated kinase 1 (ASK1) was suggested to be involved in TNF-alpha-induced JNK activation because transfection of kinase-inactive ASK1 inhibited TNF-alpha-induced JNK activation. Ebselen inhibited TNF-alpha-induced TNF receptor-associated factor 2 (TRAF2)-ASK1 complex formation and phosphorylation of stress-activated protein kinase ERK kinase 1 (SEK1), which is an upstream signaling molecule of JNK. Finally, TNF-alpha-induced activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) activation and resultant intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expressions were inhibited by ebselen. Specific inhibitors for JNK and NF-kappaB also inhibited TNF-alpha-induced ICAM-1 and VCAM-1 expressions in HUVEC. These findings suggest that ebselen prevents TNF-alpha-induced EC activation through the inhibition of TRAF2-ASK1-SEK1 signaling pathway, which leads to JNK activation. Inhibition of JNK by ebselen may imply its usefulness for the prevention of atherosclerosis relevant to EC activation.
Toyokazu Miki, Seiji Yano, Masaki Hanibuchi, Kanematsu Takanori, Hiroaki Muguruma and Saburo Sone : Parathyroid hormone-related protein (PTHRP) is responsible for production of bone metastasis, but not visceral metastasis, by human small cell lung cancer SBC-5 cells in natural killer cell-depleted SCID mice., International Journal of Cancer, Vol.108, No.4, 511-515, 2004.
(要約)
We previously established an osteolytic bone metastasis model with multiorgan dissemination in natural killer (NK) cell-depleted severe combined immunodeficient (SCID) mice using human small cell lung cancer cells (SBC-5), which highly express the parathyroid hormone-related protein (PTHrP). In our present study, we evaluated the role of PTHrP on bone metastasis by SBC-5 cells using anti-PTHrP neutralizing antibody (Ab). Anti-PTHrP Ab did not affect the proliferation or cytokine production of SBC-5 cells in vitro. Repeated intravenous injection with anti-PTHrP Ab inhibited the formation of bone metastasis in a dose-dependent manner, while the same treatment had no significant effect on the metastasis to visceral organs (lung, liver, kidney and lymph node). In addition, treatment with anti-PTHrP Ab improved the elevated serum calcium level, associated with inhibition of osteolytic bone metastasis, suggesting that anti-PTHrP Ab inhibited bone metastasis via suppression of bone resorption probably by neutralizing PTHrP. These findings suggest that PTHrP is essential for bone metastasis, but not visceral metastasis, by small cell lung cancer SBC-5 cells.
Hiroaki Yanagawa, A Sugita, M Azuma, H Ogawa, C Kitamuro, K Yoneda, K Shinkawa, Kenji Tani and Saburo Sone : Long-term Follow-up of Pulmonary Function in Bronchial Asthma Patients Treated with Pranlukast, Lung, Vol.182, No.1, 51-58, 2004.
(要約)
Clinical studies have shown that pranlukast, a selective cysteinyl leukotriene antagonist, is effective for bronchial asthma. In the present paper, we retrospectively analyzed long-term asthma control by pranlukast treatment in patients treated with inhaled corticosteroids. We analyzed medical records and asthma diaries of 21 patients (9 males, 12 females) (52.1 +/- 3.5 years of age) with bronchial asthma who experienced increase of more than 10 L/min in peak expiratory flow in the first 4 weeks of treatment with pranlukast (450 mg/day) and were subsequently treated with pranlukast for more than 1 year. They all received inhaled corticosteroids (400-1600 microg/day of beclomethasone dipropionate or equivalent). We examined clinical control in terms of time course of self-monitored peak expiratory flow. During the analyzed period, the dose of inhaled corticosteroids was tapered in 4 patients, constant in 15 patients and increased in 2 patients. In 19 patients treated with unchanged or tapered dose of inhaled corticosteroids, improvement in the increase of mean PEF at 4-week treatment was maintained for 1 year. No difference in the add-on effect of pranlukast was observed in patients treated with less than 800 microg and more than or equal to 800 microg of inhaled corticosteroids. Four patients underwent reduction of inhaled corticosteroids in the analyzed period and PEF was well-maintained and even increased by pranlukast treatment. In 11 patients in whom data for 3 years were available, the improvement in PEF persisted for 3 years. Although the present investigation is a retrospective analysis, these data may suggest that pranlukast has no tachyphylaxis and its effect continues for more than 1 year.
Kazuo Yoneda, Kazuhito Rokutan, Yoichi Nakamura, Hiroaki Yanagawa, Shigetada Kondo and Saburo Sone : Stimulation of human bronchial epithelial cells by IgE-dependent histamine-releasing factor, American Journal of Physiology. Lung Cellular and Molecular Physiology, Vol.286, No.1, 174-181, 2004.
(要約)
An IgE-dependent histamine-releasing factor (HRF p23; also known as translationally controlled tumor protein or p23) stimulates the release of histamine, IL-4, and IL-13 from a subpopulation of highly allergic donor basophils. It has also been shown to act as a chemoattractant for eosinophils. To elucidate novel functions of HRF p23 in airway inflammation, we examined the effects of human recombinant HRF p23 (hrHRF) on bronchial epithelium and found that hrHRF stimulated the secretions of IL-8 and granulocyte/macrophage colony-stimulating factor by both primary cultures of human bronchial epithelial cells and BEAS-2B cells. In response to hrHRF, these cells induced IL-8 mRNA expression within 4 h. H2O2, but not IL-1 beta or tumor necrosis factor-alpha, stimulated secretion of HRF p23 by BEAS-2B cells, suggesting that oxidative stress may trigger the release of HRF p23 from bronchial epithelial cells. Bronchoalveolar lavage (BAL) from healthy volunteers contained only trivial or undetectable amounts of HRF p23. Significantly higher amounts of HRF p23 were recovered from BAL fluid taken from asthmatic patients, and the amounts of HRF p23 were further elevated in patients with idiopathic eosinophilic pneumonia. Our results demonstrate for the first time that HRF p23 can stimulate nonimmune epithelium. HRF p23 derived from bronchial epithelial cells may regulate complex cytokine networks in eosinophil-dependent inflammation of the human airway.
Toru Asano, Fumitaka Ogushi, Kenji Tani, Hiroyuki Tamiya, Yasuhiko Nishioka and Saburo Sone : Increased macrophage inflammatory protein-1α and -1β in BAL fluid of bronchiolitis obliterans organizing pneumonia, Respirology, Vol.8, No.4, 461-466, 2003.
(要約)
CC chemokines are mainly chemotactic for monocytes and lymphocytes. The aim of this study was to evaluate the involvement of the CC chemokines, macrophage inflammatory protein (MIP)-1alpha and MIP-1beta, in the pathogenesis of bronchiolitis obliterans organizing pneumonia (BOOP). The concentrations of MIP-1alpha and MIP-1beta in BAL fluid (BALF) obtained from patients with BOOP (n = 13) and control patients (CP, n= 18) were measured by enzyme-linked immunosorbent assay. MIP-1alpha in BALF was significantly higher in patients with BOOP (mean +/- SD; 123.8 +/- 98.0 pg/mL) than in CP (62.5 +/- 46.1 pg/mL). Significantly higher MIP-1beta was also detected in patients with BOOP (51.6 +/- 72.5 pg/mL) than in CP (6.4 +/- 3.7 pg/mL). The concentration of MIP-1alpha significantly correlated with the percentage of lymphocytes in BALF, and the concentration of MIP-1beta significantly correlated with the numbers of lymphocytes, neutrophils and eosinophils in BALF. Both MIP-1alpha and MIP-1beta in BALF were decreased after corticosteroid therapy and this was accompanied by decreased lymphocytes in BALF. This study suggests that MIP-1alpha and MIP-1beta may play important roles in the recruitment of immuno-inflammatory cells into the lungs, and may contribute to the pathogenesis of BOOP.
Seiji Yano, Helong Zhang, Masaki Hanibuchi, Toyokazu Miki, Hisatsugu Goto, Hisanori Uehara and Saburo Sone : Combined therapy with a new bisphosphonate, minodronate (YM529), and chemotherapy for multiple organ metastases of small cell lung cancer cells in severe combined immunodeficient mice., Clinical Cancer Research, Vol.9, No.14, 5380-5385, 2003.
(要約)
Lung cancer in the advanced stage frequently metastasizes to multiple organs, including the liver, lungs, lymph nodes, and bone. Bisphosphonates have been widely used to treat osteolytic bone metastasis in the past years; however, many studies have implicated that a single use of bisphosphonates could not prolong the survival of patients. In the present study, using a multiple-organ metastasis model of human lung cancer cells, we examined the effect of combined therapy with a new bisphosphonate (YM529) and etoposide (VP-16). Human small cell lung cancer (SBC-5) cells i.v. inoculated into natural killer cell-depleted severe combined immunodeficient mice metastasized to multiple organs, including the lungs, liver, kidneys, lymph nodes, and bone. SBC-5-bearing mice were treated with YM529 and/or VP-16 and sacrificed 5 weeks after tumor cell inoculation. Bone metastasis was assessed by X-ray photographs, and visceral metastasis was evaluated macroscopically. The number of osteoclasts in the bone lesions was examined by tartrate-resistant acid phosphatase staining. Monotherapy with YM529 suppressed the production of bone metastases, but not visceral metastasis. Histological analyses revealed that the number of osteoclasts in bone lesions was lower in YM526-treated mice, compared with control mice. VP-16 inhibited both bone metastasis and visceral (lung and liver) metastasis. However, neither YM529 alone nor VP-16 alone significantly prolonged the survival of SBC-5-bearing mice. Combined use of YM529 and VP-16 further inhibited the production of bone metastasis and significantly prolonged survival. Combined therapy with bisphosphonate and chemotherapy may be useful for small cell lung cancer patients with multiple organ metastases including bone metastasis.
T Sano, Y Nakamura, Hiroaki Yanagawa, H Bando, A Nii, S Yoshida and Saburo Sone : Add-on Effects of Suplatast Tosilate in Bronchial Asthma Patients Treated With Inhaled Corticosteroids, Lung, Vol.181, No.4, 227-235, 2003.
(要約)
Th2 cytokines play an important role in the pathogenesis of asthma. In the present study, we investigated the effect of suplatast tosilate, a selective Th2 cytokine inhibitor, on asthma control, in terms of subjective symptoms and pulmonary function in patients treated with inhaled corticosteroids. Thirty-eight patients with bronchial asthma being treated with inhaled corticosteroids were given suplatast tosilate (100 mg three times daily) for 12 weeks, in a multicenter setting. During the study period, other medications were continued. Morning and evening peak expiratory flow, asthma symptoms, blood eosinophil count and serum IgE levels were monitored. Suplatast tosilate treatment was associated with a significant improvement in mean morning peak expiratory flow (from 295 L/min to 348 L/min, P < 0.01) and evening peak expiratory flow (from 313 L/min to 357 L/min, P < 0.01). The mean daily variation in peak expiratory flow was significantly reduced (from 11.6% to 7.3%, P < 0.01) by suplatast tosilate treatment. The greatest improvement in peak expiratory flow was observed in patients whose blood eosinophil counts were decreased by suplatast tosilate treatment. Treatment with suplatast tosilate improved pulmonary function in patients with bronchial asthma. Our results suggest the therapeutic effects observed may occur through suppression of eosinophilic inflammation.
Hirohumi Dan, Kenji Tani, Kayoko Hase, Teruki Shimizu, Hiroyuki Tamiya, Yanjmaa Biraa, Luping Huang, Hiroaki Yanagawa and Saburo Sone : CD13/aminopeptidase N in collagen vascular diseases, Rheumatology International, Vol.23, No.6, 271-276, 2003.
(要約)
To determine the significance of CD13/aminopeptidase N in collagen vascular diseases (CVD), we examined its activity and expression in sera and disease sites of patients with CVD. Significantly higher aminopeptidase activity was detected in bronchoalveolar lavage fluid from patients with interstitial lung diseases due to rheumatoid arthritis (RA), polymyositis/dermatomyositis (PM/DM), systemic sclerosis (SSc), and Sjögren's syndrome than from control subjects. Increased aminopeptidase activity and increased expression of CD13/aminopeptidase N protein were found in alveolar macrophages from CVD patients with interstitial lung diseases. Significantly higher aminopeptidase activity was detected in pleural effusions from patients with systemic lupus erythematosus (SLE) than in transudate effusions. The mean aminopeptidase activity in synovial fluids from RA patients was significantly higher than from patients with osteoarthritis. The mean value of serum aminopeptidase activity was significantly higher in patients with SLE, RA, SSc, and PM/DM than in normal subjects. This study suggests that the activity of CD13/aminopeptidase N, locally produced in the disease site, is a useful marker for CVD and that CD13/aminopeptidase N may have an important role in the pathogenesis of CVD.
H Ogawa, N Nishimura, Yasuhiko Nishioka, M Azuma, Hiroaki Yanagawa and Saburo Sone : Adenoviral interleukin-12 gene transduction into human bronchial epithelial cells:up-regulation of pro-inflammatory cytokines and its prevention by corticosteroids, Clinical and Experimental Allergy, Vol.33, No.7, 921-929, 2003.
(要約)
One of the potential effects of IL-12 is to restore Th1/Th2 balance. Therefore, we investigated the possibility of developing a system for local delivery of IL-12 into the airways by examining protein expression in a human bronchial epithelial cell line (BEAS-2B) after adenoviral IL-12 gene transduction. The effects of dexamethasone on the gene-modified cells were also examined. Adenoviral vectors AxCAegfp and Ax1CIhp40ip35 were used to transduce enhanced green fluorescence protein and IL-12 genes, respectively, into BEAS-2B cells. Wild-type and IL-12 gene-transduced BEAS-2B cells were then incubated with or without dexamethasone, and concentrations of IL-12, IFN-gamma, IL-6, IL-8, granulocyte macrophage-colony stimulating factor and chemokines (TARC and RANTES) in the supernatant were measured by ELISA. IL-12 receptor expression was analysed by flow cytometry and RT-PCR. The efficiency of transgene expression in BEAS-2B cells at a multiplicity of infection of 30 was approximately 80%. Gene-modified BEAS-2B cells produced biologically active IL-12, regardless of dexamethasone treatment. While IL-12 gene transduction led to increased production of IL-6 and IL-8 by BEAS-2B cells, expressions of these proteins were suppressed by dexamethasone. Addition of exogenous IL-12 failed to augment BEAS-2B cell IL-6 and IL-8 production, and IL-12 receptor expression by BEAS-2B cells was not detected. Our findings suggest that adenoviral IL-12 gene transduction may be effective in inducing IL-12 expression in the airways, and could be a potential approach in the management of bronchial asthma.
Seiji Yano, Yasuhiko Nishioka, Hisatsugu Goto and Saburo Sone : Molecular mechanisms of angiogenesis in non-small cell lung cancer, and therapeutics targeting related molecules., Cancer Science, Vol.94, No.6, 479-485, 2003.
(要約)
Angiogenesis, neovascularization from pre-existing vasculature, is necessary to supply oxygen and nutrition for tumor growth in both primary and distant organs. It consists of sprouting and non-sprouting (the enlargement, splitting, and fusion of pre-existing vessels) processes, and both can occur concurrently. Growth of solid tumors, including non-small cell lung cancer (NSCLC), is usually dependent on angiogenesis, which is regulated by complex mechanisms involving various angiogenesis-related molecules. Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF), one of the most potent angiogenic molecules, regulates both angiogenesis and vascular permeability, and hence promotes tumor progression and development of malignant pleural effusions in NSCLC. Signals via epidermal growth factor receptor (EGFR) promote not only the tumor cell cycle, but also the process of angiogenesis. Therefore, these molecules are potential targets for anti-tumor vasculature therapy. Many agents targeting tumor vasculature have been developed, and several compounds have shown anti-tumor potential in preclinical studies. Their efficacy against NSCLC is currently being evaluated in clinical trials.
Hiroya Tada, Fumitaka Ogushi, Kenji Tani, Yasuhiko Nishioka, Jun-ya Miyata, Keiko Sato, Toru Asano and Saburo Sone : Increased binding and chemotactic capacities of PDGF-BB on fibroblasts in radiation pneumonitis, Radiation Research, Vol.159, No.6, 805-811, 2003.
(要約)
Although pulmonary fibrosis is a frequent and serious consequence of radiotherapy for thoracic malignant diseases such as lung cancer, the pathogenesis of this radiation-induced lung disorder remains unclear. To clarify the mechanisms underlying radiation pneumonitis and pulmonary fibrosis, we investigated the expression of platelet-derived growth factor receptor (PDGFR) on fibroblasts obtained from irradiated rat lungs and on control fibroblasts. Whole lungs of male Wistar rats were irradiated with a single dose of 15 Gy, and lung fibroblasts were isolated at 4 weeks after the irradiation. The chemotactic response of irradiated lung fibroblasts to PDGF-BB was significantly higher than that of control lung fibroblasts, whereas there was no significant difference between irradiated lung fibroblasts and control lung fibroblasts in the response to PDGF-AA. Receptor binding assay showed more specific binding sites for PDGF-BB on irradiated lung fibroblasts than on control lung fibroblasts, and the displacement of (125)I-labeled PDGF binding to fibroblasts by unlabeled PDGF showed that (125)I-labeled PDGF-BB was displaced by PDGF-BB but not by PDGF-AA. These results suggest that the increased binding sites for PDGF-BB on irradiated lung fibroblasts correspond mainly to PDGFRB. Scatchard analysis of the saturation data demonstrated an approximately twofold increase both in the number of PDGF-BB binding sites and in the binding affinity in irradiated lung fibroblasts compared to that in control lung fibroblasts. Those results suggest that the increased chemotactic response of irradiated lung fibroblasts to PDGF-BB is related to the overexpression of PDGFRB, which may have an important role in the pathogenesis of radiation-induced pneumonitis and pulmonary fibrosis.
Souji Kakiuchi, Yataro Daigo, Tatsuhiko Tsunoda, Seiji Yano, Saburo Sone and Yusuke Nakamura : Genome-Wide Analysis of Organ-Preferential Metastasis of Human Small Cell Lung Cancer in Mice, Molecular Cancer Research : MCR, Vol.1, No.7, 485-499, 2003.
(要約)
Although a number of molecules have been implicated in the process of cancer metastasis, the organ-selective nature of cancer cells is still poorly understood. To investigate this issue, we established a metastasis model in mice with multiple organ dissemination by i.v. injection of human small cell lung cancer (SBC-5) cells. We analyzed gene-expression profiles of 25 metastatic lesions from four organs (lung, liver, kidney, and bone) using a cDNA microarray representing 23,040 genes and extracted 435 genes that seemed to reflect the organ specificity of the metastatic cells and the cross-talk between cancer cells and microenvironment. Furthermore, we discovered 105 genes that might be involved in the incipient stage of secondary-tumor formation by comparing the gene-expression profiles of metastatic lesions classified according to size (<1 or >2 mm) as either "micrometastases" or "macrometastases." This genome-wide analysis should contribute to a greater understanding of molecular aspects of the metastatic process in different microenvironments, and provide indicators for new strategies to predict and prevent metastasis.
Yasuhiko Nishioka, Hua Wen, Kayo Mitani, Paul D. Robbins, Michael T. Lotze, Saburo Sone and Hideaki Tahara : Differential effects of IL-12 on the generation of alloreactive CTL mediated by murine and human dendritic cells: a critical role for nitric oxide, Journal of Leukocyte Biology, Vol.73, No.5, 621-629, 2003.
(要約)
We examined the mechanisms involved in interleukin (IL)-12-mediated suppression of cellular immunity in mice using allogeneic mixed leukocyte reaction (MLR) stimulated by dendritic cells (DCs) in vitro and compared the effect of IL-12 on MLR in mice and humans. Although IL-12 stimulated human MLR, the addition of IL-12 or interferon-gamma (IFN-gamma) resulted in a dose-dependent suppression of MLR in mice. The treatment with N(G)-monomethyl-L-arginine (L-NMMA) completely abrogated IL-12- and IFN-gamma-mediated suppression of MLR in mice. Furthermore, IL-12 enhanced the alloreactive cytolytic T lymphocyte (CTL) induction in human MLR, whereas the addition of L-NMMA was required to generate alloreactive CTLs in the presence of IL-12 in mice. Nitric oxide (NO) was detected only in mouse MLR. Murine DCs could produce NO, but neither human CD34(+) cell- nor monocyte-derived DCs produced a detectable amount of NO. These results suggest that NO produced by DCs might play an important role in IL-12-mediated immune suppression in mice but not in humans.
Seiji Yano, Kanematsu Takahori, Toyokazu Miki, Aono Yoshinori, Masahiko Azuma, Akihiko Yamamoto, Hisanori Uehara and Saburo Sone : A report of two bronchioloalveolar carcinoma cases which were rapidly improved by treatment with the epidermal growth factor receptor tyrosine kinase inhibitor ZD1839 ("Iressa")., Cancer Science, Vol.94, No.5, 453-458, 2003.
(要約)
Bronchioloalveolar carcinoma (BAC), a form of pulmonary adenocarcinoma, presents unique clinical features, such as endobronchial spread and bronchorrhea in advanced stages. The prognosis for BAC patients in advanced stages is poor, as is the case for patients with other non-small-cell lung cancer (NSCLC) types, because of low susceptibility to conventional chemotherapy. Recently, an orally active, selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (EGFR-TKI), ZD1839 ("Iressa"), has been investigated in phase II clinical studies (IDEAL 1 and IDEAL 2) as monotherapy against chemotherapy-refractory NSCLC, and provided clinically significant antitumor activity. In this study, we examined the therapeutic efficiency of ZD1839 in chemotherapy-refractory BAC patients with bronchorrhea. Two female BAC patients with bronchorrhea were treated once daily with ZD1839 (250 mg/day). In both cases, serous sputum production was dramatically reduced within 3 days of starting the treatment, and hypoxia and radiographic signs of bilateral lung consolidation were visibly improved within 7 days. Following more than 8 months of treatment, no evidence of recurrence or severe adverse events has been observed. These results suggest that this selective EGFR-TKI, ZD1839, may be a powerful agent for treatment of chemotherapy-refractory BAC patients with bronchorrhea.
Akihiko Yamamoto, Seiji Yano, Minoru Shiraga, Hirohisa Ogawa, Hisatsugu Goto, Toyokazu Miki, Helong Zhang and Saburo Sone : A third-generation matrix metalloproteinase (MMP) inhibitor (ONO-4817) combined with docetaxel suppresses progression of lung micrometastasis of MMP-expressing tumor cells in nude mice., International Journal of Cancer, Vol.103, No.6, 822-828, 2003.
(要約)
The lung is the common target organ of hematogenous metastasis that restricts the prognosis of cancer patients. MMPs play a pivotal role in metastasis by promoting tumor invasion and angiogenesis; therefore, a large number of MMPIs have been developed. Our purpose was to determine the therapeutic efficacy of a selective-spectrum MMPI, ONO-4817 (inhibits MMP-2 and MMP-9 but not MMP-1), against established lung micrometastasis in combination with a cytotoxic anticancer drug, DOC, in a nude mouse model. Human non-small cell lung cancer PC14PE6 (adenocarcinoma) or H226 (squamous cell carcinoma) cells, expressing MMP-2, MMP-9 and/or MMP-1, were injected i.v. into nude mice on day 0. Mice received a single injection of DOC on day 7 (after establishment of micrometastasis) and/or ONO-4817 mixed with food from day 7 to the end of experiments. Monotherapy with ONO-4817 or DOC inhibited formation of lung metastasis by PC14PE6 and H226 cells. In addition, combined use of ONO-4817 with DOC significantly suppressed the tumor burden of H226 and PC14PE6 cells in the lung and prolonged the survival of PC14PE6-bearing mice compared to ONO-4817 or DOC alone. These therapies did not affect the body weight or food intake of tumor-bearing mice. FIZ revealed that lung lesions, but not nontumor parenchyma of the lung, expressed gelatinolytic activity and that treatment with ONO-4817 abrogated the gelatinolytic activity in lung lesions. These results suggest that the combined use of MMPIs with cytotoxic anticancer drugs may be helpful in the control of established lung micrometastasis by tumor cells expressing MMPs.
Kayo Mitani, Yasuhiko Nishioka, Kazue Yamabe, Hirohisa Ogawa, Toyokazu Miki, Hiroaki Yanagawa and Saburo Sone : Soluble Fas in malignant pleural effusion and its expression in lung cancer cells, Cancer Science, Vol.94, No.3, 302-307, 2003.
(要約)
Soluble Fas (sFas) has the ability to block Fas-mediated apoptosis, suggesting that sFas at tumor sites might inhibit tumor cell-killing by immune effector cells. We examined the sFas level in pleural effusion associated with lung cancer. The level of sFas in malignant pleural effusion was significantly higher than those in transudate and tuberculous pleural effusion. There was no significant difference in the sFas concentration among various histological types of lung cancer. The cytotoxicity mediated by anti-Fas agonistic antibody against Jurkat cells was inhibited by the addition of malignant pleural effusion, being inversely correlated with the sFas concentration. When Fas expression was examined using flow cytometry, eight of ten (80%) lung cancer cell lines expressed cell surface Fas. On the other hand, sFas protein and mRNA were detected in six of ten (60%) lung cancer cell lines, but there was no correlation between Fas and sFas expression. Furthermore, although the expressions of Fas and sFas were clearly detected in tumor cells derived from malignant effusion, the sFas expression was down-regulated in an in vitro culture. These results suggest that sFas in malignant pleural effusion is at least in part produced by lung cancer cells, and might play a role in local immunosuppression by tumor cells.
Seiji Yano, Hiroshi Nokihara, Akihiko Yamamoto, Hisatsugu Goto, Hirohisa Ogawa, Takanori Kanematsu, Toyokazu Miki, Hisanori Uehara, Yasuo Saijo, Toshihiro Nukiwa and Saburo Sone : Multifunctional interleukin-1beta promotes metastasis of human lung cancer cells in SCID mice via enhanced expression of adhesion-, invasion- and angiogenesis-related molecules., Cancer Science, Vol.94, No.3, 244-252, 2003.
(要約)
We examined whether interleukin-1 (IL-1), a multifunctional proinflammatory cytokine, progresses or regresses metastasis of lung cancer. Exogenous IL-1beta enhanced expression of various cytokines (IL-6, IL-8, and vascular endothelial growth factor (VEGF)) and intracellular adhesion molecule-1 (ICAM-1) by A549, PC14, RERF-LC-AI, and SBC-3 cells expressing IL-1 receptors. A549 cells transduced with human IL-1beta-gene with the growth-hormone signaling-peptide sequence (A549/IL-1beta) secreted a large amount of IL-1beta protein. Overexpression of IL-1beta resulted in augmentation of expression of the cytokines, ICAM-1, and matrix metalloproteinase-2 (MMP-2). A549/IL-1beta cells intravenously inoculated into severe combined immunodeficiency (SCID) mice distributed to the lung more efficiently and developed lung metastasis much more rapidly than did control A549 cells. Treatment of SCID mice with anti-IL-1beta antibody inhibited formation of lung metastasis by A549/IL-1beta cells. Moreover, A549/IL-1beta cells inoculated in the subcutis grew more rapidly, without necrosis, than did control A549 cells, which produced smaller tumors with central necrosis, suggesting involvement of angiogenesis in addition to enhanced binding in the high metastatic potential of A549/IL-1beta cells. Histological analyses showed that more host-cell infiltration, fewer apoptotic cells, more vascularization, and higher MMP activity were observed in tumors derived from A549/IL-1beta cells, compared with tumors derived from control A549 cells. These findings suggest that IL-1beta facilitates metastasis of lung cancer via promoting multiple events, including adhesion, invasion and angiogenesis.
Mari Miki, Yoichi Nakamura, Akira Takahashi, Yutaka Nakaya, Hiroshi Eguchi, Tsukio Masegi, Kazuo Yoneda, Susumu Yasuoka and Saburo Sone : Effect of human airway trypsin-like protease on intracellular free Ca2+ concentration in human bronchial epithelial cells, The Journal of Medical Investigation : JMI, Vol.50, No.1-2, 95-107, 2003.
(要約)
It has been shown that human airway trypsin-like protease (HAT) is localized in human bronchial epithelial cells (HBEC), and trypsin activates protease-activated receptor-2 (PAR-2). Activation of PAR-2 activates G-protein followed by an increase of intracellular free Ca2+, [Ca2+]in. This study was undertaken to clarify whether HAT can activate PAR-2 in HBEC or not. RT-PCR showed that HAT mRNA is expressed in HBEC, and PAR-2 mRNA is the most strongly expressed of the known PARs in HBEC. Both PAR-2 agonist peptide (PAR-2 AP) and HAT increased [Ca2+]in in HBEC in a biphasic fashion; a prompt, sharp increase (peak I) and a sustained low plateau (peak II). PAR-2 AP over 100-200 microM and HAT over 200-300 mU/ml (0.08-0.12 microM) induced both peak I and II, and PAR-2 AP below 100 microM and HAT below 200 mU/ml induced only peak II. Both PAR-2 AP-induced and HAT-induced peak I were induced by Ca2+ mobilization from intracellular stores, because they appeared even in Ca2+-free medium. Both PAR-2 AP-induced and HAT-induced peak II were induced by an influx of extracellular Ca2+, because they were abolished in Ca2+-free medium. The Ca2+ response to HAT was desensitized by exposure of HBEC to PAR-2 AP. These results indicate that HBEC have a functional PAR-2, and HAT regulates cellular functions of HBEC via activation of PAR-2.
Hiroaki Yanagawa, Minoru Shiraga, Youichi Nakamura, Masahiko Azuma, Kazuo Yoneda, Hirohisa Ogawa, Chikato Kitamuro, Kenichi Maeda, Mari Miki, Yuka Matsumori, Akemi Sugita, Sakiko Hashimoto, Chie Hara, Kenji Tani and Saburo Sone : Inhaled steroid therapy and hospitalization for bronchial asthma:trend in Tokushima University Hospital, The Journal of Medical Investigation : JMI, Vol.50, No.1,2, 72-77, 2003.
(要約)
With the recognition that airway inflammation is present even in patients with mild bronchial asthma, therapy with inhaled corticosteroids is now indicated in various stages of patients. In the present article, we retrospectively examined the prescriptions for inhaled corticosteroids and other drugs for the treatment of outpatients with bronchial asthma at Tokushima University Hospital. We also analyzed asthma control in these patients, in terms of the incidence of emergency consultations and hospitalizations due to asthma exacerbations. To analyze the recent trend, the patients observed from 1998 to 2000 (recent years) were included, and for control purpose, those in 1990 and 1991 (earlier years) were also included. The percentage of patients treated with inhaled corticosteroids remarkably increased in recent years (mean; 81.3%) compared to earlier years (mean; 23.5%). In contrast, the usage of oral corticosteroids, oral xanthine derivatives, beta-adrenergic receptor agonists and anti-allergic agents tended to decrease in the 10 years period. After the introduction in 1995, considerable patients up to 25% have been treated with anti-leukotrienes. Emergency consultations decreased in recent years (mean; 0.18/patient/year) compared to earlier years (mean; 0.79/patient/year). Emergency hospitalizations also decreased in recent years (mean; 0.043/patient/year) compared to earlier years (mean; 0.23/patient/ year). In the present study, spread of inhaled corticosteroid therapy and decline in incidence of emergency consultation and hospitalization were simultaneously observed at Tokushima University Hospital, and the former has, at least in part, a contribution to the latter.
Takanori Kanematsu, Seiji Yano, Hisanori Uehara, Yoshimi Bando and Saburo Sone : Phosphorylation, but not overexpression, of epidermal growth factor receptor is associated with poor prognosis of non-small cell lung cancer patients, Oncology Research, Vol.13, No.5, 289-298, 2003.
(要約)
Epidermal growth factor receptor (EGFR) is commonly overexpressed in non-small cell lung cancer (NSCLC) and its tyrosine kinase phosphorylation is thought to be an ideal target in the treatment of patients with NSCLC. In the present study, we examined surgically obtained specimens from a series of 36 NSCLC patients for expression of EGFR, phosphorylated EGFR (p-EGFR), and HER2 by immunohistochemistry, and also examined the correlation with clinical characteristics. The positive rate of EGFR, p-EGFR, and HER2 was 97.2%, 44.4%, and 88.6%, respectively, and the overexpression rate was 80.6%, 0.0%, and 27.8%, respectively. EGFR overexpression and phosphorylation were seen at almost the same rate in each histological type of squamous and nonsquamous cell carcinoma (squamous vs. nonsquamous; 78.6% vs. 81.8% for EGFR, 35.7% vs. 50.0% for p-EGFR), while HER2 overexpression was seen less frequently in squamous cell carcinoma than in nonsquamous cell carcinoma (0.0% vs. 45.5%, P = 0.003). Univariate analysis revealed that EGFR overexpression was related to good performance status (P = 0.038) but not related to EGFR phosphorylation. EGFR phosphorylation was correlated to short time to progression (TTP) (P = 0.002) and poor prognosis (P = 0.002), although EGFR overexpression, HER2 overexpression, or EGFR-HER2 coexpression were not correlated to TTP or survival. Bivariate analysis showed EGFR phosphorylation was related to short TTP and poor prognosis both in early and advanced stages. Multivariate analyses confirmed that clinical stage, performance status, and p-EGFR expression were independently associated with increasing risk of short TTP and poor prognosis. These results suggest that phosphorylation, but not overexpression, of EGFR may be an important predictor for clinical outcome of NSCLCs.
Helong Zhang, Seiji Yano, Toyokazu Miki, Hisatsugu Goto, Takanori Kanematsu, Hiroaki Muguruma, Hisanori Uehara and Saburo Sone : A novel bisphosphonate minodronate (YM529) specifically inhibits osteolytic bone metastasis produced by human small-cell lung cancer cells in NK-cell depleted SCID mice., Clinical & Experimental Metastasis, Vol.20, No.2, 153-159, 2003.
(要約)
In the present study, we examined the effects of a newly developed bisphosphonate, minodronate (YM529), on osteolytic bone metastasis caused by lung cancer. Human small-cell lung cancer (SBC-5) cells, injected intravenously into natural killer cell-depleted SCID mice, produced radiologically detectable bone metastasis by day 18 and macroscopically visible visceral metastases (lung, liver, kidney, systemic lymph node) by day 35. Prophylactic treatment with YM529 on day 1 significantly inhibited the formation of osteolytic bone metastasis evaluated on X-ray photographs in a dose-dependent manner. In addition, treatment with YM529 after establishment of bone metastasis (on day 21) also inhibited bone metastasis, although the treatment was more effective when started earlier. Single administration was as effective as repeated treatment, suggesting a sustained inhibitory effect of YM529 on bone metastasis. YM529 reduced the number of osteoclasts in the bone metastatic lesions in vivo, but had no effect on the proliferation or cytokine production of SBC-5 cells in vitro. These results suggest that YM529 is a potent inhibitor of bone metastasis of human lung cancer, probably by suppressing osteoclastic bone resorption. In contrast, treatment with YM529 had no effect on visceral metastasis, even if started on day 1, and did not prolong the survival of the mice. Therefore, development of a combined modality is necessary for prolonging the survival of small-cell lung cancer patients with multiple-organ metastasis.
Minoru Shiraga, Seiji Yano, Akihiko Yamamoto, Hirohisa Ogawa, Hisatsugu Goto, Toyokazu Miki, Keisuke Miki, Helong Zhang and Saburo Sone : Organ Heterogeneity of Host-derived Matrix Metalloproteinase Expression and Its Involvement in Multiple-Organ Metastasis by Lung Cancer Cell Lines, Cancer Research, Vol.62, No.20, 5967-5973, 2002.
(要約)
Cancer metastasis is tightly regulated by the interaction of tumor cells and host organ microenvironments. Matrix metalloproteinases (MMPs), produced by both tumor cells and host stromal cells, play a central role in tumor invasion and angiogenesis. We determined whether metastatic potential of lung cancer to multiple organs is dependent solely on the expression of MMPs by tumor cells, using two metastasis models of human lung cancer cell lines expressing various levels of MMPs and a MMP inhibitor (ONO-4817). In the lung metastasis model, tumor cells (PC14, PC14PE6, H226, A549) inoculated i.v. into nude or SCID mice metastasized only in the lung. In the multiple-organ metastasis model, tumor cells (RERF-LC-AI, SBC-3/DOX, H69/VP, which express low levels of MMPs) inoculated i.v. into natural killer cell-depleted SCID mice metastasized into the liver, kidneys, and systemic lymph nodes. Film in situ zymography analysis revealed that the nontumor parenchyma of the lung had no gelatinolytic activity, whereas gelatinolytic activity of the liver and kidney was high and low, respectively. In the lung metastasis model, gelatinolytic activity of lung nodules directly correlated with the in vitro expression of MMP-2 and MMP-9 by tumor cells. Inhibition of MMP activity by ONO-4817 suppressed lung metastasis by the cell lines that expressed MMPs, but not those that did not express MMP, via the inhibition of MMP activity of lung tumors. In the multiple-organ metastasis model, liver parenchyma, but not liver nodules, showed gelatinolytic activity. The MMP inhibition reduced metastasis to the liver, but not to the kidney or lymph nodes, via inhibition of MMP activity of liver parenchyma. These findings suggest that MMP expression varies among the host organ microenvironments and that stromal MMPs may promote metastasis of lung cancer. Therefore, antimetastatic effects based on MMP inhibition may be dependent on MMPs derived not only from tumor cells but also from organ-specific microenvironments.
Teruki Shimizu, Kenji Tani, Kayoko Hase and Saburo Sone : CD13/aminopeptidase N Induced Lymphocyte Involvement in Inflamed Joints of Rheumatoid Arthritis, Arthritis and Rheumatism, Vol.46, No.9, 2330-2338, 2002.
(要約)
We previously showed that CD13/aminopeptidase N (EC 3.4.11.2) induces chemotactic migration of T lymphocytes by its enzymatic activity. In this study, we examined the role of CD13/aminopeptidase N in lymphocyte involvement in rheumatoid arthritis (RA). Synovial fluids were obtained from 27 RA patients and 6 osteoarthritis (OA) patients. Synovial tissue specimens were obtained from 3 RA patients and 3 OA patients. Protease activity of aminopeptidase in synovial fluids and synovial fibroblasts was assayed fluorometrically using the specific substrate. Expression of CD13/aminopeptidase N in synovial fibroblasts was determined by flow cytometry analyses, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR). The mean value of aminopeptidase activity in synovial fluid samples from RA patients was significantly higher than that in samples from OA patients. Increased enzymatic activity of aminopeptidase was detected on synovial fibroblasts from RA patients compared with those from OA patients. Flow cytometry showed that the expression of CD13/aminopeptidase N on synovial fibroblasts from RA patients was higher than the expression on synovial fibroblasts from OA patients, and Western blots and RT-PCR showed that synovial fibroblasts from RA patients contained a greater amount of CD13/aminopeptidase N. The activity of CD13/aminopeptidase N correlated significantly with lymphocyte counts in synovial fluids from RA patients. Synovial fluids from RA patients in which high aminopeptidase activity was detected contained considerable chemotactic activity for lymphocytes, and bestatin, a specific inhibitor of aminopeptidases, partially inhibited the chemotactic activity. CD13/aminopeptidase N may participate in the mechanism of lymphocyte involvement in inflamed joints of RA patients as a lymphocyte chemoattractant.
Hisatsugu Goto, Seiji Yano, Helong Zhang, Yuka Matsumori, Hirohisa Ogawa, David C. Blakey and Saburo Sone : Activity of a New Vascular Targeting Agent, ZD6126, in Pulmonary Metastases by Human Lung Adenocarcinoma in Nude Mice, Cancer Research, Vol.62, No.13, 3711-3715, 2002.
(要約)
ZD6126 (ANG453) is a novel vascular targeting agent that selectively disrupts the cytoskeleton of endothelial cells in tumor. In mouse s.c. xenograft models, ZD6126 was found to induce selective occlusion of tumor blood vessels, cessation of tumor blood flow, and death of tumor cells because of the starvation of oxygen and nutrition. Here, we investigated whether ZD6126 inhibited the metastatic formation of human non-small cell lung cancer cells. PC14PE6 (adenocarcinoma) and H226 (squamous cell carcinoma) cells were injected into the tail vein of nude mice, and lung metastases were estimated. ZD6126 treatment involved either a single dose on 24 h before killing or daily doses from day 14 until the end of the experiment. Single treatment with i.p. injection of 200 mg/kg ZD6126 caused bleeding and necrotic changes in the tumor by 24 h. Histological analysis revealed that apoptotic tumor cells were markedly increased in the ZD6126-treated group. Moreover, ZD6126 induced the apoptosis of CD31-positive vascular endothelial cells in tumors but not in the normal lung parenchyma. When mice were treated daily with 100 mg/kg ZD6126 from day 14 until the end of the experiment, the lung weight was significantly less in the ZD6126-treated group than that of the control group, despite no difference in the number of metastatic nodules. These data suggest that ZD6126 could demonstrate its antitumor activity against both already established and early phase of lung cancer metastasis by causing the selective apoptosis of tumor endothelial cells and destruction of the tumor vasculature.
Kenji Tani, Teruki Shimizu, Yumi Motoki and Saburo Sone : Chemokines in synovial inflammation in rheumatoid arthritis: basic and clinical aspects, Modern Rheumatology, Vol.12, No.2, 93-99, 2002.
(要約)
Abstract Rheumatoid arthritis (RA) is a chronic, multisystem autoimmune disease characterized by persistent synovitis. Since chemotactic cytokines (chemokines) may play critical roles in the recruitment of leukocytes in RA, analyses of chemokines and their receptors should provide insight into events in synovial inflammation in RA. The production of chemokines is regulated by cytokines such as tumor necrosis factor (TNF)-α produced in the inflamed joint, suggesting that the efficacy of anti-TNF-α therapy is mediated at least partly by the reduction of chemokine production. Chemokines have a role in joint inflammation not only by inducing leukocyte chemotaxis, but also by activating immune cells and angiogenesis. The pathogenesis of RA has been shown to be mediated by Th1-type T cells, because Th1-related chemokine receptors are preferentially expressed on cells in synovial fluid and synovial tissue. Accordingly, antichemokine therapy may be important as a possible new approach to therapeutic intervention in RA.
Hiroyoshi Sei, Atsuko Sano, Hiromi Ohno, Kazue Yamabe, Yasuhiko Nishioka, Saburo Sone and Yusuke Morita : Age-related changes in control of blood pressure and heart rate during sleep in the rat, Sleep, Vol.25, No.3, 279-285, 2002.
(要約)
The aim of this study was to determine age-related changes in the control of mean arterial pressure (MAP) and heart rate (HR) during sleep, and its relationship to the baroreflex in aging. MAP, HR, body temperature (TP), spontaneous activity (ACT), and sleeping/waking duration were monitored for 24 hours in groups of young (10-12 wk old) and old (23-24 mo old) rats. The sleep laboratory at the University of Tokushima. Subjects were 8 young (10-12 wk old) and 7 old (23-24 mo old) Wistar rats. Reflex control of HR was evaluated by examining various pressure responses to an intravenous bolus injection of phenylephrine and sodium nitroprusside. MAP and TP were recorded by a radiotelemetry system. HR was detected from the AP signal. ACT was counted by a photo-sensor system. In the case of old rats, the sensitivity of baroreflex control of HR was significantly depressed, and the spontaneous increase of MAP and HR during REM sleep and the MAP drop at the end of REM sleep were significantly enhanced. The old rats showed no large deterioration of the circadian profiles of MAP, HR, TP, and the amount of sleep. The baroreflex dysfunction is considered to appear in an early stage of the aging process, and to affect the control of MAP and HR during sleep.
Takashi Itokawa, Hiroki Nokihara, Yasuhiko Nishioka, Saburo Sone, Yukihide Iwamoto, Yuji Yamada, Julie Cherrington, Gerald McMahon, Masabumi Shibuya, Michihiko Kuwano and Mayumi Ono : Antiangiogenic Effect by SU5416 Is Partly Attributable to Inhibition of Flt-1 Receptor Signaling, Molecular Cancer Therapeutics, Vol.1, No.5, 295-302, 2002.
(要約)
Interaction between vascular endothelial growth factor (VEGF) and its cognate receptors, KDR/Flk-1 and Flt-1, of vascular endothelial cells is expected to induce an angiogenesis "switch" in tumors and other angiogenesis-associated diseases. SU5416, a selective inhibitor of the KDR/Flk-1 tyrosine kinase, is known to be a potent inhibitor of tumor angiogenesis. In this study, we first observed that SU5416 inhibited Flt-1 tyrosine kinase activity at similar doses, in addition to inhibiting KDR/Flk-1 tyrosine kinase activity in response to VEGF. SU5416 inhibited cell migration of human vascular endothelial cells expressing both Flt-1 and KDR in response to VEGF and also inhibited the cell migration in response to placenta growth factor (PIGF), a specific ligand for Flt-1. Chemotaxis of monocytes expressing only Flt-1 was also inhibited by SU5416 in a dose-dependent manner. Moreover, SU5416 was found to inhibit tyrosine kinase of Flt-1 in response to PIGF in vitro. And angiogenesis induced by PIGF in a Matrigel plug assay was inhibited by administration of SU5416. The antiangiogenic effects by this VEGF receptor-targeting compound appeared to be mediated through interference not only with KDR/Flk-1 but also with Flt-1. Cell migration of vascular endothelial cells and monocytic cells through Flt-1, thus, might play a key role in VEGF-induced tumor angiogenesis in concert with KDR/Flk-1.
Eiji Takeuchi, Hiroaki Yanagawa, Yoshihiro Suzuki, Kunihiro Shinkawa, Yasukazu Ohmoto, Hiroyasu Bando and Saburo Sone : IL-12-induced production of IL-10 and interferon-γ by mononuclear cells in lung cancer-associated malignant pleural effusions, Lung Cancer, Vol.35, No.2, 171-177, 2002.
(要約)
Interleukin (IL)-12 enhances natural killer (NK) activity and induces interferon gamma (IFN-gamma) production. Recently, it was shown that IL-12 induces IL-10 production by human T cells and NK cells, as a negative feedback for IL-12-induced immune responses. In the present study, in order to investigate the functions of host immune cells existing in contact with cancer cells, we examined the effect of IL-12 on the induction of non-major histocompatibility complex (MHC)-restricted killer activity and of IFN-gamma and IL-10 production by pleural and peripheral blood mononuclear cells (MNC), isolated from 40 lung cancer patients and 20 control subjects. IL-12 induced significant killer activity in pleural MNC from lung cancer patients, as well as those in peripheral blood, against a small cell lung cancer cell line (SBC-3). In lung cancer patients, pleural MNC incubated with IL-12 produced more IFN-gamma than blood MNC. In addition, when stimulated with both IL-12 and IL-2, pleural MNC produced more IL-10 than blood MNC. This is the first study reporting that MNC from pleural effusions of patients with lung cancer can produce both type 1 (IFN-gamma) and type 2 (IL-10) cytokines following exposure to IL-2 and IL-12. These observations suggest that control of IL-10 production at the microenvironment level may be important for the efficacy of human lung cancer immunotherapy with IL-12.
(キーワード)
Adult / Aged / Aged, 80 and over / Angiogenesis Inhibitors / Carcinoma, Small Cell / Cell Communication / Female / Humans / Interferon-gamma / Interleukin-10 / Interleukin-12 / Leukocytes, Mononuclear / Lung Neoplasms / Male / Middle Aged / Pleural Effusion / Tumor Cells, Cultured
Hirohisa Ogawa, Naoki Nishimura, Yasuhiko Nishioka, Masahiko Azuma, Hiroaki Yanagawa and Saburo Sone : Interleukin(IL)-12 gene transduction and its functional expression into human bronchial epithelial cells (BEAS-2B) by adenovirus vector, The Journal of Medical Investigation : JMI, Vol.49, No.1,2, 74-82, 2002.
(要約)
Interleukin (IL)-12 is known as a cytokine that augments the Th1 type response. Especially in allergic diseases such as a bronchial asthma, IL-12 induced restoration of the balance of the Th1/Th2 type immune response is an attractive strategy. In this study, the functional properties of the human bronchial epithelial cell line (BEAS-2B) transduced by an adenoviral vector encoding the human IL-12 gene were examined. Adenovirus vectors, AxCAegfp and Ax1CIhp40ip35 were transduced into BEAS-2B cells. Wild and gene-transduced BEAS-2B cells were incubated and the concentrations of IL-12 and IFN-gamma produced by co-cultured lymphocytes in the supernatant were measured using ELISA. The expressions of surface adhesion molecules, such as CD54 and CD106 were analyzed using flow cytometry. The efficiency of transgene expression of BEAS-2B cells was in a multiplicity of infection (MOI)-dependent manner and at an MOI of 30, the efficiency was approximately 80%. The gene-modified BEAS-2B cells produced biologically active IL-12 in dose- and time-dependent manners. IL-12 gene transduction did not significantly affect the expression of adhesion molecules (CD 54, CD106 and HLA-A,B,C) by BEAS-2B cells. These results suggest that the IL-12 gene may be successfully transduced into human bronchial epithelial cells by adenoviral vector to express IL-12 activity in vivo.
Yasuhiko Nishioka, Wen Hua, Naoki Nishimura and Saburo Sone : Genetic modification of dendritic cells and its application for cancer immunotherapy, The Journal of Medical Investigation : JMI, Vol.49, No.1,2, 7-17, 2002.
(要約)
Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs). DCs pulsed with peptides of tumor-associated antigens (TAA) and tumor lysate have been used in cancer immunotherapy. An early clinical study demonstrated the safety of the use of DCs, but the clinical response was not sufficient. The gene-modification of DCs with TAA and soluble factor genes such as cytokine and chemokine genes has been examined to enhance the antigen-presenting capacity of DCs. Viral vectors including retroviruses and adenoviruses have been reported to be useful to obtain a sufficient transduction efficiency into DCs. TAA gene-transduced DCs could have several advantages compared with TAA peptide-pulsed DCs as follows: 1) The use of TAA gene-modified DCs are not restricted by MHC haplotypes. 2) The gene transduction with TAA genes is likely to present the unknown TAA peptides on DCs. 3) The gene-modified DCs show the prolonged presentation of TAA peptides. The transduction of DCs with cytokine genes including IL-12 and GM-CSF have also been reported to argument the antitumor effects of DCs. Although the results in the experimental systems were promising, the clinical application of gene-modified DCs includes several problems such as the standardization of methods of manipulation and gene-transduction of DCs. Approaches to solve them require further studies.
Luping Huang, Kenji Tani, Fumitaka Ogushi, Hirohisa Ogawa, Teruki Shimizu, Yumi Motoki, Hiroki Moriguchi and Saburo Sone : Role of CD13/Aminopeptidase N in Rat Lymphocytic Alveolitis Caused by Thoracic Irradiation, Radiation Research, Vol.157, No.2, 191-198, 2002.
(要約)
CD13/aminopeptidase N is a cell surface glycoprotein that is widely distributed in a variety of mammalian cells. It was recently shown to have chemotactic activity for T lymphocytes. This study examined the role of CD13/aminopeptidase N in lymphocytic alveolitis in radiation-induced lung injury caused by a single-dose thoracic irradiation (15 Gy) in rats. Significantly increased aminopeptidase activity was detected in bronchoalveolar lavage fluid obtained from irradiated rats at 4 weeks after irradiation compared to the activity in unirradiated rats. Significantly higher aminopeptidase activity was detected on alveolar macrophages from irradiated rats at 2 and 4 weeks than on those from unirradiated rats. Western blot analysis showed an increased expression of CD13/aminopeptidase N protein in alveolar macrophages from irradiated rats at 4 weeks. Chemotactic activity for normal rat lymphocytes was detected in bronchoalveolar lavage fluid from irradiated rats at 4 weeks, and approximately 60% of the activity was inhibited by pretreatment of bronchoalveolar lavage fluid with bestatin, a specific aminopeptidase inhibitor. This study suggests that CD13/aminopeptidase N may play an important role as a lymphocyte chemoattractant in lymphocyte-mediated alveolitis in experimental radiation-induced lung injury.
Kenji Tani, Takeshi Endo and Saburo Sone : The Significance of CD13/Aminopeptidase N in Interstitial Lung Diseases, The Journal of the Japan Society for Respiratory Endoscopy, Vol.23, No.8, 731-734, 2001.
(要約)
CD13/aminopeptidase N (E.C.3.4.11.2) is an ectoenzyme located in the outer cell membrane in a variety of cells. Since CD13/aminopeptidase N was shown to induce in vitro chemotactic migration of human lymphocytes, we examined here the significance of CD13/aminopeptidase N in pulmonary sarcoidosis and radiation pneumonitis caused by a single-dose thoracic irradiation (15 Gy) in a rat model. The activity of aminopeptidase in bronchoalveolar lavage fluid (BALF) was significantly higher in sarcoidosis patients than in normal volunteers (NV) and control patients (CP). CD13/aminopeptidase N protein was detectable in alveolar macrophages (AN) from sarcoidosis patients at higher levels than in those from NV. Higher chemotactic activity for lymphocytes was detected in the BALF from sarcoidosis patients that in that from NV, and the activity was significantly decreased by the treatment with bestatin, an specific inhibitor for aminopeptidase N. Significantly increased CD13/aminopeptidase N activity and expression were also detected in BALF and AM obtained from irradiated rats at 4 weeks after irradiation compared with the activity in unirradiated rats. Chemotactic activity for normak rat lymphocytes was detected in BALF from irradiated rats at 4 weeks, and approximately 60% of the activity was inhibited by pretreatment of BALF with bestatin. This study suggests that CD13/aminopeptidase N may play an important role as a lymphocyte chemoattractant in lymphocyte-mediated alveolitis in interstitial lung diseases.
Y. Suzuki, Hiroaki Yanagawa, Yasuhiko Nishioka, N. Nishimura and Saburo Sone : Effect generation of dendritic cells from alveolar and pleural macrophages as well as blood monocytes with lung cancer, Lung Cancer, Vol.34, No.2, 195-205, 2001.
(要約)
In this study, we investigated the generation of dendritic cells (DCs) from blood monocytes and mature macrophages from untreated primary lung cancer patients. Blood monocytes were separated by adherence from blood mononuclear cells (MNC) from ten lung cancer patients and ten control subjects, and cultured for 7 days in medium with granulocyte/macrophage colony-stimulating factor (GM-CSF) plus interleukin (IL-) 4. In all cases examined, DCs with typical characteristics were obtained even in lung cancer patients after 7 days culture with these cytokines, and there was no significant difference in phenotype and stimulatory activity in allogeneic lymphocyte proliferation between DCs derived from monocytes from lung cancer patients and those from control subjects. Next, we examined whether alveolar and pleural macrophages in malignant pleural effusion separated by magnetic beads could differentiate to immunostimulatory DCs. Conventional culture conditions with GM-CSF and IL-4 did not induce efficient numbers of DCs from mature macrophages, whereas the addition of tumor necrosis factor-alpha (TNF-alpha) to GM-CSF and IL-4 effectively contributed to generate DCs. These findings suggest that both mature macrophages and blood monocytes from lung cancer patients could differentiate to DCs, and might be a useful source of DCs for immunotherapy.
Kayoko Hase, Kenji Tani, Kouji Matsushima and Saburo Sone : Increased CCR4 expression in active systemic lupus erythematosus, Journal of Leukocyte Biology, Vol.70, No.5, 749-755, 2001.
(要約)
CC chemokine receptor (CCR)4 is selectively expressed on Th2-type T cells and has been shown to be responsible for Th2-dominant immune responses. In this study, we analyzed the expression of CCR4 in active systemic lupus erythematosus (SLE) patients by FACS analysis using anti-human CCR4 monoclonal antibody and determined the clinical relevance in this disease. Higher expression of CCR4 was found on peripheral blood CD4+ T lymphocytes of active SLE patients than was found with healthy controls and inactive SLE patients. The CCR4 expression significantly correlated with the SLE disease activity index (SLEDAI) scores. The expression was dramatically decreased after the corticosteroid therapy in parallel with a serum level of double-stranded DNA antibody and SLEDAI scores. Moreover, we found that serum levels of IL-10 were increased in active SLE patients and significantly correlated with the CCR4 expression. This study suggests that Th2 immune response is predominant in the active state of SLE, and CCR4 may have relevance in regard to the disease course in SLE patients.
Masaki Hanibuchi, Seiji Yano, Yasuhiko Nishioka, Hiroaki Yanagawa, Toyokazu Miki and Saburo Sone : Immunological circumvention of multiple organ metastases of multidrug resistant human small cell lung cancer cells by mouse-human chimeric anti-ganglioside GM2 antibody KM966., Clinical & Experimental Metastasis, Vol.18, No.5, 353-360, 2001.
(要約)
serum against SBC-3/DOX cells to a similar extent compared with parental SBC-3 cells. Pretreatment of human effector cells with various cytokines induced further enhancement of the KM966-dependent ADCC against SBC-3/DOX cells. Intravenous injection of SBC-3 or SBC-3/DOX cells into natural killer (NK) cell-depleted severe combined immunodeficient (SCID) mice developed metastases in multiple organs (liver, kidneys and lymph nodes). Interestingly, SBC-3/DOX cells produced metastases more rapidly than SBC-3 cells, suggesting more aggressive phenotype of SBC-3/DOX cells than their parental cells in vivo. Systemic treatment with KM966, given on days 2 and 7, drastically inhibited the formation of multiple-organ metastases produced by both SBC-3 and SBC-3/DOX cells, indicating that KM966 can eradicate metastasis by SCLC cells irrespective of MDR phenotype. These findings suggest that the mouse-human chimeric KM966 targets the GM2 antigen, and might be useful for the immunological circumvention of multiple-organ metastases of refractory SCLC.
(キーワード)
Animals / Antibodies, Monoclonal / Antineoplastic Agents / Carcinoma, Small Cell / Cytokines / Doxorubicin / Drug Resistance, Multiple / Drug Resistance, Neoplasm / G(M2) Ganglioside / Humans / Lung Neoplasms / Male / Mice / Mice, SCID
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11467766
Yasuhiko Nishioka, Naoki Nishimura, Yoshihiro Suzuki and Saburo Sone : Human monocyte-derived and CD83(+) blood dendritic cells enhance NK cell-mediated cytotoxicity., European Journal of Immunology, Vol.31, No.9, 2633-2641, 2001.
(要約)
Dendritic cells (DC) are known to be the most potent APC and to stimulate antigen-specific T cell responses. Recently it was reported that murine DC were also capable of modulating the innate immunity by stimulating NK cells through cell-to-cell contact. In the present study, we examined whether human DC could affect NK activity. Both monocyte-derived and CD83(+) blood DC were tested. The addition of DC to cultures of CD56(+) cells resulted in the significant dose-dependent enhancement of the killing activity against various NK-sensitive targets. The resultant activity was comparable to that induced by optimal concentrations of various cytokines, including IL-2, IL-12, IL-15 and IFN-gamma. Interestingly, DC enhanced the cytotoxicity of CD3(-)CD56(+) NK cells, but not that of CD3(+)CD56(+) T cells. Experiments using transwells clearly demonstrated that the enhancement of NK activity by DC was mediated by soluble factors produced by DC. The culture supernatants of DC also stimulated NK activity. The treatment of both DC and their supernatants with anti-human IL-12 or IL-18 antibodies did not block the enhancement of NK cell-mediated cytolysis by DC, indicating that other factor(s) produced by DC were responsible for the enhancement of NK activity. These results suggest that human myeloid DC can modulate innate immunity by enhancing NK activity.
Kenji Tani, Fumitaka Ogushi, Teruki Shimizu and Saburo Sone : Protease-induced leukocyte chemotaxis and activation: roles in host defense and inflammation, The Journal of Medical Investigation : JMI, Vol.48, No.3-4, 133-141, 2001.
(要約)
The migration of leukocytes such as neutrophils, monocytes and lymphocytes into inflamed lesions is one of the critical events of inflammation. Although the traditional function of neutrophil-derived antimicrobial proteases is to ingest and kill bacteria, some neutrophil serine proteases have been shown to induce leukocyte migration and activation. Mast cell-derived chymase also has the chemotactic activity for leukocytes. During the acute phase of inflammatory and allergic diseases, the predominantly migrated cells are neutrophils and mast cells, respectively, and in the subsequent chronic phase, monocytes and lymphocytes are mainly migrated. The chemotactic activity for monocytes and lymphocytes of neutrophil-derived serine proteases and mast cell-derived chymase may have a role in switching acute inflammation to chronic inflammation and delayed-type hypersensitivity. Recently, aminopeptidase N and endothelin were shown to induce chemotactic migration of leukocytes. Thus, protease-induced leukocyte chemotaxis and activation may play an important role in immunologic events of inflammatory and allergic diseases.
Fumitaka Ogushi, Kenji Tani, Takeshi Endo, Hiroya Tada, Tetsuya Kawano, Toru Asano, Luping Huang, Yasukazu Ohmoto, Masahiro Muraguchi, Hiroki Moriguchi and Saburo Sone : Autoantibodies to IL-1 alpha in sera from rapidly progressive idiopathic pulmonary fibrosis, The Journal of Medical Investigation : JMI, Vol.48, No.3,4, 181-189, 2001.
(要約)
To clarify the clinical significance of autoantibodies to interleukin-1 alpha (IL-1 alpha autoantibodies) in rapidly progressive idiopathic pulmonary fibrosis (IPF), we measured the level of IL-1 alpha autoantibodies in serum of 11 patients on the first hospital day, when patients were admitted due to severe symptoms, and on the 21st hospital day. IL-1 alpha autoantibodies in serum were measured using radioimmunoassay, and the limitation of this assay for IL-1 alpha autoantibodies was 5 ng/ml. These antibodies were detected in 5 of 11 patients on the first hospital day. On the 21st hospital day, these antibodies were detected in all patients, and its level was increased compared with that on the first hospital day. IL-1 alpha autoantibodies that appeared in patients corresponded to that of IgG. The half life of exogenous autoantibodies was investigated following administration of autoantibody rich plasma obtained from healthy blood donors to 6 control patients (CP) and 6 progressive IPF patients. These autoantibody levels in their serum were less than 5 ng/ml before administration. Serum was obtained at the indicated time after administration of IL-1 alpha autoantibodies and the level of these autoantibodies in serum was measured, then the half life was calculated. Half life of exogenous IL-1 alpha autoantibodies in progressive IPF patients was significantly shorter than that in CP (71.3 +/- 31.8 hr vs 352.0 +/- 98.3 hr, p < 0.01). These findings suggested that IL-1 alpha autoantibodies were generated in response to the inflammatory process of rapidly progressive IPF and may act as a regulatory factor for IL-1 alpha.
Ping Cui, Kenji Tani, Hiroko Kitamura, Yuushi Okumura, Mihiro Yano, Daisuke Inui, Toshiaki Tamaki, Saburo Sone and Hiroshi Kido : A novel bioactive 31-amino acid endothelin-1 is a potent chemotactic peptide for human neutrophils and monocytes, Journal of Leukocyte Biology, Vol.70, No.2, 306-312, 2001.
(要約)
Endothelin (ET)-1(1-31) is a novel 31-amino acid-length peptide derived from big ET-1 by chymase or other chymotrypsin-type proteases and is a major ET derivative in human neutrophils. In this study, we revealed that ET-1(1-31), but not big ET, exhibited chemotactic activities toward human neutrophils and monocytes as an inflammatory mediator, although the effects were less potent than those of formyl-methionyl-leucyl-phenylalanine or interleukin-8. However, the chemotactic effects of ET-1(1-31) were much greater than those of the 21-amino acid ET-1, ET-1(1-21). Checkerboard analyses revealed that the effects are chemotactic rather than chemokinetic. The effects of ET-1(1-31) are not mediated by interleukin-8 or monocyte chemoattractant protein-1. The chemotactic effects and an increase in intracellular-free Ca(2)(+) caused by ET-1(1-31) were significantly inhibited by BQ123, an ET(A) receptor antagonist, but not by BQ788, an ET(B) receptor antagonist, suggesting that ET-1(1-31) mediates chemotaxis through an ET(A) or ET(A)-like receptor.
Tsutomu Shinohara, Naoki Nishimura, Masaki Hanibuchi, Hiroshi Nokihara, Toyokazu Miki, Hirofumi Hamada and Saburo Sone : Transduction of KAI1/CD82 cDNA promotes hematogenous spread of human lung-cancer cells in natural killer cell-depleted SCID mice, International Journal of Cancer, Vol.94, No.1, 16-23, 2001.
(要約)
KAI1, which is identical to CD82, was initially identified as a metastasis-suppressor gene for human prostate cancer, and its expression is reported to be a favorable prognostic factor for operable human lung cancer. In this study, we examined the functional role of KAI1/CD82 in the late phase of metastatic spread of human lung-cancer cells. For this, KAI1/CD82 cDNA was introduced into KAI1/CD82 low-expressing human lung-cancer cell lines, SBC-3 and PC-14, and then the metastatic potential of the transformants was analyzed by i.v. inoculation of KAI1/CD82-transduced cells, SBC-3/KAI1 and PC-14/KAI1, into NK cell-depleted SCID mice. Contrary to our expectations, KAI1/CD82 gene transfer promoted multiorgan metastasis of i.v.-inoculated human lung-cancer cells, while s.c. tumor growth was unaffected. Cancer cells from metastatic tumors of NK cell-depleted SCID mice injected i.v. with SBC-3/KAI1 expressed appreciable cell-surface KAI1/CD82, and cells not expressing KAI1/CD82 (revertants) were not detected in the tumors. Our findings indicate that under conditions where the host's natural cytotoxicity is suppressed, KAI1/CD82 may enhance the formation of tumors by circulating lung-cancer cells at metastatic sites.
Naoki Nishimura, Yasuhiko Nishioka, Tsutomu Shinohara, Hirohisa Ogawa, Sayaka Yamamoto, Kenji Tani and Saburo Sone : Novel centrifugal method for simple and highly efficient adenovirus-mediated green fluorescence protein gene transduction into human monocyte-derived dendritic cells, Journal of Immunological Methods, Vol.253, No.1-2, 113-124, 2001.
(要約)
Dendritic cells (DC) are professional antigen-presenting cells in the immune system. Gene transduction of DC with tumor-associated antigen (TAA) or other genes that enhance the immune reaction has been considered theoretically useful for DC-based immunotherapy. However, gene transduction of DC generated from human peripheral blood monocytes has been difficult due to its low efficiency, even when adenoviral vector was used at high multiplicity of infection (MOI). In the present study, we examined the effect of centrifugal force to enhance efficiency of adenovirus-mediated gene transduction into human monocyte-derived DC at various rotor speeds at various temperatures for various times. We judged the transduction efficiency using enhanced green fluorescence protein (EGFP)-expressing adenoviral vector, and the best condition for centrifugal transduction was determined as 2000 x g at 37 degrees C for 2 h at an MOI of 10 or greater. At an MOI of 50 without centrifugation, the gene transduction efficiency was about 66% and mean fluorescence intensity (MFI) of EGFP expression was about 150 (at 37 degrees C for 2 h). With centrifugal transduction (2000 x g at an MOI of 50 at 37 degrees C for 2 h), 86% or more DC were gene-modified, and especially, MFI of EGFP expression was highly enhanced (MFI: about 3100 or greater). Centrifugally gene-transduced DC were not damaged and were thoroughly functional as measured by mixed lymphocyte reaction (MLR). The centrifugal method was also applicable to human monocytes and K562 cells. The centrifugal transduction method with adenoviral vector might be helpful for the generation of gene-modified DC.
H Nokihara, Seiji Yano, Yasuhiko Nishioka, M Hanibuchi, T Higasida, T Tsuruo and Saburo Sone : A new quinoline derivative MS-209 reverse multidrug resistance and inhibits multiorgan metastases by P-glycoprotein-expressing human small cell lung cancer cells, Japanese Journal of Cancer Research, Vol.92, No.7, 785-792, 2001.
(要約)
Development of distant metastases and acquired multidrug resistance (MDR) are major problems in therapy for human small cell lung cancer (SCLC). MS-209 is a novel quinoline compound, which reverses P-glycoprotein (P-gp)-mediated MDR. We previously reported that MS-209 reversed in vitro MDR of human SCLC (SBC-3 / ADM and H69 / VP) cells expressing P-gp. In the present study, we determined the therapeutic effect of MS-209 in combination with chemotherapy against multiorgan metastases of MDR SCLC cells. SBC-3 / ADM cells expressing P-gp were highly resistant to etoposide (VP-16), adriamycin (ADM), and vincristine (VCR) in vitro, compared with parental SBC-3 cells lacking P-gp expression. MS-209 restored chemosensitivity of SBC-3 / ADM cells to VP-16, ADM, and VCR in a dose-dependent manner in vitro. Intravenous injection with SBC-3 or SBC-3 / ADM cells produced metastatic colonies in the liver, kidneys and lymph nodes in natural killer (NK) cell-depleted severe combined immunodeficiency (SCID) mice, though SBC-3 / ADM cells more rapidly produced metastases than did SBC-3 cells. Treatment with VP-16 and ADM reduced metastasis formation by SBC-3 cells, whereas the same treatment did not affect metastasis by SBC-3 / ADM cells. Although MS-209 alone had no effect on metastasis by SBC-3 or SBC-3 / ADM cells, combined use of MS-209 with VP-16 or ADM resulted in marked inhibition of metastasis formation by SBC-3 / ADM cells to multiple organs. These findings suggest that MS-209 reversed the MDR of SBC-3 / ADM cells, but not SBC-3 cells, growing in the various organs, and inhibited metastasis formation in vivo. Therefore, this chemosensitizing agent, MS-209, may be useful for treatment of refractory SCLC patients with multiorgan metastases.
Huang Luping, Fumitaka Ogushi, Kenji Tani, Hirohisa Ogawa, Tetsuya Kawano, Takeshi Endo, Keisuke Izumi, Nobuya Sano, Junji Ueno, Hiromu Nishitani and Saburo Sone : Thrombin Promotes Fibroblast Proliferation during the Early Stages of Experimental Radiation Pneumonitis, Radiation Research, Vol.156, No.1, 45-52, 2001.
(要約)
Huang, L., Ogushi, F., Tani, K., Ogawa, H., Kawano, T., Endo, T., Izumi, K., Ueno, J., Nishitani, H. and Sone, S. Thrombin Promotes Fibroblast Proliferation during the Early Stages of Experimental Radiation Pneumonitis. Radiat. Res. 156, 45-52 (2001). To clarify the role of thrombin in the pathogenesis of radiation-induced pneumonitis, we measured the thrombin activity and fibroblast growth-inducing activity in bronchoalveolar lavage fluid obtained from the irradiated lungs of rats at 1, 2, 4, 8 and 18 weeks after irradiation. Thrombin activity was not detected in the bronchoalveolar lavage fluid from unirradiated rats, but the bronchoalveolar lavage fluid from irradiated rats showed significantly increased thrombin activity which reached a maximum at 4 weeks after treatment. Higher fibroblast growth-inducing activity was detected in the bronchoalveolar lavage fluid from irradiated rats at 4 and 18 weeks than in fluid from unirradiated rats. Bronchoalveolar lavage fluid from irradiated rats that were pretreated with the thrombin inhibitors antithrombin III and argatroban showed significantly inhibited fibroblast growth-inducing activity and thrombin activity at 4 weeks. However, these thrombin inhibitors did not inhibit fibroblast growth-inducing activity in bronchoalveolar lavage fluid from irradiated rats at 18 weeks. Purified rat thrombin similarly induced proliferation of fibroblasts derived from irradiated and unirradiated rats. These findings suggest that thrombin may play an important role as a fibroblast growth-inducing factor during the early stages of radiation pneumonitis.
Prahlad Parajuli, Hiroaki Yanagawa, Masaki Hanibuchi, Eiji Takeuchi, Toyokazu Miki, Seiji Yano and Saburo Sone : Humanized anti-ganglioside GM2 antibody is effective to induce antibody-dependent cell-mediated cytotoxicity in mononuclear cells from lung cancer patirnts, Cancer Letters, Vol.165, No.2, 179-184, 2001.
(要約)
Ganglioside GM2 is one of the major gangliosides expressed on the cell surface of human tumors including lung cancer. We have previously reported that a mouse-human chimeric monoclonal antibody (mAb), KM966, against GM2 promotes the lysis of lung cancer cells by human blood mononuclear cells (MNC) of healthy donors. In this study, we examined antibody-dependent cell-mediated cytotoxicity (ADCC) of MNC, using KM966 mAb and its humanized counterpart, KM8969, in 16 lung cancer patients and 18 control patients. The ADCC activity was assessed by 4-h (51)Cr release from GM2 positive SBC-3 small cell lung cancer cells. MNC from lung cancer patients exhibited similar ADCC activity to those from control patients when KM966 and KM8969 were used as mAb. Moreover, effective ADCC activity was observed even in MNC from advanced lung cancer patients. These observations suggest the potential activity of humanized anti-GM2 mAb (KM8969), as well as chimeric KM966, in biological therapy for lung cancer patients.
Naoki Nishimura, Yasuhiko Nishioka, Tsutomu Shinohara and Saburo Sone : Enhanced efficiency by centrifugal manipulation of adenovirus-mediated interleukin 12 gene transduction into human monocyte-derived dendritic cells., Human Gene Therapy, Vol.12, No.4, 333-346, 2001.
(要約)
Transduction of dendritic cells (DCs) with genes encoding tumor-associated antigen or with other genes that enhance immune reaction has been theorized to be potentially useful for enhancing the efficiency of DC-based immunotherapy. However, gene transduction of DCs generated from human peripheral blood monocytes has been of limited use because of the low efficiency. Here, we report that the efficiency of in vitro adenovirus-mediated gene transduction into human monocyte-derived DCs can be dramatically enhanced by centrifugation. The best conditions for centrifugal gene transduction were determined to be as follows: 2000 x g at 37 degrees C for 2 hr at a multiplicity of infection (MOI) of 10 or greater. By this centrifugal method, approximately 88 and 70% of DCs were gene transducible at an MOI of 50 and 10, respectively. Functional analysis showed that DCs transduced with human interleukin 12 (IL-12)-expressing adenoviral vector under the optimal conditions of centrifugation stably produced IL-12 protein at high levels (8.1 ng/10(6) cells/48 hr). IL-12 gene-modified DCs (DC/IL-12) displayed a more mature phenotype than nontransduced DCs, as judged by decreased expression of CD1a and increased expression of CD83, B7.1 (CD80), B7.2 (CD86), and MHC class I and II molecules. DC/IL-12 showed a high phagocytic ability similar to nontransduced DCs and were significantly superior to control DCs in the stimulation of autologous and allogeneic T lymphocyte responses. The centrifugal transduction method with adenoviral vector might be useful for efficient generation of gene-modified DCs because it is very simple, highly efficient, reproducible, and not cytopathic. IL-12 gene-modified human DCs may be therapeutically useful as a good adjuvant in DC-based immunotherapy.
Saburo Sone, Tsutomu Shinohara, Yasuhiko Nishioka and Seiji Yano : Symposium on molecular pathogenesis of respiratory diseases and its clinical implication. 4. Molecular pathogenesis of lung cancer and its molecular targeted therapy, Internal Medicine, Vol.40, No.2, 167-170, 2001.
Haruko Tanaka, Kenichi Maeda, Yoichi Nakamura, Masahiko Azuma, Hiroaki Yanagawa and Saburo Sone : CD40 and IFN-γ dependent T cell activation by human bronchial epithelial cells, The Journal of Medical Investigation : JMI, Vol.48, No.1, 2, 109-117, 2001.
(要約)
We examined whether freshly isolated human bronchial cells (HBEC) and bronchial epithelial cell line/BEAS-2B cells expressed surface molecules required for APC function. These cells expressed CD40 and ICAM-1, but not B7-1, B7-2 or HLA-DR molecules. Treatment of these cells with IFN-gamma resulted in enhanced expression of CD40 and ICAM-1 as well as induction of HLA-DR expression. Th2 cytokines such as IL-4 and IL-5, proinflammatory cytokine of GM-CSF and nonspecific activator endotoxin had no effect on these phenotypic expressions. Functional examinations showed that allogeneic lymphocytes purified from peripheral blood strongly proliferated in response to BEAS-2B cells cultured with IFN-gamma, but only weakly compared with those without IFN-gamma. When allogeneic lymphocytes were purified to CD4+ cells, the proliferative response against BEAS-2B cells was abolished. Blockade of CD40-CD40L interaction by anti-CD40 antibody also inhibited the proliferation of lymphocytes to BEAS-2B cells, although this treatment showed a minimum effect on the response to allogeneic MNC. Thus, bronchial epithelial cells have the ability to present allogeneic antigens to T cells in both CD40- and IFN-gamma-dependent manners under the presence of third party cells that transduce co-stimulatory signals.
Tsutomu Shinohara, Toyokazu Miki, Naoki Nishimura, Hiroshi Nokihara, Hirofumi Hamada, Naofumi Mukaida and Saburo Sone : Nuclear Factor-kB-dependent Expression of Metastasis Suppressor KAI1/CD82 Gene in Lung Cancer Cell Lines Expressing Mutant p531, Cancer Research, Vol.61, No.2, 673-678, 2001.
(要約)
KAI1/CD82 has been shown to be a metastasis suppressor for several human cancers, and a recent study revealed that wild-type tumor suppressor p53 can directly activate KAI1/CD82 gene expression. However, the response of KAI1/CD82 expression in cancer cells to exogenous stimulants has not been investigated. The present study examined whether tumor necrosis factor (TNF), which mediates many of the cellular responses associated with inflammatory reactions or cancer progression, can affect the KAI1/CD82 expression in lung cancer cells and, if so, whether nuclear factor (NF)-kappaB, a key molecule in TNF-mediated gene expression, is involved in the mechanism of KAI1/CD82 induction. Our results demonstrated that expression of KAI1/CD82 in PC-14 cells expressing mutant p53 could be augmented by TNF-alpha, and that transfer of the gene for a specific inhibitor of NF-kappaB, IkappaB alphaSR (mutant IkappaB alpha; NF-kappaB super-repressor), into PC-14 cells could inhibit this augmentation. The amount of NF-kappaB in the nucleus of PC-14/IkappaB alphaSR cells correlated well with KAI1/CD82 mRNA and protein expression. In addition, IkappaB alphaSR gene transfer inhibited the spontaneous expression of KAI1/CD82 protein in KAI1/CD82-high-expressing RERF-LC-OK cells, which contain a mutant-type p53. These observations indicate that NF-kappaB activation may play a role in the regulation of KAI1/CD82 expression in lung cancer cells independently of wild-type p53, and suggest that KAI1/CD82 expression may be regulated by interaction with the host microenvironment.
Eiji Takeuchi, Hiroaki Yanagawa, Yoshihiro Suzuki, Kunihiro Shinkawa, Hiroyasu Bandoh and Saburo Sone : INTERLEUKIN (IL-)15 HAS LESS ACTIVITY THAN IL-2 TO PROMOTE TYPE 2 CYTOKINE PREDOMINANCE IN TUMOUR-ASSOCIATED MONONUCLEAR CELLS FROM LUNG CANCER PATIENTS, Cytokine, Vol.13, No.2, 119-123, 2001.
Hiroshi Nokihara, Hiroaki Yanagawa, Yasuhiko Nishioka, Seiji Yano, Naofumi Mukaida, Kouji Matsushima and Saburo Sone : Natural killer cell-dependent suppression of systemic spread of human lung adenocarcinoma cells by monocyte chemoattractant protein-1 gene transfection in severe combined immunodeficient mice., Cancer Research, Vol.60, No.24, 7002-7007, 2000.
119.
Naoki Nishimura, Yasuhiko Nishioka, Tsutomu Shinohara, Kenji Tani and Saburo Sone : Down-regulation by a new anti-inflammatory compound, FR167653, of differentiation and maturation of human monocytes and bone marrow CD34(+) cells to dendritic cells, International Journal of Immunopharmacology, Vol.22, No.7, 501-514, 2000.
120.
Kenya Sumitomo, Akira Kurisaki, Norio Yamakawa, Kunihiro Tsuchida, Eiji Shimizu, Saburo Sone and Hiromu Sugino : Expression of a TGF-β1 inducible gene, TSC-36, causes growth inhibition in human lung cancer cell lines, Cancer Letters, Vol.155, No.1, 37-46, 2000.
121.
Kenji Tani, Fumitaka Ogushi, Hiroshi Kido, Tetsuya Kawano, Yuichi Kunori, Takashi Kamimura, Ping Cui and Saburo Sone : Chymase is a potent chemoattractant for human monocytes and neutrophils, Journal of Leukocyte Biology, Vol.67, No.4, 585-589, 2000.
(要約)
Chymase is a major chymotrypsin-like serine protease expressed in the secretory granules of mast cells in many mammalian species. In this study, we revealed the chemotactic activity of chymase for human mononuclear cells and neutrophils with a 48-well microchemotaxis chamber technique. Human chymase showed the potent chemotactic activity for monocytes and neutrophils dose-dependently in a concentration range from 0.1 to 10 microg/mL, corresponding to about 4-400 microM. The activity was as potent as that of N-formyl-methionyl-leucyl-phenylalanine. Chymase also stimulated cell migration of lymphocytes and purified T cells, but checkerboard analysis revealed that the effect was chemokinetic rather than chemotactic. Inhibition of chymase activities with chymase inhibitors, such as antileukoprotease and Bowman-Birk soybean trypsin inhibitor, significantly inhibited the chemotactic activity of chymase, suggesting that the proteolytic activity of chymase participates in the chemotactic activity. Our results suggest that mast cell chymase acts as a chemoattractant, and may play a role in the accumulation of inflammatory cells in development of the chronic inflammatory responses of allergic and nonallergic diseases.
Hitoe Torisu, Mayumi Ono, Hiromaro Kiryu, Masutaka Furue, Yasukazu Ohmoto, Juichiro Nakayama, Yasuhiko Nishioka, Saburo Sone and Michihiko Kuwano : Macrophage infiltration correlates with tumor stage and angiogenesis in human malignant melanoma: Possible involvement of TNFα and IL-1α, International Journal of Cancer, Vol.85, No.2, 182-188, 2000.
(要約)
We examined whether macrophage infiltration is associated with angiogenesis in cutaneous melanoma. The numbers of macrophages and microvessels increased significantly with increasing depth of tumor and with tumor angiogenesis. Macrophage infiltration thus appeared to provide a useful diagnostic marker for the progression of cutaneous melanoma. We further examined whether human melanoma cells produce angiogenic factors in response to macrophage-derived cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin-1 alpha (IL-1alpha). Treatment of melanoma cells with TNFalpha and IL-1alpha in vitro enhanced the production of interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF), and of basic fibroblast growth factor (bFGF) to a lesser degree, in human melanoma cells. Lipopolysaccharide (LPS)-activated human monocytes enhanced production of IL-8, VEGF, TNF alpha, as well as IL-1alpha, but not bFGF. Co-culture of human monocytes and human melanoma cells was also found to significantly enhance production of IL-8 and VEGF in the absence and presence of LPS, compared with either monocytes or melanoma cells alone. The production of IL-8 and VEGF from co-cultured melanoma cells and LPS-activated monocytes was blocked when anti-TNF-alpha antibody or anti-IL-1alpha antibody was co-administrated. This is direct evidence that production of the potent angiogenic factors IL-8 and VEGF from melanoma cells is up-regulated through TNFalpha and/or IL-1alpha secreted by activated monocytes/macrophages, influencing both tumor growth and angiogenesis in melanomas.
Singh Mahendra Sukh, Hiroaki Yanagawa, Masaki Hanibuchi, Toyokazu Miki, Okamura Haruki and Saburo Sone : Augmentation by interleukin-18 of MHC-nonrestricted killer activity of human peripheral blood mononuclear cells in response to interleukin-12, International Journal of Immunopharmacology, Vol.22, No.1, 35-43, 2000.
(要約)
Interleukin (IL)-18 is a novel cytokine with pleiotropic functions. In the present study, we examined the induction of the killer activity of peripheral blood mononuclear cells (MNC) against lung cancer cell lines upon treatment with IL-18 in combination with IL-12. Cytotoxic activity was measured by standard (51)Cr release assay. IL-18 (100 ng/ml) was found to significantly augment IL-12-induced killer activity in a MHC-nonrestricted manner against allogeneic NK-resistant Daudi cells and lung cancer cell lines: SBC-3, RERF-LC-AI and A549. IL-18 could augment IL-12-induced killer activity both at the optimal as well as suboptimal doses of the latter. However, IL-18 was found to have little effect on the killer activity of MNC induced by optimal or suboptimal dose of IL-2 or IL-15. Treatment of MNC with IL-18 in combination with IL-12 for a period of more than 4 days was observed to optimally induce the killer activity. As for induction of IFN-gamma production by MNC, IL-18 augmented that induced by IL-2 and IL-15, as well as that induced by IL-12. These results show the potential of IL-18 in combination with IL-12 for clinical application in treatment of cancer.
Toyokazu Miki, Seiji Yano, Masaki Hanibuchi and Saburo Sone : Bone metastasis model with multi-organ dissemination of human small cell lung cancer (SBC-5) cells in nutural killer-cell depleted SCID mice, Oncology Research, Vol.12, No.5, 209-217, 2000.
(要約)
Lung cancer is commonly associated with multiorgan metastasis, and bone is a frequent metastatic site for lung cancer. Nevertheless, no bone metastasis model of lung cancer with multiorgan dissemination is available, which could provide opportunity to study the molecular pathogenesis. We examined the abilities of eight human lung cancer cell lines injected intravenously into natural killer (NK) cell-depleted SCID mice to generate metastatic nodules in bone and multiple organs, and explored the correlation of the parathyroid hormone-related protein (PTHrP) with the bone metastasis. Although all the small-cell carcinoma cell lines (SBC-5, SBC-3, SBC-3/ADM, H69, H69/VP) formed metastatic nodules in multiple organs (liver, kidney, and lymph nodes), only SBC-5 cells reproducibly developed bone metastases. Squamous cell carcinoma (RERF-LC-AI) cells metastasized mainly into the liver and kidneys, whereas adenocarcinoma (PC-14, A549) mainly produced colonies in the lungs. As assessed by X-ray photography, the osteolytic bone metastases produced by SBC-5 cells were detected as early as on day 28, and all recipient mice developed bone metastasis by day 35. The expression of PTHrP in eight cell lines was directly correlated with the formation of bone metastasis. No correlation was observed between the formation of bone metastasis and the expression of other metastasis-related cytokines (IL-1, IL-6, IL-8, IL-10, IL-11, TNF-alpha, VEGF, M-CSF). Consistent with the formation of bone metastasis by SBC-5 cells, the levels of PTHrP and calcium in the mouse serum were increased in a time-dependent manner, suggesting that PTHrP produced by human lung cancer may play a crucial role in the formation of bone metastasis and hypercalcemia. These findings indicate that a bone metastasis model of SBC-5 cells may be useful for clarifying the molecular aspects of the metastatic processes in different organ microenvironments and the development of therapeutic modalities for lung cancer patients with bone metastases.
(キーワード)
Animals / Body Weight / Bone Neoplasms / Calcium / Carcinoma, Small Cell / Carcinoma, Squamous Cell / Cytokines / Disease Models, Animal / Humans / Hypercalcemia / Kidney Neoplasms / Killer Cells, Natural / Liver Neoplasms / Lung Neoplasms / Lymphocyte Depletion / Male / Mice / Mice, SCID / Parathyroid Hormone-Related Protein / Proteins / Severe Combined Immunodeficiency / Time Factors / Tumor Cells, Cultured
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11417746
K Miki, Eiji Shimuzu, Seiji Yano, Kenji Tani and Saburo Sone : Demethylation by 5-aza-2'-deoxycytidine (5-azadC) of p16INK4A gene results in downregulation of vascular endothelial growth factor expression in human lung cancer cell lines, Oncology Research, Vol.12, No.8, 335-342, 2000.
(要約)
Vascular endothelial growth factor (VEGF) plays a pivotal role in tumor progression via angiogenesis. Recently, gene transduction of wild-type p16INK4A, tumor suppressor gene, has been shown to result in downregulation of VEGF expression in p16INK4A-deleted glioma cells. Because expression of p16INK4A is regulated by methylation of the p16INK4A gene, we examined whether demethylation of the p16INK4A gene by 5-aza-2'-deoxycytidine (5-azadC) could cause the protein expression of VEGF as well as of p16INK4A in human lung cancer cells. For this, five different lung cancer cell lines with or without loss of p16 activity were used. H841 and Ma-10 cells had the methylated p16INK4A gene without expression of p16INK4A protein, whereas Ma-1 and H209 cells had the unmethylated p16INK4A gene with constitutive expression of p16INK4A protein. Neither the p16INK4A gene nor p16INK4A protein was detected in A549 cells. Treatment with 5-azadC caused demethylation of the p16INK4A gene with reexpression of p16INK4A protein in H841 and Ma-10 (methylated p16INK4A gene dominant) cell, but not in other cell lines such as Ma-1, H209 (unmethylated p16INK4A gene dominant), or A549 (p16INK4A gene deleted). In a parallel experiment, 5-azadC inhibited production of VEGF protein by H841 and Ma-10 cells, especially in the later hypermethylated cells, but not Ma-1, H209, or A549 cells. RT-PCR analysis showed that Ma-10 cells expressed VEGF isoforms 121, 165, and 189, all of which were inhibited by 5-azadC. These findings indicate that the methylation status of the p16INK4A gene plays an important role in the regulation of angiogenesis associated with progression of lung cancer, through regulation of VEGF expression.
Takahashi Toshiya, Kenichi Maeda, Yoichi Nakamura, Yoshio Okano, N. Ge and Saburo Sone : Interleukin-10 inhibits the production of inflammatory cytokines by antigen-stimulated mononuclear cells from asthmatic patients, Allergology International, Vol.49, No.1, 55-62, 2000.
Masaki Hanibuchi, Yasuhiko Nishioka, Hiroaki Yanagawa, Seiji Yano, Parajuli Prahlad, Bando Masashi and Saburo Sone : Human interferon-gamma enhances expression of ganglioside GM2 on human lung cancer cells and their susceptibility for antiganglioside GM2 monoclonal antibody-dependent cellular cytotoxicity., Oncology Research, Vol.12, No.4, 173-179, 2000.
(要約)
Interferons are known to modulate several cellular functions by the induction of various proteins. In this study, we demonstrated that human interferon-gamma (HuIFN-gamma) enhanced the expression of ganglioside GM2 (GM2), which is a kind of tumor-associated antigen substantially expressed in human lung cancer and that human lung cancer cells expressing GM2 became more susceptible to anti-GM2 monoclonal antibody (mAb)-dependent tumor cell killing mediated by human effector cells after HuIFN-gamma treatment. GM2 expression on human lung cancer cells treated with or without HuIFN-gamma was measured by flow cytometry. The antibody-dependent cellular cytotoxicity (ADCC) activity was assessed by 4-h 51Cr release assay. HuIFN-gamma enhanced GM2 expression on human small-cell lung cancer (SCLC), SBC-3, and human non-small-cell lung cancer (NSCLC), A549 cells in a dose-dependent manner. The optimal concentration of HulIFN-gamma was 1,000 U/ml. The effect of HulFN-gamma reached maximum after 4 days of culture. HulFN-gamma did not have any effect to enhance the expression of other gangliosides in SBC-3 cells. No other cytokines used in this study modulated GM2 expression in SBC-3 cells. Anti-GM2 mAb-dependent ADCC activities induced by lymphocytes and monocytes were more potent against IFN-gamma-treated SBC-3 and A549 cells than nontreated cells. Taken together, HulFN-gamma combined with anti-GM2 mAb may be useful for immunotherapy against GM2-positive human lung cancer.
Nakamura Kazuyasu, Masaki Hanibuchi, Seiji Yano, Tanaka Yuko, Fujino Ikuko, Inoue Miho, Takezawa Toshiaki, Shitara Kenya, Saburo Sone and Hanai Nobuo : Apoptosis induction of human lung cancer cell line in multicellular heterospheroids with humanized antiganglioside GM2 monoclonal antibody., Cancer Research, Vol.59, No.20, 5323-5330, 1999.
(要約)
The chimeric antiganglioside GM2 monoclonal antibody (MAb) KM966, which showed high effector functions such as complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity (ADCC), potently suppressed growth and metastases of GM2-positive human cancer cells inoculated into mice. To further improve the therapeutic efficacy of the anti-GM2 MAb in humans, we constructed a humanized anti-GM2 MAb, KM8969. The humanized KM8969 was more efficient in supporting ADCC against GM2-positive human cancer cell lines than the chimeric KM966, whereas complement-dependent cytotoxicity was slightly reduced in the humanized KM8969. In addition, the humanized KM8969 was shown to exert potent ADCC mediated by both lymphocytes and monocytes. To investigate the effect of the humanized KM8969 on the biological function of GM2 in the condition physiologically mimicking formation and growth of cancer masses, the heterospheroids composed of normal human dermal fibroblasts and GM2-positive human lung cancer cells were developed. Interestingly, the humanized KM8969 gave rise to growth inhibition of heterospheroids without dependence of the effector functions. Morphological and immunocytochemical analysis suggested that the inhibitory effect was due to the apoptosis of GM2-positive cancer cells in the heterospheroids. The result indicates that GM2 captured by the antibody on the cell surface loses its physiological function that plays a critical role in maintaining the three-dimensional growth of cancer cells in contact with its own cells or other type of cells in a microenvironment. The humanized KM8969, which can destroy the cancer cells via blocking functional GM2 on the cell surface as well as the effector functions, would have extraordinary potential in human cancer therapy.
Hiroaki Yanagawa, H Goto, Kouji Maniwa, Fumiitaka Ogushi, K Takahashi, Yasumasa Monden, Takanori Hirose, Nobuya Sano and Saburo Sone : A case of resectable lung adenocarcinoma associated with sarcoidosis, Medical Oncology, Vol.16, No.3, 216-220, 1999.
(要約)
A 71-year-old woman with uveitis was referred to our hospital for further examination of the possible underlying diseases. In roentgenological examination with plain X-ray and CT scan, hilar and mediastinal lymphadenopathy and a mass shadow in the right upper lung field was observed, whereas fibrotic changes were not obvious in both lung fields. Transbronchial lung biopsy with fiberoptic bronchoscope revealed granulomatous interstitial pneumonia. CD4-positive lymphocytes were increased in bronchoalveolar lavage. The patient was diagnosed as having sarcoidosis. Subsequently, right upper lobectomy was performed, and Stage I lung adenocarcinoma was diagnosed. The patient is under follow up without medication and the disease has been stable for two years. A relationship between epithelioid granulomatosis and malignant diseases is discussed and a review of the literature is given. Since it is still controversial as to the incidence of malignant diseases in sarcoidosis patients, it is important to accumulate data on these associations.
Seiji Yano, Masaki Hanibuchi, Yasuhiko Nishioka, Hiroshi Nishihara, Naoki Nishimura, Takashi Tsuruo and Saburo Sone : Combined therapy with anti-p-glycoprotein antibody and macrophage colony-stimulating factor gene transduction for multiorgan metastases of multidrug-resistant human small cell lung cancer in nk cell-depleted scid mice, International Journal of Cancer, Vol.82, No.1, 105-111, 1999.
(要約)
Our aim was to determine the antimetastatic potential of anti-P-glycoprotein (P-gp) antibodies (Abs) against multidrug-resistant (MDR) human small cell lung cancer (SCLC) cells expressing P-gp. Human SCLC cells H69 (P-gp negative) and its etoposide-resistant variant H69/YP (P-gp positive) were used. H69 and H69/VP cells injected i.v. metastasized to the liver, kidneys and systemic lymph nodes of NK cell-depleted severe combined immunodeficient (SCID) mice. H69/VP cells, but not H69 cells, were resistant to treatments with vindesine. Treatment with mouse-human chimeric anti-P-gp Ab (MH162) and its mouse counterpart (MRK-16) reduced metastasis of H69/VP cells in various organs and prolonged the survival of tumor-bearing mice, although they were less effective if injected at late times (after 28 days). Treatment with another mouse anti-Pgp Ab, MRK-17, was effective only against liver metastasis. MH162 and MRK-16 efficiently induced Ab-dependent cellular cytotoxicity (ADCC) by peritoneal macrophages against H69/VP cells in vitro, but MRK-17 was less effective, in accordance with their in vivo antimetastatic potential. Gene transfection of macrophage colony-stimulating factor (M-CSF) into H69/VP cells to augment macrophage-mediated ADCC resulted in inhibition of metastasis to the liver and lymph nodes, but not kidneys. Combined treatment with a low dose of MRK-16 completely cured metastasis of M-CSF transfectant, but not of the mock transfectant. Our findings suggest that while anti-P-gp Abs had antimetastatic potential against SCLC cells expressing P-gp, combined treatment with M-CSF gene transduction to augment the therapeutic efficacy of anti-P-gp Abs may be beneficial for eradicating metastatic MDR SCLC in humans.
Prahlad Parajuli, Yasuhiko Nishioka, Naoki Nishimura, Sukh Mahendra Singh, Masaki Hanibuchi, Hiroshi Nokihara, Hiroaki Yanagawa and Saburo Sone : Cytolysis of human dendritic cells by autologous lymphokine-activated killer cells: participation of both T cells and NK cells in the killing., Journal of Leukocyte Biology, Vol.65, No.6, 764-760, 1999.
(要約)
Dendritic cells (DC) play a key role in the initiation of immune response by stimulating the naive T cells. The fate of DC after the initiation of immune response is not clearly understood. Although there are few reports implicating natural killer (NK) cells in the elimination of DC, killing of DC by LAK cells, and specifically by T cells, has not been studied. In this study, we observed that DC, generated from monocytes, in vitro in the presence of granulocyte-macrophage colony-stimulating factor, interleukin-4 (IL-4), and tumor necrosis factor alpha were susceptible to cytolysis by lymphokine-activated killer (LAK) cells induced in the presence of IL-2 and IL-15 but not IL-12 alone. However, LAK cells induced by a combination of IL-12 and suboptimal dose of IL-2 were cytotoxic to DC. When purified lymphocytes were activated with IL-2, the CD8+/CD57- fraction (T-LAK), but not the CD8-/CD57+ fraction (NK-LAK) was cytotoxic to autologous DC. However, when unseparated peripheral blood mononuclear cells were used to generate LAK cells, both T-LAK and NK-LAK fractions showed equal cytotoxicity against autologous DC. Monoclonal antibodies against CD54, CD11a, and CD18 significantly inhibited the cytolysis, indicating that the killing involves the engagement of CD54 with its ligands.
Hiroshi Nokihara, Yasuhiko Nishioka, Seiji Yano, Naofumi Mukaida, Kouji Matsushima, Takashi Tsuruo and Saburo Sone : Monocyte chemoattractant protein-1 gene modification of multidrug-resistant human lung cancer enhances antimetastatic effect of therapy with anti-P-glycoprotein antibody in SCID mice, International Journal of Cancer, Vol.80, No.5, 773-780, 1999.
(要約)
Distant metastases and multidrug resistance are critical problems in the therapy of human small cell lung cancer (SCLC). In this study, we investigated whether transduction of the monocyte chemoattractant protein-1 (MCP-1) gene into multidrug-resistant (MDR) human lung cancer cells affected the formation of metastases or their inhibition by the anti-P-glycoprotein (P-gp) monoclonal antibody (MAb) MRK16. MDR human SCLC (H69/VP) cells were transduced with the human MCP-1 gene inserted into the expression vector BCMGSNeo. MCP-1 gene transduction had no effect on drug sensitivity, the expression of surface antigens or the in vitro proliferation of H69/VP cells. Using the metastatic model of NK cell-depleted SCID mice, H69/VP cells transduced with the MCP-1 gene were inoculated intravenously (i.v.) and formed metastatic colonies in the liver, kidneys and lymph nodes, similar to those formed by parent or mock-transduced cells. However, systemic treatment of the mice with MRK16 reduced the metastases of H69/VP cells in the liver, kidneys and lymph nodes, and was significantly more effective in inhibiting the metastases of MCP-1 producing H69/VP than those of mock-transduced cells. MCP-1 gene transduction significantly prolonged the survival of tumor-bearing mice treated with MRK16. Our findings suggest that local production of MCP-1 in the tumor site increases the anti-P-gp antibody-dependent cell-mediated cytotoxicity, and the MCP-1 gene-induced modification of MDR human SCLC cells thereby enhances the antimetastatic effect of therapy with anti-P-gp antibody. Thus, the accumulation of effector cells in the tumor site is a very important factor in the therapy using the anti-P-gp antibody.
Prahlad Parajuli, Seiji Yano, Yasuhiko Nishioka, Hiroshi Nokihara, Masaki Hanibuchi, Naoki Nishimura, Teruhiro Utsugi and Saburo Sone : Therapeutic Efficacy of a New Topoisomerase I and II Inhibitor TAS-103, Against Both P-Glycoprotein Expressing and Non-expressing Drug-Resistant Human Small-Cell Lung Cancer, Oncology Research, Vol.11, No.5, 219-224, 1999.
(要約)
We examined the effect of a novel topoisomerase I and II (topo I and II) inhibitor, TAS-103, on P-glycoprotein (P-gp)-expressing and -nonexpressing drug-resistant human small-cell lung cancer (SCLC) cells in vitro and in vivo. We observed that TAS-103 was effective in inhibiting in vitro proliferation of human SCLC (SBC-3 and H69) cells and their drug-resistant variants SBC-3/ADM or SBC-3/CDDP and H-69/VP, respectively. SBC-3/ADM and H-69/VP expressed high P-gp, whereas SBC-3/CDDP did not. TAS-103 also effectively reduced the tumor growth (more than 50% inhibition) of the parental as well as MDR SCLC cells grown SC in nude mice. Adriamycin (ADM) and cisplatin (CDDP), on the other hand, were effective only against the parental cells, while these drugs failed to inhibit the respective drug-resistant variants in vitro or in vivo. TAS-103 was observed to induce apoptosis dose dependently in the parental as well as drug-resistant SCLC cells as analyzed after 48 h of in vitro treatment, suggesting that the stabilization of cleavable topo I- or II-DNA complexes by topo I and II inhibitors like TAS-103 is followed by apoptosis of the cells. Overall, our study suggests that TAS-103 may have clinical application against drug-resistant human SCLC.
(キーワード)
ATP-Binding Cassette, Sub-Family B, Member 1 / Aminoquinolines / Animals / Antineoplastic Agents / Apoptosis / Carcinoma, Small Cell / Drug Resistance, Neoplasm / Humans / Indenes / Lung Neoplasms / Male / Mice / Mice, Inbred BALB C / Mice, Nude / Neoplasm Proteins / Topoisomerase I Inhibitors / Topoisomerase II Inhibitors / Tumor Cells, Cultured
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10608616
Saburo Sone, Seiji Yano, Masaki Hanibuchi, Hiroshi Nokihara, Naoki Nishimura, Toyokazu Miki, Yasuhiko Nishioka and Tsutomu Shinohara : Heterogeneity of multiorgan metastases of human lung cancer cells genetically engineered to produce cytokines and reversal using chimeric monoclonal antibodies in natural killer cell-depleted severe combined immunodeficient mice, Cancer Chemotherapy and Pharmacology, Vol.43, No.Supplement 1, S26-S31, 1999.
(要約)
Lung cancer is a major cause of cancer deaths, most of which can be attributed to distant multiorgan metastases. To examine the cellular and molecular mechanisms of lung cancer metastasis to distant organs, we have established novel models of human lung cancer (small cell and non-small cell lung cancer) metastasis in natural killer cell-depleted severe combined immunodeficient (SCID) mice. We investigated whether local production of the cytokines responsible for regulation of macrophage function at tumor growth sites affects the pattern of lung cancer metastasis in distant organs. Several lung cancer cell lines were genetically engineered to produce human macrophage colony-stimulating factor (M-CSF) and monocyte chemoattractant protein-1 (MCP-1), and their metastatic potentials were assessed. Interestingly, M-CSF gene transduction had an antimetastatic effect for the liver and lymph nodes, but not the kidneys. In contrast, MCP-1 gene-modified lung cancer cells and their parent cells had identical metastatic potentials. These findings indicate a possible role for cytokines and suggest that lung cancer has metastatic heterogeneity. Examining ways of controlling human lung cancer metastases, we investigated the antimetastatic effect of chimeric monoclonal antibodies (MAbs) against P-glycoprotein and ganglioside GM2 (MH162 and KM966, respectively). Both MAbs, when given on days 2 and 7, inhibited the development of distant metastases of lung cancer in a dose-dependent fashion. Combined use of anti-P-glycoprotein MAb with M-CSF or MCP-1 gene transduction caused complete inhibition of metastasis of H69/VP cells. The antimetastatic effect of these MAbs in vivo was mainly due to an antibody-dependent cell-mediated cytotoxicity reaction mediated by mouse macrophages. These findings suggest that the mouse-human chimeric MAb in combination with cytokine gene transduction may be useful for the eradication of lung cancer metastases in humans.
Masaki Hanibuchi, Seiji Yano, Yasuhiko Nishioka, Hiroaki Yanagawa, Tetsuya Kawano and Saburo Sone : Therapeutic efficacy of mouse-human chimeric anti-ganglioside GM2 monoclonal antibody against multiple organ micrometastases of human lung cancer in NK cell-depleted SCID mice, International Journal of Cancer, Vol.78, No.4, 480-485, 1998.
(要約)
The development of distant metastases to multiple organs is a critical problem in the treatment of human lung cancer. In this study, we evaluated the therapeutic efficacy of a mouse-human chimeric anti-ganglioside GM2 (GM2) monoclonal antibody (MAb), KM966 against metastasis formation of GM2-positive human lung cancer cells inoculated intravenously (i.v.) into natural killer (NK) cell-depleted severe combined immunodeficient (SCID) mice. GM2-positive human small cell lung cancer (SCLC), SBC-3 cells (1 x 10(6)), injected through a tail vein into NK cell-depleted SCID mice, formed large number of metastatic colonies in the liver, kidneys and lymph nodes by 42 days after inoculation (day 42). KM966, but not control MAb, given on days 2 and 7, almost completely inhibited metastasis formation of SBC-3 cells in the liver, kidneys and lymph nodes in a dose-dependent fashion. Moreover, treatment with KM966 at advanced stages of metastasis (even from day 28) significantly suppressed multiple organ metastases of SBC-3 cells. The anti-metastatic effect of KM966 in vivo was mainly due to an antibody-dependent cell-mediated cytotoxicity (ADCC) reaction mediated by macrophages of the SCID mice. Our findings suggest that the mouse-human chimeric anti-GM2 MAb, KM966 may be useful for eradicating multiple organ micrometastases of lung cancer in humans.
Punya Shrivastava, Masaki Hanibuchi, Seiji Yano, Prahlad Parajuli, Takashi Tsuruo and Saburo Sone : Circumvention of multidrug resistance by a quinoline derivative, MS-209, in multidrug-resistant human small-cell lung cancer cells and its synergistic interaction with cyclosporin A or verapamil, Cancer Chemotherapy and Pharmacology, Vol.42, No.6, 483-490, 1998.
(要約)
To develop a clinically useful approach to circumvent P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) in MDR human small-cell lung cancer (SCLC), we examined the ability of a novel quinoline compound, MS-209, to reverse MDR by inhibition of P-gp function in combination with other MDR-reversing drugs using a cytotoxicity assay. We established MDR human SCLC cells by culture in medium with gradually increasing concentrations of adriamycin (ADM). Compared with the parental human SCLC cells, SBC-3, the MDR variant SBC-3 cells obtained (SBC-3/ADM) were highly resistant to various chemotherapeutic agents due to P-gp expression. MS-209 reversed the resistance to ADM and vincristine (VCR) of SBC-3/ADM and H69/VP cells in a dose-dependent manner. Moreover, MS-209 in combination with cyclosporin A (CsA) or verapamil (VER) synergistically enhanced the antitumor effects of ADM and VCR on SBC-3/ADM cells. MS-209 restored ADM incorporation and this effect was enhanced by CsA and VER, suggesting that these synergistic effects were due to competitive inhibition of P-gp function. MS-209 in combination with CsA or VER might increase the efficacy of these chemotherapeutic agents against MDR human SCLC cells.
(キーワード)
Antineoplastic Agents / Carcinoma, Small Cell / Cyclosporine / Drug Resistance, Multiple / Drug Resistance, Neoplasm / Drug Synergism / Humans / Lung Neoplasms / P-Glycoprotein / Quinolines / Tumor Cells, Cultured / Verapamil
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 9788575
Yumi Ise, Hiroaki Yanagawa, Takanori Hirose, Kenji Tani, Seiji Yano, Yoshihiro Suzuki, Eiji Takeuchi, Kouji Maniwa, Eiji Shimizu, Fumitaka Ogushi, Toshiaki Sano and Saburo Sone : An autopsy case of cytokeratin 7-positive minute adenocarcinoma of the lung with systemic metastases, Internal Medicine, Vol.37, No.9, 766-769, 1998.
(要約)
We describe a 60-year-old woman with leg pain. Although metastatic bone tumor and atypical cells mimicking signet-ring cells in the bone marrow picture were observed, systemic survey revealed no primary lesion. The patient died two months after admission from systemic progress of the disease. Autopsy revealed a small focus of adenocarcinoma within the right upper lobe of the lung and systemic metastases without any particular changes in the gastrointestinal tract. The tumor cells of the lung were diffusely positive for cytokeratin 7, whereas cytokeratin 20 immunoreactivity was weak and focal, and that supported the lung origin of the present tumor. Moreover, the tumor cells in the bone marrow showed a similar pattern in immunoreactivity. These findings suggest that cytokeratin 7 and cytokeratin 20 immunoreactivity is helpful for the premortem diagnosis of the metastatic tumor of unknown origin.
(キーワード)
Biomarkers, Tumor / Bone Marrow / Bone Neoplasms / Carcinoma, Signet Ring Cell / Digestive System / Female / Fibula / Humans / Keratins / Lung / Lung Neoplasms / Middle Aged / Neoplasm Proteins / Neoplasms, Unknown Primary / Organ Specificity / Protein Isoforms / Radionuclide Imaging
Kenji Tani, Makoto Kashio, Nobuya Sano, Yoichi Nakamura, Fumitaka Ogushi and Saburo Sone : A case of sarcoidosis associated with chronic eosinophilic pneumonia, The Journal of Medical Investigation : JMI, Vol.45, No.1-4, 131-136, 1998.
(要約)
A 38-year-old man was hospitalized in our university hospital because of pulmonary opacities with bilateral hilar and mediastinal lymphadenopathy seen on chest radiograph. Eosinophilia was observed in the circulation and bronchoalveolar lavage (BAL) fluid. Histological examination revealed noncaseating epithelioid granulomas and eosinophilic infiltration in the lung. Based on these findings, a diagnosis of sarcoidosis combined with chronic eosinophilic pneumonia was made. The infiltrates on chest radiograph and BAL eosinophilia were promptly reduced with corticosteroid therapy, but only mild reduction was observed in diffuse nodular shadows and hilar and mediastinal lymphadenopathy, and high amounts of lymphocytes in BAL fluid remained. Increased IFN-gamma, IL-4 and IL-5 were detected in the BAL fluid, and corticosteroid therapy reduced IL-4 and IL-5 (Th-2 cytokines) but not IFN-gamma (Th-1 cytokine). These cytokine levels in BAL fluid were intimately correlated with the clinical course of sarcoidosis and chronic eosinophilic pneumonia.
Hiroaki Yanagawa, Kayo Shimada, Haruko Tanaka, Eiji Takeuchi, Kouji Maniwa, Yoshihiro Suzuki, Fumitaka Ogushi, Nobuya Sano, Keisuke Izumi and Saburo Sone : A case with Small Cell Carcinoma of the Esophagus:Measurement of Tumor Markers,Including Pro-gastrin-Releasing Peptide, Anticancer Research, Vol.18, No.4B, 2877-2880, 1998.
(要約)
Small cell carcinoma of the esophagus is a rare clinical entity and the accumulation of information is necessary to clarify its clinical behavior. We report a 69-year-old Japanese man with this rare disease with systemic metastases, including liver, bone and lymphnodes. The patient was treated with systemic chemotherapy consisting of 300 mg/m2 of carboplatin on day 1, and 80 mg/m2 of etoposide on days 1, 2 and 3. Although transient relief of subjective symptoms was obtained, the disease showed systemic progress, and the patient died on day 25 of chemotherapy. During the clinical course of the disease, serum pro-gastrin-releasing peptide (proGRP) decreased upon systemic chemotherapy from elevated level (54.6 pg/ml) to normal range (19.2 pg/ml). Further study is warranted to examine whether measurement of serum proGRP may yield valuable information on the diagnosis and monitoring activities of esophageal small cell carcinoma.
(キーワード)
Pro-gastrin-releasing peptide / small cell carcinoma / esophagus / chemotherapy / neuron specific enolase
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 9713479
Seiji Yano, Hiroaki Yanagawa, Masaki Hanibuchi, Kalpana Pai, Yasuhiko Nishioka, Takashi Tsuruo and Saburo Sone : The role of cyclosporin A on antibody-dependent monocyte-mediated cytotoxicity against human multidrug-resistant cancer cells, The Journal of Medical Investigation : JMI, Vol.44, No.3-4, 185-191, 1998.
(要約)
A P-glycoprotein (P-gp) inhibitor, cyclosporin A (CsA) was found to enhance the susceptibility of multidrug resistant (MDR) cancer cells to anti-P-gp antibody-dependent cellular cytolysis (ADCC) by monocytes, but the exact mechanism is unknown. In this study, we examined whether CsA enhanced the susceptibility of MDR cells through its inhibitory effect of P-gp function by using anti-ganglioside GM2 (GM2) monoclonal antibody (Ab), KM966, instead of anti-P-gp Ab, MRK16. Monocyte-ADCC induced by both KM966 and MRK16 against P-gp positive human MDR ovarian cancer cells was significantly augmented by addition of CsA. KM966, but not MRK16, induced monocyte-ADCC against P-gp negative human ovarian cancer cells and CsA enhanced this ADCC activity, indicating that suppressive effect of P-gp function by CsA was not essential to the enhancement of ADCC. Moreover, pretreatment of tumor cells with CsA augmented their susceptibility to monocyte-ADCC irrespective of P-gp expression. Interestingly, KM966 or MRK16 induced monocyte-ADCC against various human lung cancer cells expressing either GM2 or P-gp, but CsA did not affect these ADCC. These findings suggest that CsA may enhance the susceptibility to the monocyte-ADCC of ovarian cancer cells, but not of lung cancer cells, irrespective of its suppressive effect of P-gp function.
(キーワード)
Antibodies, Monoclonal / Cell Line / Cyclosporine / Cytotoxicity, Immunologic / Drug Resistance, Multiple / Humans / Monocytes / Neoplasms
Prahlad Parajuli, Seiji Yano, Masaki Hanibuchi, Hiroshi Nokihara, Tsutomu Shinohara and Saburo Sone : Effect of clarythromycin on the distant metastases of human lung cancer cells in SCID mice, The Journal of Medical Investigation : JMI, Vol.44, No.3-4, 205-210, 1998.
(要約)
Recently, the use of macrolides is suggested to be therapeutically effective in prolonging the survival of patients with inoperable non-small cell lung cancer. The purpose of this study was to examine therapeutic effects of a macrolide, clarythromycin (CAM) on the metastastic developments of two different human non-small cell lung cancers (squamous cell lung carcinoma RERF-LC-AI, and adenocarcinoma PC-14) in severe combined immunodeficient (SCID) mice depleted or undepleted of natural killer (NK) cells, respectively. CAM, injected subcutaneously at doses of 5 and 10 mg/kg body weight/day from day 7 to 41 after i.v. inoculation of human lung cancer cells, was not effective in inhibiting their distant organ metastases in SCID mice. CAM at concentrations of less than 10 micrograms/ml did not have a direct influence on the proliferation of these tumor cells in vitro. Although CAM alone was not effective in augmenting NK activity, it augmented the IL-2-induced killer (LAK) activity against Daudi cells in vitro. These results suggest that CAM alone may not be enough to control the spread of non-small cell lung cancer in the patient with T cell dysfunction.
Mineo Sone, Hiroyoshi Sei, Yusuke Morita, Takeshi Ogura and Saburo Sone : The Effects of Acetazolamide on Arterial Pressure Variability During REM Sleep in the Rat, Physiology & Behavior, Vol.63, No.2, 213-218, 1998.
(キーワード)
Acetazolamide / Arterial pressure / REM sleep / Rat
143.
Hiroaki Yanagawa, Takashi Haku, K Hiramatsu, Hiroshi Nokihara, Eiji Takeuchi, Seiji Yano, Masaki Hanibuchi and Saburo Sone : Intrapleural instillation of interferon gamma in patients with malignant pleurisy due to lung cancer, Cancer Immunology, Immunotherapy, Vol.45, No.2, 93-99, 1997.
(要約)
The effect of intrapleural instillation of recombinant human interferon gamma (IFN gamma) at increasing doses of (1-12) x 10(6) U was examined in six patients with cytologically positive pleural effusion due to lung cancer. Intrapleural instillation was repeated up to three times. Clinically, no reaccumulation of pleural effusion was observed in one patient and disappearance of lung cancer cells from the pleural effusion was seen in two other patients. No severe side-effects were observed. Considerable levels of IFN gamma remained in the pleural effusion as well as in patients' serum up to 7 days after instillation of 2 x 10(6) U and higher doses. The total cell number showed a transient decrease on day 1 of therapy. Levels of pro-inflammatory cytokines, such as tumor necrosis factor alpha, interleukin(IL)-1 beta and IL-6, in the pleural effusion remained almost stable after IFN gamma instillation. On the other hand, intrapleural IL-1 receptor antagonist levels were remarkably elevated by the instillation of IFN gamma. IL-2- and IL-12-inducible killer activity of pleural mononuclear cells tended to increase slightly. Despite the inability of IFN gamma to control pleural effusion in this treatment schedule, IFN gamma instilled by an intrapleural route had a potential local antitumor activity. Moreover, since IFN gamma persists in pleural effusions for a long time after a single instillation, such a therapy in combination with other fibrogenic biological response modifiers can be promising.
Yasuhiko Nishioka, Seiji Yano, Fujio Fujiki, Naofumi Mukaida, Kouji Matsushima, Takashi Tsuruo and Saburo Sone : Combined therapy of multidrug-resistant human lung cancer with anti-P-glycoprotein antibody and monocyte chemoattractant protein-1 gene transduction: The possibility of immunological overcoming of multidrug resistance, International Journal of Cancer, Vol.71, No.2, 170-177, 1997.
(要約)
We determined whether transduction of the monocyte chemoattractant protein-1 (MCP-1) gene into MDR human lung cancer cells affected their tumorigenicity and sensitivity to antibody-dependent cellular cytotoxicity (ADCC) reaction mediated by the anti-P-glycoprotein (P-gp) monoclonal antibody MRK16. The human MCP-1 gene inserted into an expression vector (BCMGSNeo) was transfected into MDR human small-cell lung cancer (H69/VP) cells. Monocyte chemotactic activity was found in culture supernatants collected from MCP-1-transfected H69/VP cells, but not in supernatants of parent and mock-transfected cells. In an in vitro experiment, recombinant MCP-1 did not affect monocyte-mediated ADCC against H69/VP cells when added to the monocyte culture in either the activation or the effector phase at sufficient concentrations to attract and activate monocytes. Tumorigenicity and growth rates of MCP-1-producing H69/VP cells in nude mice were similar to those of parental cells and mock-transfected cells. However, systemic treatment with MRK16 was more effective in inhibiting the formation of tumors by MCP-1-gene-transfected cells than by mock-transfected cells. Systemic treatment with MRK16 also inhibited the growth of a mixture (1:1) of MCP-1-producing cells and mock-transfected cells. These results suggest that combination therapy with MRK16 and MCP-1 gene transduction may be a useful immunological strategy to inhibit the growth of human MDR cancer cells expressing P-gp.
Seiji Yano, Yasuhiko Nishioka, Hiroshi Nokihara and Saburo Sone : Macrophage colony-stimulating factor gene transduction into human lung cancer cells differentially regulates metastasis formations in various organ microenvironments of natural killer cell-depleted SCID mice, Cancer Research, Vol.57, No.4, 784-790, 1997.
(要約)
We investigated whether local production of macrophage colony-stimulating factor (M-CSF), responsible for migration and activation of monocytes/macrophages at a tumor growth site, affected the metastatic pattern of lung cancer. For this, highly metastatic human squamous (RERF-LC-AI) or small (H69/VP) cell lung carcinoma cells were transduced with the human M-CSF gene inserted into pRc/CMV-MCSF to establish M-CSF-producing clones (MCSF-AI-9-18, MCSF-AI-9-24, and MCSF-VP-5). M-CSF gene transduction had no effect on the expression of surface antigen or on in vitro proliferation. After s.c. injection into SCID mice, the growth rates of M-CSF-producing cells were slower than those of parent or mock-transduced cells. In the metastatic model in SCID mice depleted of natural killer cells, RERF-LC-AI cells formed metastases mainly in the liver and kidneys, whereas H69/VP cells metastasized mainly to the liver and systemic lymph nodes. The numbers of metastatic colonies of MCSF-AI-9-18 and MCSF-AI-9-24 cells in the liver but not the kidneys were significantly reduced. The development of lymph node metastases of MCSF-VP-5 cells was also less than that of parent or mock-transduced cells. Treatment of SCID mice with anti-human M-CSF antibody resulted in a significant increase in liver metastases of their M-CSF gene transfectants. No significant differences were observed in the distributions in mice or in the in vitro invasive potentials of MCSF-AI-9-18 cells and Neo-AI-3 cells. These findings indicate that the antimetastatic effect of M-CSF may be specific to particular organs, suggesting the influence of heterogeneity of organ microenvironments on the metastasis of lung cancer.
Saburo Sone, Yoshihiko Yamamoto, Akio Adachi, Jin Asano, Shinya Iida and Yasuyoshi Saito : Functional Analysis of vif Genes Derived from Various Primate Immunodeficiency Viruses, Virus Genes, Vol.14, No.3, 195-200, 1997.
(要約)
Replication property in cells of human and simian immunodeficiency viruses (HIVs and SIVs) lacking intact vif gene was evaluated. Of 10 vif mutants constructed in vitro of the major four HIV/SIV groups, only those derived from HIV-1 and HIV-2/SIVmac displayed replication defect. The cell lines non-permissive for the vif mutants of HIV-1 and SIVmac were found to be different. To determine whether Vif is exchangeable between HIV-1 and SIVmac, chimeric virus clones with respect to the vi gene were constructed and virus replication in the cells non-permissive for the vif mutant viruses was monitored. Productive infection in these cells of chimeric viruses clearly indicated that Vif is functionally exchangeable, and that Vifs of different virus origin act through a similar mechanism.
Seiji Yano, Hiroshi Nokihara, Masaki Hanibuchi, Prahlad Parajuli, Tsutomu Shinohara, Tetsuya Kawano and Saburo Sone : Model of malignant pleural effusion of human lung adenocarcinoma in SCID mice, Oncology Research, Vol.9, No.11-12, 573-579, 1997.
(要約)
Malignant pleural effusion (PE) is a frequent problem in lung cancer. In this study, we established a model of malignant PE of human adenocarcinoma cells, PC-14, in SCID mice. Intravenously injected PC-14 cells formed colonies in the lungs as early as week 4 after tumor inoculation, and produced bloody PE in all recipient SCID mice by week 8. Pretreatment of SCID mice with anti-mouse IL-2 receptor beta chain antibody (TM-beta 1) to deplete natural killer (NK) cells markedly promoted the production of bloody PE and metastases to multiple organs, such as the lungs, liver, kidneys, and lymph nodes 4 weeks after tumor inoculation. Histological studies indicated that PC-14 cells formed colonies in the lungs, and then invaded the pleura and spread to the pleural cavity. To establish cell lines with a high potential to produce PE, we harvested PE, expanded the tumor cells in vitro, and injected them into SCID mice again. By four in vivo selection cycles in this way we obtained PC-14-PM4 cells, which produce lung metastases and PE earlier than PC-14 cells. The survival of SCID mice inoculated with PC-14-PM4 cells was significantly shorter than that of mice inoculated with PC-14 cells. The expressions of adhesion molecules, such as CD44, CD49d, ICAM-1, and MHC class I, on PC-14-PM4 cells tended to increase compared with PC-14 cells. These changes of adhesion molecules seem to be one of possible mechanisms involved in higher metastatic potential of PC-14-PM4 cells. PE models with PC-14 and PC-14-PM4 cells should be useful for biological and preclinical studies on malignant PE produced by human lung cancer.
Eiji Takeuchi, Hiroaki Yanagawa, Seiji Yano, Takashi Haku and Saburo Sone : Induction by interleukin-15 of human killer cell activity against lung cancer cell lines and its regulatory mechanisms, Japanese Journal of Cancer Research, Vol.87, No.12, 1251-1258, 1996.
(要約)
Interleukin (IL)-15 is a novel cytokine with IL-2-like activity. In the present study, we examined IL-15-mediated induction of killer activity of peripheral blood mononuclear cells (MNC) against lung cancer cell lines, and the regulatory mechanisms of this induction by IL-15. Cytotoxic activity was measured by 51Cr release assay. IL-15 at concentrations of more than 10 ng/ml induced significant killer activity of blood MNC against a small cell lung cancer cell line (SBC-3), as well as Daudi cells, and 50 ng/ml was considered its optimal concentration. A time course study revealed that an incubation period of 4-6 days was optimal for induction of killer activity. MNC cultured with IL-15 also exhibited killer activity against other lung cancer cell lines (H-69, N-291 and PC-9 cells). IL-15 and IL-12 had additive effects on induction of killer activity against SBC-3 cells. On the other hand, IL-15 had no synergistic or additive effect on induction of killer activity by IL-2. Fresh human monocytes isolated by centrifugal elutriation augmented the development of killer activity of lymphocytes stimulated by IL-15. As a humoral regulatory factor, IL-4 had a suppressive effect on induction of killer activity by IL-15. IFN-gamma, IL-1beta, TNF-alpha, IL-6 or IL-10 had no effect on induction of killer activity by IL-15 at the optimal concentration. These results suggest that IL-15 has potential for the immunotherapy of lung cancers.
Seiji Yano, Hiroaki Yanagawa, Yasuhiko Nishioka, Naofumi Mukaida, Kouji Matsushima and Saburo Sone : T helper 2 cytokines differently regulate monocyte chemoattractant protein-1 production by human peripheral blood monocytes and alveolar macrophages, The Journal of Immunology, Vol.157, No.6, 2660-2665, 1996.
(要約)
Th2 cytokines, such as IL-4, IL-10, and IL-13, suppress proinflammatory cytokine production by monocytes/macrophages. Since monocyte chemoattractant protein-1 (MCP-1) is presumed to play an important role in monocyte recruitment and activation during inflammatory and immune responses, we examined here the effects of these Th2 cytokines on MCP-1 production by human blood monocytes and alveolar macrophages. Unstimulated, highly purified blood monocytes did not produce MCP-1 spontaneously, while LPS treatment induced the production of MCP-1 and its mRNA expression. All Th2 cytokines tested suppressed LPS-induced MCP-1 production and its mRNA expression, although the suppressive effect of IL-13 was weaker than that of IL-4 or IL-10. In contrast, IL-10, but neither IL-4 nor IL-13, induced unstimulated peripheral blood monocytes to produce biologically active MCP-1 protein within 4 h, reaching a maximal level at 12 h. IL-10-induced MCP-1 production was reduced by pretreatment of IL-10 with anti-IL-10 Ab, negating the involvement of contaminated endotoxin. Moreover, IL-10 induced MCP-1 mRNA expression in unstimulated monocytes, independent of de novo protein synthesis. Furthermore, human alveolar macrophages produced MCP-1 spontaneously, and the production was inhibited by IL-4 or IL-13, but was augmented by IL-10. These findings suggest that IL-10 regulates MCP-1 production by monocytes/macrophages in a different way from other Th2 cytokines, such as IL-4 and IL-13, and contributes to host defense responses.
Hiroaki Yanagawa, Tetsuya Kawano, Takashi Haku, Seiji Yano, Kouji Maniwa and Saburo Sone : Palliative steriod therapy and serum interleukin-6 levels in a patient with lung cancer, Journal of Pain and Symptom Management, Vol.12, No.3, 195-198, 1996.
151.
Seiji Yano, Yasuhiko Nishioka, K Izumi, Takashi Tsuruo, T Tanaka, M Miyasaka and Saburo Sone : Novel metastasis model of human lung cancer in SCID mice depleted of NK cells, International Journal of Cancer, Vol.67, No.2, 211-217, 1996.
(要約)
Metastasis is a critical problem in the treatment of human lung cancer. Thus, a suitable animal model of metastasis of human lung cancer is required for in vivo biological and preclinical studies. In this study, we tried to establish a suitable model for this, using SCID mice. Neither human SCLC H69/VP cells (5 x 10(6)) nor squamous-cell carcinoma RERF-LC-AI cells (1 x 10(6)), injected through a tail vein, formed metastases in untreated SCID mice. Pre-treatment of SCID mice with anti-asialo GM1 serum resulted in only a few metastases of H69/VP cells, but pre-treatment with anti-mouse IL-2 receptor beta chain Ab (TM-beta 1) resulted in numerous lymph-node metastases 56 days after tumor inoculation. H69/VP-M cells, an in vivo-selected variant line, formed significant numbers of lymph-node metastases even in SCID mice pre-treated with anti-asialo GM1 serum. SCID mice depleted of NK cells by treatment with TM-beta 1 showed different patterns of metastasis when inoculated intravenously with the 2 different human lung cancer cell lines (H69/VP and RERF-LC-AI cells): H69/VP cells formed metastases mainly in systemic lymph nodes and the liver, whereas RERF-LC-AI cells formed metastases mainly in the liver and kidneys, with only a few in lymph nodes. A histopathological study showed that the metastatic colonies consisted of cancer cells. The numbers of metastatic colonies formed by the 2 cell lines increased with the number of cells inoculated. TM-beta 1 treatment of SCID mice efficiently removed NK cells from peripheral blood for at least 6 weeks, whereas, after treatment of the mice with anti-asialo GM1 serum, NK cells were recovered within 9 days. These findings suggest that NK-cell-depleted SCID mice may be useful as a model in biological and pre-clinical studies on metastasis of human lung cancer.
Saburo Sone, Takashi Tsuruo, S Sato, Seiji Yano, Yasuhiko Nishioka and Tsutomu Shinohara : Transduction of the macrophage colony-stimulating factor gene into human multidrug resistant cancer cells: enhanced therapeutic efficacy of monoclonal anti-P-glycoprotein antibody in nude mice, Japanese Journal of Cancer Research, Vol.87, No.7, 757-764, 1996.
(要約)
To develop a therapeutic modality for overcoming multidrug-resistant (MDR) cancer with anti-MDR1 antibody, we examined the effect of macrophage colony-stimulating factor (M-CSF) gene transfection into MDR AD10 cells on therapy of MDR cancer with anti-MDR1 antibody (MRK17) in nude mice. MDR human ovarian cancer (AD10) cells were transduced with the human M-CSF gene inserted into an expression vector to establish gene-modified cells capable of producing low (ML-AD10), intermediate (MM-AD10) nd high (MH-AD10) amounts of M-CSF. Systemic administration of MRK17 resulted in significant dose-dependent inhibition of subcutaneous growth of ML-AD10 tumors. In contrast, systemic administration of recombinant M-CSF in combination with MRK17 did not augment the therapeutic efficacy of MRK17 alone, but rather promoted the growth of the parent AD10 cells. To test the efficacy of in vivo M-CSF gene therapy combined with antibody, we mixed the parent AD10 cells with MH-AD10 cells producing a large amount of M-CSF, and inoculated the mixed cells subcutaneously. Treatment with MRK17 inhibited growth of the mixed cells more than that of the parent cells alone. Thus, combined therapy with anti-MDR1 mAb and M-CSF gene modification of MDR cancer cells may provide a new immunotherapeutic modality for overcoming MDR in humans.
Hiroaki Yanagawa, Seiji Yano, Takashi Haku, Y Ohmoto and Saburo Sone : Interleukin-1 receptor antagonist in pleural effusion due to inflammatory and malignant lung disease, The European Respiratory Journal, Vol.9, No.6, 1211-1216, 1996.
(要約)
Interleukin (IL)-1 is a key cytokine in inflammatory reactions. To clarify the mechanism of inflammation in the pleural cavity, we investigated the contribution of IL-1 and its antagonism to inflammatory processes in the pleural cavity. Interleukin-1 receptor antagonist (IL-1Ra) levels as well as IL-1 beta and interferon-gamma (IFN-gamma) levels were measured by enzyme immunoassay in pleural effusions from 70 patients. Pleural macrophages were also examined as possible sources of these cytokines in 10 patients. IL-1Ra was detectable in 28 patients (40%) out of 70 patients with pleural effusions. Patients with tuberculosis had significantly higher IL-1Ra as well as IFN-gamma levels in pleural effusion than patients with lung cancer. Transudative pleural effusions had low or undetectable IL-IRa levels. On the other hand, IL-1 beta levels were low, except in cases of parapneumonic pleural effusion. Spontaneous production of IL-1Ra pleural macrophages was observed in six patients, and IL-4 significantly augmented its production. Although spontaneous production of IL-1 beta was observed in only two patients, pleural macrophages produced significant amounts of IL-1 beta in response to lipopolysaccharide in all 10 patients examined. These results suggest that interleukin-1 receptor antagonist regulates various reactions by interleukin-1 in pleural effusion, and that pleural macrophages may act in situ as a source of interleukin-1 receptor antagonist.
Masaki Hanibuchi, Seiji Yano, Yasuhiko Nishioka, Hiroaki Yanagawa and Saburo Sone : Anti-ganglioside GM2 monoclonal antibody-dependent killing of human lung cancer cells by lymphocytes and monocytes, Japanese Journal of Cancer Research, Vol.87, No.5, 497-504, 1996.
(要約)
Ganglioside GM2 (GM2) frequently appears on the cell surface of human cancers of neuroendocrine origin. A mouse-human chimeric monoclonal antibody (mAb), KM966, against GM2 was previously found to promote the lysis of various cancer cells by human blood mononuclear cells (MNC). In this study, we analyzed the effector cells responsible for the chimeric mAb-dependent cell-mediated cytotoxicity (ADCC) against small cell lung cancer (SCLC) cells and examined the enhancing effect of various cytokines on the ADCC activity. The ADCC activity was assessed by 4-h 51Cr release assay. Highly purified lymphocytes (> 99%) and monocytes (> 90%) were separated by centrifugal elutriation from peripheral blood MNC of the same healthy donor. KM966 induced lysis of SCLC cells mediated by both lymphocytes and monocytes to similar extents, in a dose-dependent manner. Pretreatment of lymphocytes with various cytokines [interleukin (IL)-2, IL-12 and interferon-gamma] and that of monocytes with macrophage-colony-stimulating factor significantly augmented the killer activity against SCLC cells in the presence of KM966 mAb. KM966 was also effective for the lysis of non-small cell lung cancer cells in direct proportion to the GM2 expression levels. These findings suggest that combined treatment of KM966 mAb with cytokines may be therapeutically useful for in vivo killing of lung cancer cells expressing GM2 through the ADCC reaction.
Takashi Haku, Hiroaki Yanagawa, Y Ohmoto, Eiji Takeuchi, Seiji Yano, Masaki Hanibuchi, Hiroshi Nokihara, Naoki Nishimura and Saburo Sone : Systemic chemotherapy alters interleukin-1 beta and its receptor antagonist production by human alveolar macrophages in lung cancer patients, Oncology Research, Vol.8, No.12, 519-526, 1996.
(要約)
The purpose of this study was to determine whether cytotoxic chemotherapy influences the number and function of alveolar macrophages (AM) in patients with lung cancer. AM were obtained by bronchoalveolar lavage from 24 patients with lung cancer and 17 control patients. The functional integrity of AM was determined by their ability to produce interleukin-1 beta (IL-1 beta) and interleukin-1 receptor antagonist (IL-1ra) before and after platinum-containing systemic chemotherapy. The productions of IL-1 beta and IL-1ra were quantitated by enzyme immunoassays. The proportions of multinucleated cells among AM were significantly decreased after systemic chemotherapy in lung cancer patients. No significant difference in spontaneous and lipopolysaccharide (LPS)-stimulated IL-1 beta or IL-1ra production by AM was observed between lung cancer patients and control patients. Significant increase of IL-1 beta and significant decrease of IL-1ra production by AM were demonstrated in patients with small cell lung cancer who experienced response to systemic chemotherapy. These results suggest that systemic chemotherapy may influence functional roles of AM in the lung, and consideration of influence of systemic chemotherapy on host functions is important in cancer treatment.
Fumitaka Ogushi, Saburo Sone, Kenji Tani, Hiroko Takehara, Takeshi Endo, Takashi Haku, Yoichi Nakamura, Takeshi Ogura, Masaharu Kamada, Toshihiro Aono, Nobuya Sano and Yasukazu Ohmoto : Identification and localization of immunoglobulin binding factor in bronchoalveolar lavage fluid from healthy smokers, American Journal Respiratory and Critical Care Medicine, Vol.152, No.6, 2133-2137, 1995.
157.
Roustem Nabioullin, Hiroaki Yanagawa, Takashi Haku, K Hiramatsu, Seiji Yano, Masaki Hanibuchi, Kalpana Pai, Takashi Tsuruo and Saburo Sone : Influence of systemic chemotherapy on anti-P-glycoprotein antibody-dependent cell-mediated cytotoxicity in patients with small cell lung cancer, Japanese Journal of Clinical Oncology, Vol.25, No.4, 124-130, 1995.
(要約)
Anti-P-glycoprotein antibody (MRK-16)-dependent cell-mediated cytotoxicity (ADCC) by blood mononuclear cells (MNC) was examined in patients with small cell lung cancer (SCLC) before and after systemic chemotherapy. The effect of in vitro treatment of MNC with interleukin (IL)-2 and macrophage-colony-stimulating factor (M-CSF) was also examined. The ADCC reaction was assessed by a 6 h 51Cr-release assay using a multidrug-resistant (MDR) SCLC cell line (H69/VP cells). The MRK-16 monoclonal antibody was able to augment spontaneous cytotoxicity by MNC, even in SCLC patients. Pretreatment of MNC with IL-2 significantly augmented their ADCC ability in SCLC patients, while M-CSF had no effect on ADCC activity. After the first cycle of systemic chemotherapy, the ADCC activity tended to decline, but ADCC of MNC pretreated with IL-2 was not affected. The results suggest that anti-P-glycoprotein antibody, in combination with a cytokine such as IL-2, may be therapeutically useful against human SCLC resistant to chemotherapeutic drugs.
Seiji Yano, Saburo Sone, Yasuhiko Nishioka, Naofumi Mukaida, Kouji Matsushima and Takeshi Ogura : Differential effects of anti-inflammatory cytokines (IL-4, IL-10 and IL-13) on tumoricidal and chemotactic properties of human monocytes induced by monocyte chemotactic and activating factor., Journal of Leukocyte Biology, Vol.57, No.2, 303-309, 1995.
(要約)
The effect of recombinant human IL-4, IL-10, and IL-13 on the chemotaxis and antitumor activity of human blood monocytes induced by monocyte chemotactic and activating factor (MCAF) was examined. MCAF alone did not induce monocyte-mediated cytotoxicity against human melanoma (A375-M) cells whereas it significantly enhanced the cytotoxicity by norMDP-stimulated monocytes. MCAF, unlike IFN-gamma, had no priming effect on monocyte activation by norMDP. MCAF acted with norMDP or LPS to enhance the production of both IL-1 beta and TNF-alpha. Enhanced cytotoxicity of monocytes stimulated with MCAF plus norMDP was reduced by IL-1 receptor antagonist and anti-TNF-alpha antibody. IL-4, IL-10, and IL-13 suppressed the generation of antitumor activity and cytokine production (IL-1 beta and TNF-alpha) of monocytes stimulated with MCAF plus norMDP or LPS. Chemotaxis of monocytes induced by MCAF was not affected by norMDP or any of the anti-inflammatory cytokines (IL-4, IL-10, and IL-13). Moreover, the pretreatment of monocytes with anti-inflammatory cytokines did not suppress monocyte-chemotaxis. These findings suggest that in vivo recruitment and anti-tumor expression of blood monocytes induced by MCAF may be differently regulated by anti-inflammatory cytokines in vivo.
Hiroaki Yanagawa, Saburo Sone, Takashi Haku, Kazuto Mizuno, Seiji Yano, Y Ohmoto and Takeshi Ogura : Contrasting effect of interleukin-13 on interleukin-1 receptor antagonist and proinflammatory cytokine production by human alveolar macrophages, American Journal of Respiratory Cell and Molecular Biology, Vol.12, No.1, 71-76, 1995.
(要約)
We investigated the effect of interleukin-13 (IL-13) on production of IL-1 receptor antagonist (IL-1ra) and proinflammatory cytokines by human alveolar macrophages (AM). AM were obtained by bronchoalveolar lavage from healthy donors. The production of IL-1ra and proinflammatory cytokines, such as IL-1 beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha), were quantitated by enzyme immunoassays. AM spontaneously produced IL-1ra, and this production was significantly augmented by IL-13. On the other hand, IL-13 alone did not affect production of proinflammatory cytokines by freshly isolated AM. Upon stimulation with lipopolysaccharide (LPS), AM produced a significantly amount of proinflammatory cytokines as well as IL-1ra, but this production was suppressed by IL-13 in a dose-dependent manner. In contrast, IL-13 caused a small but reproducible increase in LPS-induced IL-1ra production. These regulatory effects of IL-13 were also observed in blood monocytes and macrophages generated in vitro by maturation of blood monocytes with granulocyte/macrophage colony-stimulating factor. These observations suggest that IL-13 may act as an anti-inflammatory cytokine through regulation of cytokine production by AM in the lung.
Hiroaki Yanagawa, Saburo Sone, Y Takahashi, Takashi Haku, Seiji Yano, Tsutomu Shinohara and Takeshi Ogura : Serum levels of interleukin 6 in patients with lung cancer, Journal of Japan Association of Breast Cancer Screening, Vol.71, 1095-1098, 1995.
161.
Roustem Nabioullin, Saburo Sone, Kazuto Mizuno, Seiji Yano, Yasuhiko Nishioka, Takashi Haku and Takeshi Ogura : Interleukin-10 is a potent inhibitor of tumor cytotoxicity by human monocytes and alveolar macrophages, Journal of Leukocyte Biology, Vol.55, No.4, 437-442, 1994.
(要約)
The effects of purified human interleukin-10 (IL-10) on the expression of antitumor activity of human monocytes and alveolar macrophages (AMs) obtained by centrifugal elutriation and bronchoalveolar lavage, respectively, from the same healthy donors were examined. Monocytes and AMs were incubated for 16 h in medium with lipopolysaccharide (LPS) in the presence or absence of IL-10 or IL-4, and then their tumoricidal activity was assayed by measuring 125I-IUdR release from human melanoma (A375) cells. Addition of IL-10 to cultures of monocytes or AMs with LPS resulted in dose-dependent suppression of their cytotoxicity against A375 cells, the suppression of the activity of monocytes being the higher. IL-10 also suppressed the synergistic effects of interferon-gamma and desmethyl muramyldipeptide in activation of monocytes. IL-10 inhibited the early induction phase of monocyte activation but not the effector phase (monocyte-mediated cytotoxicity). IL-10 plus IL-4 inhibited the antitumor activities of AMs and monocytes much more than either IL-10 or IL-4 alone. IL-10 and IL-4 at suboptimal concentrations also showed synergistic inhibitory effects. These findings suggest that IL-10 may be important in vivo in down-regulating the antitumor activities of monocytes and AMs in the lung by inhibiting their productions of antitumor effector molecules.
Saburo Sone, E Orino, Kazuto Mizuno, Seiji Yano, Yasuhiko Nishioka, Takashi Haku, A Nii and Takeshi Ogura : Production of IL-1 and its receptor antagonist is regulated differently by IFN-gamma and IL-4 in human monocytes and alveolar macrophages, The European Respiratory Journal, Vol.7, No.4, 657-663, 1994.
(要約)
Interleukin-4 (IL-4) has previously been found to downregulate interleukin-1 (IL-1) production, but to upregulate the production of IL-1 receptor antagonist (IL-1ra) in human monocytes stimulated with lipopolysaccharide (LPS). In the present study we wanted to determine whether the production of IL-1ra in human monocytes and alveolar macrophages (AMs) is regulated differently at the protein and messenger ribonucleic acid (mRNA) levels by IL-4 and interferon-gamma (IFN-gamma). AMs and monocytes obtained from healthy donors by bronchoalveolar lavage and centrifugal elutriation were stimulated with LPS in the presence or absence of IL-4 or IFN-gamma, and the expression of mRNA for IL-1 and IL-1ra was measured by Northern blot analysis. The production of IL-1 and IL-1ra was quantitated by enzyme immunoassays (EIAs). Spontaneous IL-1ra production was seen in AMs after incubation for 4 h in medium alone, but not in blood monocytes, at both the protein and mRNA levels. The spontaneous expression of the IL-1ra gene in AMs was augmented by incubation with IL-4. Interleukin-1 beta (IL-1 beta) production by LPS-stimulated AMs and monocytes was upregulated by IFN-gamma, but downregulated by IL-4. Interestingly, when stimulated with LPS, IFN-gamma inhibited IL-1ra production by monocytes, but up-regulated its production in human AMs at the protein and mRNA levels.(ABSTRACT TRUNCATED AT 250 WORDS)
Seiji Yano, Saburo Sone, Yasuhiko Nishioka, M Naito, Takashi Tsuruo and Takeshi Ogura : Cyclosporin A enhances susceptibility of multi-drug resistant human cancer cells to anti-P-glycoprotein antibody-dependent cytotoxicity of monocytes, but not of lymphocytes, Japanese Journal of Cancer Research, Vol.85, No.2, 194-203, 1994.
(要約)
Cyclosporin A (CsA) was previously found to bind to P-glycoprotein expressed on multidrug-resistant (MDR) cancer cells. In the present study, the effect of CsA on anti-P-glycoprotein monoclonal antibody (mAb)-dependent cell-mediated cytotoxicity (ADCC) against human MDR cells was examined. The ADCC reaction was assessed by 4-h 51Cr-release assay. Highly purified lymphocytes (> 99%) and monocytes (> 99%) obtained from blood mononuclear cells (MNC) of healthy donors were used as effector cells. CsA decreased the cytotoxic activity of MNC against MDR cells, but enhanced their ADCC activity in the presence of anti-P-glycoprotein mAb MRK16. Lymphocyte-mediated ADCC and natural killer activity against MDR cells were also suppressed by addition of CsA. CsA induced a significant dose-dependent increase in monocyte-mediated ADCC activity. Interestingly, pretreatment of MDR cancer cells, but not of monocytes, with CsA significantly enhanced ADCC activity mediated by monocytes, but not by lymphocytes. A CsA analog (PSC833) and FK-506, but not verapamil also increased the sensitivity of MDR cells to ADCC by monocytes. CsA did not affect the binding of monocytes to MDR cells in the presence of MRK16 mAb. These results indicate that CsA may directly enhance the susceptibility of MDR cancer cells to the monocyte-mediated ADCC reaction.
Yuji Heike, Saburo Sone, Seiji Yano, Hiroyuki Seimiya, Takashi Tsuruo and Takeshi Ogura : M-CSF gene transduction in multidrug-resistant human cancer cells to enhance anti-P-glycoprotein antibody-dependent macrophage-mediated cytotoxicity, International Journal of Cancer, Vol.54, No.5, 851-857, 1993.
(要約)
A human macrophage-colony-stimulating-factor (M-CSF) gene inserted into an expression vector (pRc/CMV-MCSF) was transfected into multidrug-resistant (MDR) human ovarian cancer cells (AD10) to induce secretion of human M-CSF into the medium. The M-CSF level in the culture medium of the transfected cells reached 100 ng/ml after 7 days, and the ability of the cells to secrete M-CSF was stable for at least 3 months. Transfection of the M-CSF gene did not result in any change in expression of MDRI (P-glycoprotein), proliferation or chemosensitivity of the cells from those of the parent cells. There was also no difference between the transfected and the parent cells in susceptibility to NK cell- or interleukin-2-activated killer-cell-mediated cytotoxicity. Human blood monocytes that had been incubated for 4 days in medium with the culture supernatant of MH-AD10 cells exhibited higher ADCC activity than untreated monocytes against MDRI-positive cancer cells. This effect of the supernatant of AD10 cells was completely abolished by its treatment with a monoclonal anti-M-CSF antibody (MAb). When transfected human MDR cells were injected into nude mice, an inverse correlation was seen between the ability of the cells to produce M-CSF and their tumorigenicity. Thus, gene modification of MDR cancer cells seems hopeful as a therapeutic method for enhancing anti-MDRI-MAb-dependent macrophage-mediated cytotoxicity against human MDR cancer cells.
K. Sugihara, Saburo Sone, Masayuki Shono, Akira Nii, K. Okumura and T. Ogura : Enhancement by monocytes of perforin production and its gene expression by human CD8+T cells stimulated with interleukin-2., Japanese Journal of Cancer Research, Vol.83, No.11, 1223-1230, 1992.
(キーワード)
T cells / monocytes
166.
N. Shimbara, E. Orino, Saburo Sone, t. Ogura, M. Takashina, Masayuki Shono, T. Tamura, H. Yasuda, K. Tanaka and Akira Ichihara : Regulation of gene expression of proteasomes (multi-protease complexes) during growth and differentiation of human hematopoietic cells., The Journal of Biological Chemistry, Vol.267, No.25, 18100-18109, 1992.
Yasuhiko Nishioka, Saburo Sone, E. Orino, A. Nii and T. Ogura : Down-regulation by interleukin 4 of activation of human alveolar macrophages to the tumoricidal state, Cancer Research, Vol.51, No.20, 5526-5531, 1991.
169.
Kenichi Maeda, Saburo Sone, Yasukazu moto Oh and Takeshi Ogura : A novel differentiation antigen on human monocytes that is specifically induced by granulocyte-macrophage colony-stimulating factor or IL-3, The Journal of Immunology, Vol.146, No.11, 3779-3784, 1991.
(要約)
A mouse mAb (TOMS-1) was generated against human blood monocytes that had been cultured for 4 days in medium with recombinant human granulocyte-macrophage CSF (GM-CSF). TOMS-1 (IgG1) detected a unique cell surface Ag with a molecular mass of about 43 kDa under both reducing and nonreducing conditions. TOMS-1Ag was expressed on monocytes treated with GM-CSF, but not on fresh or untreated monocytes. This Ag was induced dose dependently during culture of monocytes with GM-CSF for more than 24 h, reaching a maximum level in 3 or 4 days. Treatment of monocytes with cycloheximide in the presence of GM-CSF blocked TOMS-1Ag induction completely, indicating that de novo protein synthesis was required for its expression. TOMS-1Ag was also induced by treatment of monocytes with IL-3, but not with other cytokines such as macrophage-CSF, IL-4, and IFN-gamma or stimulators including LPS, desmethyl muramyl dipeptide, and PMA. TOMS-1Ag expression induced by GM-CSF was up-regulated by IL-4, but down-regulated by IFN-gamma. TOMS-1Ag was not induced on lymphocytes, granulocytes, or AM by GM-CSF or appropriate stimuli. TOMS-1Ag was also not expressed on any cell lines of human leukemias or solid tumors examined. Thus, TOMS-1Ag is a monocyte-specific differentiation Ag induced by GM-CSF or IL-3. These results suggest that TOMS-1 should be useful for monitoring the process of monocyte differentiation by GM-CSF or IL-3.
Saburo Sone, Eiji Kunishige, Farid Fawzy, Hiroaki Yanagawa, Akihiko Nii, Kenichi Maeda, Shinji Atagi, Yuji Heike, Yasuhiko Nishioka, Kazuhito Mizuno and Takeshi Ogura : Interleukin-2-inducible killer activity and its regulation by blood monocyte from autologous lymphocytes of lung cancer patigents, Japanese Journal of Cancer Research, Vol.82, No.6, 716-723, 1991.
(要約)
The ability of blood lymphocytes of newly diagnosed lung cancer patients to respond to interleukin 2 (IL-2) to become IL-2-activated killer (LAK) cells and its regulation by autologous monocytes were examined. LAK activity was measured by 51Cr release assay. The abilities of lymphocytes among blood mononuclear cells (MNC) of subjects of different ages without malignancies to generate LAK activity against NK-cell resistant Daudi cells and lung adenocarcinoma (PC-9) cells were very similar. The LAK activity of blood MNC of lung cancer patients was also nearly the same as that of blood MNC of control subjects. There was no significant difference in IL-2-inducible LAK activity between MNC of patients with small cell lung cancer (SCLC) and those of patients with non-SCLC. Monocytes and lymphocytes were separated from blood MNC on a one-step Percoll gradient. Monocytes of lung cancer patients were found to augment in vitro induction of LAK activity by IL-2 of autologous blood lymphocytes. In contrast, endotoxin-stimulated monocytes suppressed LAK induction of autologous lymphocytes of cancer patients. These findings suggest that administration of IL-2 and LAK cells induced in vitro may be of benefit in the treatment of lung cancer.
Chikamasa Yamashita, Saburo Sone, Takeshi Ogura and Hiroshi Kiwada : Potential Value of Cetylmannoside-modified Liposomes as Carriers of Macrophage Activators to Human Blood Monocytes, Japanese Journal of Cancer Research, Vol.82, No.5, 569-576, 1991.
(要約)
The present study was undertaken to examine the potential value of cetylmannoside-modified multilamellar liposomes (Man-MLV) as carriers for transfer of macrophage activators to blood monocytes. Highly purified blood monocytes were isolated by centrifugal elutriation from healthy donors under endotoxin-free conditions. Freshly prepared monocytes phagocytosed Man-MLV to a lesser extent than monocyte-derived macrophages, but they took up Man-MLV much more effectively than control liposomes without cetylmannoside (control MLV). Phagocytosis of Man-MLV, but not control MLV by monocytes was inhibited by addition of D-mannose, but not of D-galactose. Desmethyl-muramyl dipeptide (norMDP) entrapped in Man-MLV was far more effective than norMDP entrapped in MLV in activating monocytes to the tumoricidal state. The effect of encapsulation of recombinant human macrophage colony-stimulating factor (M-CSF) in Man-MLV on prolongation of survival of monocytes was examined. Blood monocytes that had been incubated for up to 21 days with Man-MLV containing 5-20 U of M-CSF per ml were effective in prolonging monocyte survival, but monocytes that had been incubated in medium with less than 50 U/ml of M-CSF or with control MLV containing 5-10 U of M-CSF showed no increase of monocyte survival over that in medium alone. Addition of rabbit anti-M-CSF antiserum did not affect survival prolongation of monocytes by M-CSF encapsulated in Man-MLV. We conclude that liposomes modified with cetylmannoside are far more effective than unmodified liposomes as a carrier to deliver biological response modifiers to human blood monocytes.
Hiroaki Yanagawa, Saburo Sone, Akihiko Nii, Katsuyuki Fukuta, Mie Nakanishi, Kenichi Maeda, Mitsuo Honda and Takeshi Ogura : Lymphokine-activated killer induction and its regulation by macrophages in malignant pleural effuusion, Japanese Journal of Cancer Research, Vol.80, No.12, 1220-1227, 1989.
(要約)
Mononuclear cells (MNC) from pleural effusions and peripheral blood of 18 patients with primary lung cancer with malignant pleural effusion were studied. Pleural and blood MNC generated lymphokine-activated killer (LAK) activity similarly when cultured for 4 days with an optimal concentration of interleukin 2 (IL-2). Highly purified lymphocytes (greater than 98%) and monocyte-macrophages (greater than 90%) were isolated by discontinuous Percoll gradient centrifugation from pleural and blood MNC. Pleural macrophages, as well as blood monocytes, showed significant augmenting effects on in vitro LAK cell induction from pleural and blood lymphocytes by IL-2. During daily intrapleural administration of IL-2, significant induction of LAK activity in vivo was observed after 3 days, but then this LAK activity in pleural MNC decreased almost to zero by day 15. Daily injections of IL-2 resulted in reduction in the up-regulation of LAK induction by pleural macrophages and also in increases in the levels of soluble IL-2 receptors in pleural effusions. These findings indicate that in vivo LAK induction of lymphocytes in malignant effusions by IL-2 may be regulated by macrophages in the effusions.
Itsuo Katoh, Yoshiyasu Egawa, Tetsuya Kitagawa, Hiroki Taki, Masanori Yoshizumi, Saburo Sone, Hiroaki Yanagawa, T Ogura, A Satoh and Y Natori : Intraarterial lymphocyte injection therapy for lyphedema of the extremities, Progress in Lymphology-, Vol.12, 81-84, 1989.
179.
Kenji Tani, Fumitaka Ogushi, Saburo Sone, Noriko Kagawa and Takeshi Ogura : Chylothorax and chylous ascites in a patient with uterine cancer, Japanese Journal of Clinical Oncology, Vol.18, No.2, 175-182, 1988.
Saburo Sone, K. Tachibana and Masayuki Shono : Potential value of liposomes containing muramyl dipeptide for augmenting the tumoricidal activity of human alvolar macrophages, Joranal of Biologcal Response, Vol.3, No.2, 185-194, 1984.
The purpose of this article was to understand the psychological conditionof the adult patient had received the steroid therapy for some diseases inhospital.A psychological factor of six patients was investigated by "AnxietyCognition related to Sickness Questionnaire" and "General HealthQuestionnaire GHQ-12"in study 1.In study 2, three patients who had highanxiety cognition and low mental health degree was interviewed byNarrative Approach(NA).As the results of these studies, we presumed thatthe patients needed some supports according to them,because the patientshad high anxiety and stress in hospital.And,it was suggested that the NAinterview was effective method for assess patient's psychological side.Andthe rewriting the patient's story by NA interview could improve patient'smental health.
(キーワード)
Narrative Approach / steroid therapy / anxiety for physical disease
Mari Miki, Youichi Nakamura, Saburo Sone, Mitsugu Ikebe, Sumiko Yoshinaga, Eriko Harada, Fumiko Michishige, Toshiko Terao, Hiroshi Maezawa and Susumu Yasuoka : Effect of human airway trypsin-like protease (HAT) on proliferation of normal bronchial epithelial cells and normal lung fibroblasts., Proceeding of Airway Secretion research, Vol.3, 15-24, 2001.
Toshifumi Tezuka, Masahiko Azuma, Hisatugu Goto, Yasuhiko Nishioka and Saburo Sone : Psychiatric symptoms in a patient with Churg-Strauss syndrome., Internal Medicine, Vol.51, No.3, 301-303, Feb. 2012.
(要約)
We report a case of Churg-Strauss syndrome (CSS) in a patient with multiple cerebral infarctions and psychotic symptoms. A 67-year-old man presented a high-grade fever and delirium. He was clinically diagnosed with Churg-Strauss syndrome on the basis of the presence of asthma, neuropathy, blood eosinophilia, and increased myeloperoxidase-specific anti-neutrophil cytoplasmic antibody (MPO-ANCA) activities. Though multiple cerebral infarctions are irreversible, this patient's psychiatric symptoms improved with steroid treatment. Psychiatric symptoms associated with CSS are very rare.
Seiji Yano, W Wang, Q Li, T Yamada, S Takeuchi, K Matsumoto, Yasuhiko Nishioka and Saburo Sone : SHGF-MET in resistance to EGFR tyrosine kinase inhibitors in lung cancer., Current Signal Transduction Therapy, Vol.6, No.2, 228-233, May 2011.
Recent insights into the molecular mechanism of cancer progression have given rise to specific target-directed therapies, including monoclonal antibodies and small molecular compounds, and the advent of target-specific therapeutics has remarkably improved the outcomes of patients with various malignancies. Recent advance also lead to the identification of prognostic biomarkers as predictive factors in determining response to molecular targeted drugs. Future studies also need to develop biomarkers to further increase the power of patient selection for molecular targeted therapy. Here we review the recent progress in developing new molecular targeted drugs and the resistance to treatments as well as the importance of measuring the QOL by patient-reported outcome, for personalized therapy.
The 12 th Oncology Forum discussed the progress and future strategy of cancer prevention in Japan. The National Cancer Center has established a research center for screening focusing on the most common six cancer, stomach, lung, liver, colon, breast and uterus cancer. The program so far had a cumulative detection rate of 3.3%, which is high,and may reflect the selection of subjects. Screening and chemoprevention is also being investigated in prostate cancer, but the issues centre on how to make this widely available. High risk subjects can also be identified for breast cancer. Obesity and family history are especially important. In colorectal cancer studies are evaluating different diets, but general application is not yet possible and the infrastructure to implement any general screening and prevention does not exist. Development of pharmaceutical treatments for prevention is difficult because of the need for very safe treatments, and also because of the length of time needed to carry out studies. Overall, cancer prevention is still in evolution. New approaches are needed, and new infrastructure will be needed at a government level to implement this.
(キーワード)
Breast Neoplasms / Chemoprevention / Clinical Trials as Topic / Colorectal Neoplasms / Drug Industry / Female / Humans / Male / Medical Oncology / Neoplasms / Prostatic Neoplasms / Uterine Neoplasms
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16227757
Kenji Tani, Keiko Sato and Saburo Sone : The Role of Chemokines in the Pathogenesis of Rheumatoid Arthritis, Current Rheumatology Reviews, Vol.1, No.2, 143-150, Jun. 2005.
The purpose of cancer screening is to widen the difference between morbidity and mortality of the target cancer. Since 1983 cancer screening has been supported by the Japanese government. Initially it covered gastric and cervical cancer with lung, breast and endometrial cancers supported from 1987 and colorectal cancer in 1992. Since 1998 support for cancer screening has been transferred to local government. It is generally accepted that the uptake of screening services is too low. Estimates for Japan suggest that at best 30% of the eligible population accept the services. In other parts of the world screening is more widely accepted, for example, 67% for breast cancer and 79% for cervical screening in the USA. Barriers within Japan for increasing screening are complex and include, legal, ethical, financial, technical infrastructure, data related matters and the level of understanding and education of the general public. In 2000 the MHLW conducted an evaluation of cancer screening in terms of usefulness and effectiveness. It concluded that fair or better evidence for reduction in mortality existed for cervical cancer (cytology), breast cancer (mammography with clinical examination for women aged > or = 50 years), colorectal cancer (faecal occult blood test), gastric cancer, lung cancer and liver cancer. As a step towards improving screening services and their uptake by the general public a new Research Centre for Cancer Prevention and Screening was established at the National Cancer Centre in October 2003. This and other initiatives will build on the progress of the past 20 years but it is generally agreed that there is still have a long way to go.
Based on presentations on the basic concepts and scientific rationale of anti-angiogenic approaches to cancer therapy and the possible applications in the area of prostate cancer, gastrointestinal cancer, lung cancer and breast cancer it is easy to conclude that development of anti-angiogenic approaches into clinical therapies is extremely challenging. It is now well established that cancer growth is increased by angiogenic factors and that inhibition of angiogenesis decreases growth and metastatic potential. Anti-angiogenic effect can be obtained through interference with multiple targets. Further development of new strategies involving such novel cancer therapies requires wide reaching development of translational research abilities. However, for moving new therapies into the clinic same rigorous criteria need to be applied as is done for traditional therapies. Angiogenesis appear to be a critical factor for development of prostate, gastric, lung and breast cancers. Development of new anti-angiogenic treatment modalities might become very important in these diseases. A critical requirement for the successful clinical development will be the development of imaging techniques that can help evaluate the effect on blood vessel functionality. Such surrogate markers of anti-angiogenic effect will be essential for optimising molecules and doses.
Hirokazu Ogino, Hisatsugu Goto, Okano Yoshio, Machida Hisanori, Hatakeyama Nobuo, Ogushi Fumitaka, Haku Takashi, Takanori Kanematsu, Tomoyuki Urata, Souji Kakiuchi, Masaki Hanibuchi, Saburo Sone and Yasuhiko Nishioka : Phase II study of S-1 with patient-reported outcome evaluation in elderly patients with previously untreated advanced non-small cell lung cancer., ERS International Congress 2017, Milan, Italy, Sep. 2017.
2.
Hisatsugu Goto, Ogushi Fumitaka, Haku Takashi, Tomoyuki Urata, Takanori Kanematsu, Souji Kakiuchi, Masaki Hanibuchi, Saburo Sone and Yasuhiko Nishioka : Phase study of S-1 with patient-reported outcome evaluation in elderly patients with advanced non-small cell lung cancer., 18th Congress of the Asian Pacific Society of Respirology, Yokohama, Nov. 2013.
3.
Mitsuhashi Atsushi, Hisatsugu Goto, Kuramoto Takuya, Tabata Sho, Yukishige Sawaka, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, Ledford G Julie, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in nude mice via modulating host immune response., 18th Congress of the Asian Pacific Society of Respirology, Yokohama, Nov. 2013.
4.
Hisatsugu Goto, Mitsuhashi Atsushi, Kuramoto Takuya, Tabata Sho, Yukishige Sawaka, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, Ledford G. Julie, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses lung cancer progression by regulating tumor-associated macrophage polarization., ATS 2013 Conference, Philadelphia, May 2013.
5.
Atsushi Mitsuhashi, Hisatsugu Goto, Trung The Van, Takuya Kuramoto, Sho Tabata, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, Saburo Sone and Yasuhiko Nishioka : Acquired resistance to anti-VEGF therapy via angiogenic switch in orthotopically implanted human malignant pleural mesothelioma in SCID mice., Ninth AACR-Japanese Cancer Association Joint Conference: Breakthroughs in Basic and Translational Cancer Research, Maui, Feb. 2013.
6.
Sawaka Yukishige, Hisatsugu Goto, Atsushi Mitsuhashi, Takuya Kuramoto, Sho Tabata, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, Julie G. Ledford, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses lung cancer progression by regulating tumor-associated macrophage polarization., Ninth AACR-Japanese Cancer Association Joint Conference: Breakthroughs in Basic and Translational Cancer Research, Maui, Feb. 2013.
7.
Atsushi Mitsuhashi, Hisatsugu Goto, Takuya Kuramoto, Sho Tabata, Sawaka Yukishige, Shinji Abe, Masaki Hanibuchi, Souji Kakiuchi, Atsuro Saijo, Yoshinori Aono, Hisanori Uehara, Seiji Yano, Julie G Ledford, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in nude mice via modulating host immune response, Joint Meeting to Seoul National University, Seoul, Dec. 2012.
8.
Takuya Kuramoto, Hisatsugu Goto, Atsushi Mitsuhashi, Sho Tabata, Hirohisa Ogawa, Hisanori Uehara, Atsuro Saijo, Souji Kakiuchi, Masaki Hanibuchi, Yoichi Maekawa, Koji Yasutomo, Shin-ichi Akiyama, Saburo Sone and Yasuhiko Nishioka : Dll4-Fc, an inhibitor of Dll4-notch signaling, suppresses liver metastasis of small cell lung cancer cells through down-regulation of NF-kappa-B activity, 14th International Biennial Congress of the Metastasis Research Society, Brisbane, Sep. 2012.
9.
Hisatsugu Goto, Atsushi Mitsuhashi, Takuya Kuramoto, Masaki Hanibuchi, Souji Kakiuchi, Sho Tabata, Atsuro Saijo, Jo R Wright, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in an experimental lung metastasis model, 14th International Biennial Congress of the Metastasis Research Society, Brisbane, Sep. 2012.
10.
Hisatsugu Goto, Atsushi Mitsuhashi, Takuya Kuramoto, Masaki Hanibuchi, Souji Kakiuchi, Sho Tabata, Atsuro Saijo, Shin-ichi Akiyama, Jo R Wright, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in an experimental lung metastasis model, ATS 2012 International Conference, San Francisco, May 2012.
11.
Yasuhiko Nishioka, Shinji Abe, Mika K Kaneko, Yoshinori Aono, Jun Huang, Hisatsugu Goto, Masatoshi Kishuku, Masaki Hanibuchi, Saburo Sone, Kazuo Minakuchi and Yukinari Kato : Antitumor effects of anti-podoplanin antibody NZ-1 against malignant mesothelioma via ADCC, ATS 2012 International Conference, San Francisco, May 2012.
12.
Takuya Kuramoto, Hisatsugu Goto, Atsushi Mitsuhashi, Sho Tabata, Hirohisa Ogawa, Hisanori Uehara, Seidai Sato, Souji Kakiuchi, Masaki Hanibuchi, Yoichi Maekawa, Koji Yasutomo, Shin-ichi Akiyama, Saburo Sone and Yasuhiko Nishioka : Dll4-Fc, an inhibitor of Dll4-notch signaling, suppresses liver metastasis of small cell lung cancer cells through down-regulation of NF-kappa-B activity, AACR Annual Meeting 2012, Chicago, Apr. 2012.
13.
Katsuhiro Kinoshita, Yasuhiko Nishioka, Masami Kishi, Momoyo Azuma, Yoshinori Aono, Hisanori Uehara, Akio Takezaki, S. Hatakeyama, E. Kawahara and Saburo Sone : Antifibrotic effects of focal adhesion kinase inhibitor in bleomycin-induced pulmonary fibrosis in mice., ATS 2011 International Conference, Denver, May 2011.
14.
Yoshinori Aono, J.G. Ledford, Yasuhiko Nishioka, Saburo Sone, M.F. Beers and J.R. Wright : SP-D attenuates bleomycin induced lung fibrosis via regulating recruitment and activation of macrophages and fibrocytes., ATS 2011 International Conference, Denver, May 2011.
15.
Atsushi Mitsuhasi, Hisatsugu Goto, Takuya Kuramoto, Souji Kakiuchi, Seidai Sato, Van The Trung, Atsuro Saijo, Makoto Tobiume, Kenji Otsuka, Yasuhiko Nishioka, Rae Jo Wright and Saburo Sone : Surfactant protein A suppresses progression of human lung adenocarcinoma in an experimental lung metastasis model, ATS 2011 International Conference, Denver, May 2011.
16.
Momoyo Azuma, Yasuhiko Nishioka, El-morshedy Mohammed Reham, Jun Kishi, Masami Kishi, Katsuhiro Kinoshita, Hisanori Uehara, Keisuke Izumi and Saburo Sone : The role of sphingosine 1-phospate (SIP) signaling in pulmonary fibrosis., ATS 2011 International Conference, Denver, May 2011.
17.
Yasuhiko Nishioka, Hisatsugu Goto, Souji Kakiuchi, Masaki Hanibuchi, Yano Seiji and Saburo Sone : Bone metastasis of lung cancer and its molecular-targeted therapy., Joint Metastasis Research Society-AACR Conference, Philadelphia, Sep. 2010.
18.
Huang Jun, Yasuhiko Nishioka, Shinji Abe, Morita Yuki, Ozaki Shuji, Verma Kumar Vinod, Kishuku Masatoshi, Minakuchi Kazuo, Sekido Yoshitaka, Matsumoto Toshio and Saburo Sone : Anti-HM1.24 antibodies induce antibody-dependent cellular cytitixicity against mesothelioma cells., IMIG2010 (The 10th International Conference of the International Mesothelioma Interest Group), Kyoto, Sep. 2010.
19.
Trung The Van, Masaki Hanibuchi, Hisatsugu Goto, Souji Kakiuchi, Kuramoto Takuya, Sato Seidai, Tobiume Makoto, Atsuro Saijo, Tabata Sho, Yasuhiko Nishioka, Shin-ichi Akiyama and Saburo Sone : TS-1 suppresses the growth of malignant pleural mesithelioma cells co-expressing dihydropyrimidine dehydrogenase and thymidine phosphorylase in an orthotopic model., The 10th International Conference of the International Mesothelioma Interest Group, Sep. 2010.
20.
Khalil Hamed Abdellah, Hiromitsu Takizawa, Kazuya Kondo, Matsuoka Hisashi, Hiroaki Toba, Yasushi Nakagawa, Koichiro Kenzaki, Shoji Sakiyama, Kakiuchi Soji, Saburo Sone and Akira Tangoku : In vitro and in vivo photodynamic diagnosis using 5-aminolevulinic acid in malignant mesothelioma, The 10th International Conference of the International Mesothelioma Interest Group, Aug. 2010.
21.
Momoyo Azuma, Takada Shinsuke, Sato Seidai, Hisatsugu Goto, Hiroaki Yanagawa, Yasuhiko Nishioka, Kenichi Maeda, Shin-ichi Akiyama and Saburo Sone : Investigation of complementary and alternative medicine in lung cancer patients in Tokushima Univerity Hospital., 第9回アジア臨床腫瘍学会学術集会, Gifu, Aug. 2010.
22.
Kishi Masami, Yasuhiko Nishioka, Momoyo Azuma, Yoshinori Aono, Batmunkh Rentsenkhand, El-morshedy Mohammed Reham, Katsuhiro Kinoshita, Jun Kishi, Uehara Hisanori, Izumi Keisuke and Saburo Sone : Anifibrotic effects of APA5 and APB5, blocking antibodies specific for PDGF receptors on bleomycine-induced pulmonary fibrosis in mice., ATS 2010 International Conference, New Orleans, May 2010.
23.
Momoyo Azuma, Yasuhiko Nishioka, El-morshedy Mohammed Reham, Jun Kishi, Katsuhiro Kinoshita, Uehara Hisanori, Izumi Keisuke and Saburo Sone : Effects of the novel immunomodulatory sphingosine l-phospate(S1P) receptor agonist FTY720 in bleomycine-induced pulmonaty fibrosis in mice., ATS 2010 International Conference, New Orleans, May 2010.
24.
Saburo Sone, Soji Kakiuchi and Seiji Yano : Real-time PCR-based biomarker to predict the sensitivity of non-small cell lung cancer(NSCLC) to EGFR-TKI, The 10th International Symposium on Cancer Chemotherapy, Tokyo, Jan. 2006.
25.
Junya Miyata, Kenji Tani, Keiko Sato, Shinsaku Otsuka, Tomoyuki Urata, Shizuka Shigekiyo, Battur Lkhagvas, Nobuya Sano and Saburo Sone : The significance in rheumatoid arthritis as a monocyte chemoattractant, 2005 American College of Rheumatology(ACR/ARHP) Annual Scientific Meeting, San Diego, Nov. 2005.
26.
M. Azuma, Yasuhiko Nishioka, Yoshinori Aono, M. Inayama, H. Makino, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Inhibition of bleomycin-induced pulmonary fibrosis in mice by imatinib (Gleevec) and erythromycin-a role of aipha-1-acid glycoprotein, 100th American thoracic society (ATS), San Diego, May 2005.
27.
Yasuhiko Nishioka, M. Inayama, H. Makino, M. Ugai, Yoshinori Aono, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Anti-fibrotic effect of IKKB inhibitor IMD-0354 in bleomycin-induced pulmonary fibrosis, 100th American thoracic society (ATS), San Diego, May 2005.
28.
Mami Inayama, Yasuhiko Nishioka, Masahiko Azuma, Yoshinori Aono, Hideki Makino, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Anti-fibrotic effect of IKKB inhibitor IMD-0354 in bleomycin-induced pulmonary fibrosis, 100th American Thoracic Society Annual Meeting, San Diego, May 2005.
29.
Seiji Yano, Emiko Nakataki, Hiroaki Muguruma, Hideki Tomimoto, Soji Kakiuchi and Saburo Sone : Establishment of orthotopically implanted malignant mesothelioma model which represents intrathoracic tumors and bloody pleural effusions, 96th Annual Meeting of American Association for Cancer Research, Anaheim, Apr. 2005.
30.
Hiroaki Muguruma, Seiji Yano, Emiko Nakataki, Hideki Tomimoto, Takafumi Todoroki, Soji Kakiuchi, Makoto Kawatani, Hiroyuki Osada and Saburo Sone : Bone metastasis inhibition by RMA, 96th Annual Meeting of American Association for Cancer Research, Anaheim, Apr. 2005.
31.
Saburo Sone : Genetic Predictors of EGF Receptor-targeted therapy as personalized medicine for Non-small Cell lung cancer patients, the 12th Catholic Respiratory and Allergy Workshop, Seoul, Oct. 2004.
32.
Saburo Sone : Microarrays to predict organotropism of metastasis, the 10th International Congress of Metastasis Research Society(9/17-20), Genova, Sep. 2004.
33.
Soji Kakiuchi, Seiji Yano, Toyokazu Miki, Yusuke Nakamura and Saburo Sone : Genome-wide screening of the genes correlates with the cross-talk between metastatic cancer cells and microenvironment in small cell lung cancer cells, the 10th International Congress of Metastasis Research Society, Genova, Sep. 2004.
34.
Seiji Yano, Rui Zheng, Soji Kakiuchi, Yuka Matsumori, Emiko Nakataki, Hiroaki Muguruma and Saburo Sone : Sre tyrosine kinase inhibitor,M475271,suppresses subcutaneous growth and production of lung metastasis by inhibition of proliferation,invasion,and vascularization of human lung adenocarcinoma cells, the 10th International Congress of Metastasis Research Society(9/17-20), Genova, Sep. 2004.
35.
Saburo Sone : Bone metastasis of lung cancer and its molecular-based therapy, The 3rd Sino-Japan Joint Conference for Cancer Reasearch(9/3-4), Xi'an, Sep. 2004.
36.
Yoshinori Aono, Yasuhiko Nishioka, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Anti-fibrotic effect of STI571(Gleevec) in bleomycin-induced pulmonary fibrosis, 99th American thoracic society (ATS), Orlando, May 2004.
37.
Yasuhiko Nishioka, K. Manabe, J. Kishi, M. Inayama, Yoshinori Aono, M. Ugai, Kenji Tani and Saburo Sone : Analysi of CXCR3 ligands, IP-10, MIG and I-TAC, in patients with pulmonary sarcoidosis, 99th American thoracic society (ATS), Orlando, May 2004.
38.
Yasuhiko Nishioka, Yoshinori Aono, M. Inayama, M. Ugai, J. Kishi, Hisanori Uehara, Keisuke Izumi and Saburo Sone : Imatinib as a novel PDGF receptor-targeted drug to prevent bleomycin-induced lung fibrosis, 第44回日本呼吸器学会総会 International progam 3, Tokyo, Apr. 2004.
39.
Saburo Sone : Genetic Predictors of Egf Receptor Tyrosine kinase-targeted Therapy for Non-small Cell lung cancer, 6th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association Advances in Cance Resarch, Hawaii, Jan. 2004.
40.
Saburo Sone, Soji Kakiuchi, Seiji Yano, Yataro Daigo, Masahiro Fukuoka and Yusuke Nakamura : Genetic predictors of EGF receptor tyrosine kinase-targeted therapeutics for Non-small lung cancer, The 8th International Symposium on Cancer Chemotherapy(12/1-2), Tokyo, Dec. 2003.
41.
Saburo Sone, Soji Kakiuchi, Seiji Yano, Takashi Tsuruo and Yosuke Nakamura : Genetic predictor of molecular-targeted therapy for non-small cell lung cancer, 3ed Japan-Usa Cancer Treatment Symposium, Kyoto, Oct. 2003.
42.
Yuka Matsumori, Hisatsugu Goto, Emiko Nakataki, Takanori Kanematsu, Seiji Yano, S Wedge, A Ryan and Saburo Sone : ZD6474,an inhibitor of vascular endothelial growth factor receptor tyrosine kinase activity,Inhibits growth of human non-small cell lung cancer metastasis that are resistant to treatment with gefitinib("Iressa",ZD1839), 94th Annual Meeting of American Association for Cancer Reserch, Washington, D.C., Jul. 2003.
43.
Yasuhiko Nishioka, Jun Kishi, K. Manabe, M. Miki, Y. Makamura, F. Ogushi, T. Sano, H. Bando, Kenji Tani and Saburo Sone : Analyses of Th1-and Th2-Type Chemokines in Bronchoalveolar Lavage Fluids from Patients with Interstitial Lung Diseases, 99th American thoracic society (ATS), Seattle, May 2003.
44.
Yoshinori Aono, Yasuhiko Nishioka, K. Mitani, K. Yamabe, M. Inayama, Hiroaki Yanagawa and Saburo Sone : Soluble Fas in Malignant Pleural Effusion and Its Expresion in Lung Cancer Cells, 99th American thoracic society (ATS), Seattle, May 2003.
45.
Saburo Sone : Therapeutic efficacy of new vascular targeting agent, ZD6126 for experimental metastasis of human lung adenocarcinoma in nude mice, The 7th International Symposium on Cancer Chemotherapy, Tokyo, Dec. 2002.
46.
Saburo Sone and Seiji Yano : Molecular target-based therapy against metastsis of lung cancer, International Symposium Molecular Targeting Therapy for Cancer, Hiroshima, Nov. 2002.
47.
Hirohisa Ogawa, Naoki Nishimura, Yasuhiko Nishioka, Masahiko Azuma, Hiroaki Yanagawa and Saburo Sone : Effects of IL-12 gene-transduction into human bronchial epithelial cells on the phenotypic and functional expressions and its therapeutic implication, 7th Congress of Asian Pacific Society of Respirology(APSR), Taiwan, Oct. 2002.
48.
Hiroyuki Taniya, Kenji Tani and Saburo Sone : The expression of chemokine receptor CXCR3 and CCR4 in RA and SLE, 66th American College of Rheumatologyth, New Orleans, Oct. 2002.
49.
Hisatsugu Goto, Seiji Yano, Helong Zhang, Yuka Matsumori, Hirohisa Ogawa, David C. Blakey and Saburo Sone : Activity of a new vascular targeting agent, ZD6126, in pulmonary metastases from human lung adenocarcinoma in nude mice, th International Congress of the Metastasis Research Society, Chicago, Sep. 2002.
50.
Yasuhiko Nishioka, J. Kishi, J. Miyata, M. Azuma, Kenji Tani and Saburo Sone : Macrophage-derived chemokine and thymus and activation-regulated chemokine in bronchoalveolar lavage fluid of patients with interstitial lung diseases, 26th International congress of Internal Medicine, Kyoto, May 2002.
51.
Yasuhiko Nishioka, J. Miyata, Jun Kishi, S. Kakiuchi, Masahiko Azuma, M. Miki, Y. Nakamura, F. Ogushi, T. Sano, H. Bando, Kenji Tani and Saburo Sone : Specific elevation of Th2-type chemokines, MDC and TARC, in bronchoalveolar lavage fluid from patients with eosinophilic pneumonia, 98th American thoracic society(ATS), Atlanta, May 2002.
52.
Jun Kishi, L. Huang, Yasuhiko Nishioka, J. Miyata, F. Ogushi, Kenji Tani and Saburo Sone : Thrombin promotes fibroblast proliteration in experimental radiation pneumonitis, 98th American thoracic society (ATS), Atlanta, May 2002.
53.
Masato Okamoto, S Furuichi, Yasuhiko Nishioka, T Oshikawa, G Ohe, H Nishikawa, T Tano, U S Armed, Y Moriya, T Matsubara, M Saito, Saburo Sone and Mitsunobu Sato : Maturation and activation of human dendritic cells in vitro by lipoteichoic acid-related molecule isolated from OK-432, a streptococcal agent: Involvement of Toll-like receptor 4, American Association for Cancer Research (AAGR), USA, May 2002.
54.
Saburo Sone : Up-to-date cancer treatment -Lung cancer-, The UICC-JSCO joint Fukuoka Cancer Symposium, Fukuoka, Mar. 2002.
55.
Toyokazu Miki, Seiji Yano, Takanori Kanematsu and Saburo Sone : Bone metastasis model with multi-organ dissemination of human small cell lung cancer(SBC-5)cells, 3rd International Conference on Cancer-Induced Bone Diseases, Awaji, Nov. 2001.
56.
Saburo Sone : EGFR tyrosine kinase inhibitor ZD1839, The 6th international Symposium on Cancer Chemotherapy. New Antitumor Agents under Development in the US,Europe and Japan, Tokyo, Nov. 2001.
57.
Yasuhiko Nishioka, N. Nishimura, H. Tahara and Saburo Sone : Intratumoral injection of dendritic cells genetically modified to express interleukin-12 inhibits spontaneous metastases od Lewis lung carcinoma, 97th American thoracic society(ATS), San Francisco, May 2001.
58.
Hirohisa Ogawa, Naoki Nishimura, Yoichi Nakamura, Yasuhiko Nishioka, Masahiko Azuma, Mari Miki, Fumitaka Ogushi and Saburo Sone : Functionl analysis of human bronchial epithelial cell transduced with IL-12 gene by adenovirus vector, American thoracic society (ATS) 2001 97th International Conference, San Francisco, May 2001.
59.
Hirohisa Ogawa, Naoki Nishimura, Yoichi Nakamura, Yasuhiko Nishioka, Masahiko Azuma, Mari Miki, Fumitaka Ogushi and Saburo Sone : Functionl analysis of human bronchial epithelial cell transduced with IL-12 gene by adenovirus vector, American thoracic society (ATS) 2001 97th International Conference, San Francisco, May 2001.
60.
Naoki Nishimura, Yasuhiko Nishioka and Saburo Sone : human dendritic cells transduced with interleukin-12 gene enhanced T cell activation:Centrifugal enhancement of the efficacy of adenovirus-mediated gene transduction, 99nd Annual Meeting of The American Association for Cancer Research, New Orleans, Mar. 2001.
61.
N. Nishimura, Yasuhiko Nishioka, Tsutomu Shinohara and Saburo Sone : Centrifugal enhancement of adenovirus-mediated gene transduction into human dendritic cells, American society for gene therapy(ASGT), Denver, Jun. 2000.
62.
Yasuhiko Nishioka, N. Nishimura, P.D. Robbins, M.T. Lotze, Saburo Sone and H. Tahara : Intratumoral injection of dendritic cells genetically modified to express interleukin-12 induces antitumor immunity and inhibits established tumor growth and spontaneous metastases, American society for gene therapy(ASGT), Denver, Jun. 2000.
63.
H. Ogawa, N. Nishimura, Y. Nakamura, Yasuhiko Nishioka, M. Azuma, M. Miki, F. Ogushi and Saburo Sone : Function analysis of human bronchial epithelial cell transduced with IL-12 gene by adenovirus vector, 96th American thoracic society (ATS), San Francisco, May 2000.
64.
K. Sugihara, Saburo Sone, Masayuki Shono, M. Munekawa, K. Okumura and T. Ogura : Requlation of perforin mRNA expression in CD8+ Cell by monocytes, USA, May 1991.
(キーワード)
Monocyte
65.
Itsuo Katoh, Yoshiyasu Egawa, Tetsuya Kitagawa, Hiroki Taki, Masanori Yoshizumi, Saburo Sone, Hiroaki Yanagawa, Takeshi Ogura, Atsushi Satoh and Yasuro Natori : Intraarterial lymphocyte injection therapy for lymphedema of the extremities, 12th International Congress of Lymphology, Tokyo Kyoto, Aug. 1989.
Le T. Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Souji Kakiuchi, Hisatsugu Goto, Kuramoto Takuya, Mitsuhashi Atsushi, Saburo Sone, Yasuhiko Nishioka and Toyomasa Katagiri : LBMTF a novel transcriptional factor involved in lung cancer bone metastases, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
Atsushi Mitsuhashi, Hisatsugu Goto, Takuya Kuramoto, Syo Tabata, Hirohisa Ogawa, Hisanori Uehara, Shin-ichi Akiyama, Jo Rae Wright, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in nude mice via modulating host immune response., 第52回日本呼吸器学会学術講演会, Apr. 2012.
Yasuhiko Nishioka, Yukinari Kato, Masaki Hanibuchi and Saburo Sone : Antimumor effects of anti-podoplanin antibody NZ-1 against malignant mesothelioma, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
24.
Sho Tabata, Masatatsu Yamamoto, Ryuji Ikeda, Tatsuhiko Furukawa, Tatsuya Kuramoto, Yasuhiko Nishioka, Saburo Sone and Shin-ichi Akiyama : Thymidine-derived sugars generated by thymidine phosphorylase entered glycolytic pathway, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
25.
Le T. Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Takuya Kuramoto, Masaki Hanibuchi, Hisatsugu Goto, Souji Kakiuchi, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Genome-wide gene expression profiling analysis of bone metastases of human non-smal cell lung cancer (NSCLC) in mice, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
Dat LeTan, Taisuke Matsuo, Hisatsugu Goto, Souji Kakiuchi, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Identification and characterization of a novel transcription factor invoved lung cancer bone metastases, 第15回日本がん分子標的治療学会学術集会, Jun. 2011.
Wang Wei, Li Qi, Yamada Tadaaki, Yasuhiko Nishioka, Saburo Sone and Yano Seiji : Stromal fibroblasts induce resistance of lung cancer to EGFR tyrosine kinase inhibitors., 第50回日本呼吸器学会学術講演会English Mini-symposium, Apr. 2010.
Li Qi, Wang Wei, Yamada Tadaaki, Yasuhiko Nishioka and Saburo Sone : Crosstalk straomal fibroblasts promotes malignant pleural mesothelioma progression., 第50回日本呼吸器学会学術講演会English Mini-symposium, Apr. 2010.
75.
Souji Kakiuchi, Yano Seiji, Ogino Hirokazu, Sato Seidai, Hiroya Tada, Yasuhiko Nishioka and Saburo Sone : Novel therapeutic efficacy of E7080 for controlling experimental metastases of human lung cancer cells in natural killer cell-depleted SCID mice., 第50回日本呼吸器学会学術講演会English Mini-symposium, Apr. 2010.
Mitsuhashi Atsushi, Hisatsugu Goto, Kuramoto Takuya, Tabata Syo, Ogawa Hirohisa, Uehara Hisanori, Akiyama Shin-ichi, Wright Rae Jo, Saburo Sone and Yasuhiko Nishioka : Surfactant protein A suppresses progression of human lung adenocarcinoma in nude mice via modulating host immune response., 2011感染免疫クラスター・ミニリトリート, Dec. 2011.
2.
後東 久嗣, Gabr Gomaa Mohammed Adel, 埴淵 昌毅, 倉本 卓哉, 小川 博久, 田畑 祥, Van The Trung, 柿内 聡司, 秋山 伸一, 曽根 三郎, 西岡 安彦 : ヒト肺癌転移モデルにおけるerlotinibの骨転移抑制効果の検討．, 第14回癌と骨病変研究会, 2011年11月.
3.
三橋 惇志, 後東 久嗣, 倉本 卓哉, 柿内 聡司, 佐藤 正大, Van The Trung, 西條 敦郎, 田畑 祥, 小川 博久, 上原 久典, 秋山 伸一, 西岡 安彦, Wright Rae Jo, 曽根 三郎 : 肺サーファクタント蛋白SP-A肺がん進展における機能解析．, 日本肺サーファクタント・界面医学会第47回学術研究会, 2011年10月.