Akiko Abe, Masao Yuasa, Yoshie Imai, Tomohiro Kagawa, Ayuka Mineda, Masato Nishimura, Chisato Tonoiso, Akiko Kubo, Takashi Kawanaka, Hitoshi Ikushima and Takeshi Iwasa : Extreme leanness, lower skeletal muscle quality, and loss of muscle mass during treatment are predictors of poor prognosis in cervical cancer treated with concurrent chemoradiation therapy, International Journal of Clinical Oncology, Vol.27, No.5, 983-991, 2022.
(要約)
Human papillomavirus vaccination is not widespread in Japan, and the low screening rates result in many cases of locally advanced cervical cancer. We investigated the prognostic significance of sarcopenia in patients with cervical cancer to guide healthcare policies to improve treatment outcomes. This retrospective study included 83 patients with cervical cancer without distant metastasis who underwent primary concurrent chemoradiotherapy between 2013 and 2018. We analyzed the indicators of physical condition and muscle quantity using the SYNAPSE VINCENT software. Muscle mass and the relationship between treatment toxicity and prognosis were evaluated. The patients' median age was 60 (range 33-80) years. Cancer stage distribution was as follows: cT2b or higher, 84.3%; N1, 65.1%; and MA, 27.7%. The overall sarcopenia (skeletal muscle index [SMI] < 38.5) rate was 30.1%, and the rate was 33.9 and 22.2% in patients aged < 64 and ≥ 65 years, respectively. No correlation was observed between clinical stage and musculoskeletal indices. Treatment resulted in decreased body weight and SMI; after treatment, the sarcopenia rate increased to 37.3%. A higher intramuscular adipose tissue content (IMAC) reduced the number of chemotherapy cycles needed. Treatment-associated SMI decreases of ≥ 7% indicated poor prognosis, with significant differences in progression-free survival and overall survival (p = 0.013 and p = 0.012, respectively). Patients who were very lean (body mass index < 18.5 kg/m) before treatment had a poor prognosis (p = 0.016 and p < 0.001). Our findings emphasize the importance of assessing original nutritional status and maintaining muscle mass and quality during the treatment of patients with cervical cancer.
Ju-Won Roh, Jung Eun Choi, Hee Dong Han, Wei Hu, Koji Matsuo, Masato Nishimura, Ju-Seog Lee, Sun Young Kwon, Chi Heum Cho, Jongseung Kim, Robert L. Coleman, Gabriel Lopez-Bernstein and Anil K. Sood : Clinical and biological significance of expression in endometrial cancer., Cancer Biology & Therapy, Vol.21, No.2, 147-156, 2020.
(要約)
The objective of this study was to examine the clinical significance of expression and the therapeutic efficacy of its silencing in endometrial cancer. expression in clinical samples was evaluated using a tissue microarray and correlated with clinical outcomes. The biological roles of EZH2 were assayed and . Gene expression was examined to reveal the molecular mechanism underlying the roles of EZH2 in endometrial cancer. We found that overexpression was significantly correlated with disease-free and overall survival of patients with endometrial cancer. silencing resulted in decreased cell viability and invasiveness, and increased apoptosis. In addition, silencing enhanced the cytotoxicity of taxanes and cisplatin in Hec-1A and Ishikawa endometrial cancer cells. silencing using small-interfering RNA (siRNA) incorporated into chitosan nanoparticles (siRNA/CN) induced a significant anti-tumor effect compared with that observed in controls (66.6% reduction in Hec-1A cells and 63.2% reduction in Ishikawa cells, < .05 for both). Moreover, siRNA/CN in combination with taxanes produced more robust anti-tumor effects versus those induced by monotherapies (77.0% for Hec-1A cells and 57.7% for Ishikawa cells, < .05 for both). These results were associated with decreased angiogenesis and cell proliferation, and enhanced apoptosis. Genomic analyses revealed that silencing decreased the expression levels of many genes associated with tumor growth, including . Collectively, these results support as an attractive target for the therapeutic management of endometrial cancer.
Ayuka Mineda, Masato Nishimura, Tomohiro Kagawa, Eri Takiguchi, Takako Kawakita, Akiko Abe and Minoru Irahara : Resveratrol suppresses proliferation and induces apoptosis of uterine sarcoma cells by inhibiting the Wnt signaling pathway., Experimental and Therapeutic Medicine, Vol.17, No.3, 2242-2246, 2019.
(要約)
Resveratrol, a natural product and peroxisome proliferator-activated receptor (PPAR) agonist, has been reported to exert anti-cancer effects in several tumor models. A previous study by our group reported that prostaglandin J2, a PPARγ ligand, inhibited cell proliferation in a uterine sarcoma cell line. The aim of the present study was to investigate the role of the Wnt signaling pathway in resveratrol-induced apoptosis and inhibition of cell proliferation in the MES-SA human uterine sarcoma cell line. A WST-1 assay demonstrated that resveratrol inhibited cell proliferation in the MES-SA cell line, and flow cytometry revealed that the number of apoptotic cells increased in a resveratrol dose-dependent manner. The mechanisms underlying these effects of resveratrol were speculated to involve the expression of β-catenin and its target gene, c-myc, which were examined using western blot analysis. The results revealed a dose-dependent downregulation of this β-catenin and c-myc. This effect was blunted by a pharmacological inhibitor of glycogen synthase kinase 3β. Therefore, it is likely that resveratrol inhibited the cell proliferation and increased the number of apoptotic cells, at least partially, via the Wnt signaling pathway. The present results suggest that resveratrol is a potential candidate for the treatment of uterine sarcoma.
Kanako Yoshida, Masato Nishimura, Akiko Abe, Takeshi Katou, Hiroyuki Furumoto and Minoru Irahara : Can systematic lymphadenectomy be omitted for low-risk endometrial cancer?, The Journal of Medical Investigation : JMI, Vol.65, No.3,4, 221-224, 2018.
(要約)
The objective of this study was to identify pathological indicators that could be used to identify a subgroup of patients with apparent stage I endometrial cancer who do require retroperitoneal lymphadenectomy. 188 T1 endometrial cancer patients underwent primary surgery at Tokushima University Hospital. We retrospectively evaluated their clinical records and histopathological factors. Systematic lymphadenectomy was performed for 149 patients, and 39 patients (grade 1 with < 5 mm of myometrial invasion) were treated without lymphadenectomy. Lymph node metastases were found in 19 (12.8%) of the lymphadenectomy cases. Twenty-four patients with a T1a endometrium-limited lesion did not exhibit lymph node metastasis. Three (3.1%) of the 95 patients with a T1a lesion exhibited lymph node metastasis, and these 3 cases exhibited approximately 50% myometrial invasion. The 39 low-risk patients who did not undergo systematic lymphadenectomy remain alive without recurrence. Systematic lymphadenectomy could be omitted for patients with a grade 1 tumor and minor myometrial invasion of less than 5mm. J. Med. Invest. 65:221-224, August, 2018.
Mayumi Takeuchi, Kenji Matsuzaki, Yoshimi Bando, Masato Nishimura, Aki Hayashi and Masafumi Harada : A Case of Uterine Tumor Resembling Ovarian Sex-cord Tumor (UTROSCT) Exhibiting Similar Imaging Characteristics to Those of Ovarian Sex-cord Tumor., Magnetic Resonance in Medical Sciences, Vol.18, No.2, 113-114, 2018.
Akiko Abe, Masato Nishimura and Minoru Irahara : See and treat LEEP biopsy for cervical intraepithelial neoplasia grade 2/3, European Journal of Gynaecological Oncology, Vol.39, No.6, 958-962, 2018.
Eri Takiguchi, Masato Nishimura, Ayuka Mineda, Takako Kawakita, Akiko Abe and Minoru Irahara : Growth inhibitory effect of the Src inhibitor dasatinib in combination with anticancer agents on uterine cervical adenocarcinoma cells., Experimental and Therapeutic Medicine, Vol.14, No.5, 4293-4299, 2017.
(要約)
Uterine cervical adenocarcinoma has a poor clinical prognosis when compared with squamous cell carcinoma. Therefore, the development of new treatment strategies for uterine cervical adenocarcinoma is necessary. Src is a proto-oncogene that is important in cancer progression. Dasatinib is a Src inhibitor that has been reported to be effective when used in combination with anticancer drugs. The present study aimed to confirm Src expression in human cervical adenocarcinoma cell lines and to determine the mechanism underlying the inhibitory effect of dasatinib on Src signaling . Western blot analysis was performed to investigate Src expression in cervical adenocarcinoma cell lines (HeLa and TCO-2 cells). The cells were cultured for 48 h with the addition of different concentrations of anticancer drugs (paclitaxel or oxaliplatin). Viable cell count was measured using a colorimetric (WST-1) assay. The concentrations of anticancer agents were fixed according to the results obtained, and the same experiments were performed using the drugs in combination with dasatinib at various concentrations to determine the concentrations that significantly affected the number of viable cells. The presence or absence of apoptosis was investigated using a caspase-3/7 assay. Signal transduction in each cell line was examined using western blotting. Src was activated in the two cell lines, and cell proliferation was significantly suppressed by each anticancer drug in combination with 10 µM dasatinib. Caspase-3/7 activity was also increased and Src signaling was suppressed by each anticancer drug in combination with dasatinib. In conclusion, Src is overexpressed in cervical adenocarcinoma cell lines, and dasatinib inhibits intracellular Src signaling and causes apoptosis. The results of the present study suggest that Src may be targeted in novel therapeutic strategies for cervical adenocarcinoma.
Mayumi Takeuchi, Kenji Matsuzaki, Koichi Tsuneyama, Masato Nishimura, Eri Takiguchi and Masafumi Harada : Ovarian Large Cell Neuroendocrine Carcinoma Associated with Serous Carcinoma: Correlation of Pathology with MR Imaging, Magnetic Resonance in Medical Sciences, Vol.16, No.4, 273-274, 2017.
Takako Kawakita, Masato Nishimura, Eri Takiguchi, Akiko Abe and Minoru Irahara : Cytotoxic effects of 15-deoxy-Δ12,14-prostaglandin J2 alone and in combination with dasatinib against uterine sarcoma ., Experimental and Therapeutic Medicine, Vol.13, No.6, 2939-2945, 2017.
(要約)
Effective chemotherapeutic strategies for uterine sarcoma are lacking; existing therapies achieve poor response rates. Previous studies have identified the prostaglandin 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) as a potential anticancer treatment; however, its effectiveness in uterine sarcoma has not been examined. Furthermore, the molecular mechanisms underlying the cytotoxic mechanism of 15d-PGJ2 remain unclear. Here, we evaluated the effects of 15d-PGJ2 alone and in combination with the tyrosine kinas inhibitor (TKI) dasatinib in uterine sarcoma cell lines (MES-SA, MES-SA/DX5 and SKN). 15d-PGJ2 inhibited cell growth and increased apoptosis. Western blotting demonstrated that 15d-PGJ2 treatment increased MEK and ERK phosphorylation, and decreased levels of phosphorylated AKT. Dasatinib in combination with 15d-PGJ2 significantly reduced cell proliferation compared with 15d-PGJ2 alone, and repressed both the AKT and MAPK pathways. The cell growth inhibition rate in the PGJ2 was 21.5±12.0, 35.3±5.4 and 28.3±4.2%, respectively (MES-SA, MES-SA/DX5 and SKN cell lines) and the cell growth inhibition rate in the combination therapy was significantly higher compared with 15d-PGJ2 alone (MES-SA; 64.2±0.8, MES-SA/DX5;23.9±8.2 and SKN; 41.4±17.6%). The PGJ2 IC determined by MTT assay was 27.41,10.46 and 17.38 µmol/l, respectively (MES-SA, MES-SA/DX5 and SKN cell lines) and the dasatinib IC was 6.68,17.30 and 6.25 µmol/l, respectively. Our findings demonstrate that 15d-PGJ2 suppresses proliferation by inactivating the AKT pathway in uterine sarcoma. Furthermore, combining 15d-PGJ2 with dasatinib produced a synergistic effect on cancer cell inhibition by repressing 15d-PGJ2-mediated activation of MAPK signaling, and further repressing AKT signaling. These results suggest that 15d-PGJ2 could be used in combination with dasatinib as a potential therapeutic approach for uterine sarcoma.
K Matsuo, Y Takazawa, M S. Ross, E Elishaev, I Podzielinski, M Yunokawa, T B. Sheridan, S H. Bush, M M. Klobocista, E A. Blake, T Takano, S Matsuzaki, T Baba, S Satoh, M Shida, T Nishikawa, Y Ikeda, S Adachi, T Yokoyama, M Takekuma, K Fujiwara, Y Hazama, D Kadogami, M N. Moffitt, S Takeuchi, Masato Nishimura, K Iwasaki, N Ushioda, M S. Johnson, M Yoshida, A Hakam, S W. Li, A M. Richmond, H Machida, P Mhawech-Fauceglia, Y Ueda, K Yoshino, K Yamaguchi, T Oishi, H Kajiwara, K Hasegawa, M Yasuda, K Kawana, K Suda, T M. Miyake, T Moriya, Y Yuba, T Morgan, T Fukagawa, A Wakatsuki, T Sugiyama, T Pejovic, T Nagano, K Shimoya, M Andoh, Y Shiki, T Enomoto, T Sasaki, K Fujiwara, M Mikami, M Shimada, I Konishi, T Kimura, M D. Post, M M. Shahzad, D D. Im, H Yoshida, K Omatsu, F R. Ueland, J L. Kelley, R G. Karabakhtsian and L D. Roman : Significance of histologic pattern of carcinoma and sarcoma components on survival outcomes of uterine carcinosarcoma., Annals of Oncology, Vol.27, No.7, 1257-1266, 2016.
(要約)
To examine the effect of the histology of carcinoma and sarcoma components on survival outcome of uterine carcinosarcoma. A multicenter retrospective study was conducted to examine uterine carcinosarcoma cases that underwent primary surgical staging. Archived slides were examined and histologic patterns were grouped based on carcinoma (low-grade versus high-grade) and sarcoma (homologous versus heterologous) components, correlating to clinico-pathological demographics and outcomes. Among 1192 cases identified, 906 cases were evaluated for histologic patterns (carcinoma/sarcoma) with high-grade/homologous (40.8%) being the most common type followed by high-grade/heterologous (30.9%), low-grade/homologous (18.0%), and low-grade/heterologous (10.3%). On multivariate analysis, high-grade/heterologous (5-year rate, 34.0%, P = 0.024) and high-grade/homologous (45.8%, P = 0.017) but not low-grade/heterologous (50.6%, P = 0.089) were independently associated with decreased progression-free survival (PFS) compared with low-grade/homologous (60.3%). In addition, older age, residual disease at surgery, large tumor, sarcoma dominance, deep myometrial invasion, lymphovascular space invasion, and advanced-stage disease were independently associated with decreased PFS (all, P < 0.01). Both postoperative chemotherapy (5-year rates, 48.6% versus 39.0%, P < 0.001) and radiotherapy (50.1% versus 44.1%, P = 0.007) were significantly associated with improved PFS in univariate analysis. However, on multivariate analysis, only postoperative chemotherapy remained an independent predictor for improved PFS [hazard ratio (HR) 0.34, 95% confidence interval (CI) 0.27-0.43, P < 0.001]. On univariate analysis, significant treatment benefits for PFS were seen with ifosfamide for low-grade carcinoma (82.0% versus 49.8%, P = 0.001), platinum for high-grade carcinoma (46.9% versus 32.4%, P = 0.034) and homologous sarcoma (53.1% versus 38.2%, P = 0.017), and anthracycline for heterologous sarcoma (66.2% versus 39.3%, P = 0.005). Conversely, platinum, taxane, and anthracycline for low-grade carcinoma, and anthracycline for homologous sarcoma had no effect on PFS compared with non-chemotherapy group (all, P > 0.05). On multivariate analysis, ifosfamide for low-grade/homologous (HR 0.21, 95% CI 0.07-0.63, P = 0.005), platinum for high-grade/homologous (HR 0.36, 95% CI 0.22-0.60, P < 0.001), and anthracycline for high-grade/heterologous (HR 0.30, 95% CI 0.14-0.62, P = 0.001) remained independent predictors for improved PFS. Analyses of 1096 metastatic sites showed that carcinoma components tended to spread lymphatically, while sarcoma components tended to spread loco-regionally (P < 0.001). Characterization of histologic pattern provides valuable information in the management of uterine carcinosarcoma.
Mayumi Takeuchi, Kenji Matsuzaki, Yoshimi Bando, Masato Nishimura, Akiko Yoneda and Masafumi Harada : A case of polypoid endometriosis with malignant transformation., Abdominal Radiology, Vol.41, No.9, 1699-1702, 2016.
(要約)
Polypoid endometriosis is a benign, rare variant of endometriosis which forms multiple polypoid nodules in the female pelvis mimicking malignant tumors; however, it may rarely cause malignant transformation. We report magnetic resonance imaging findings of a case of polypoid endometriosis with malignant transformation. Multiple high-signal intensity polypoid nodules in the cul-de-sac surrounded by low-signal intensity rim-like fibrous adhesion protruding to the posterior wall of the uterine body were demonstrated on T2-weighted images. The polypoid nodules showed weak contrast enhancement compared with that of uterine myometrium on post-contrast T1-weighted images, and slight high signal intensity on diffusion-weighted images with relatively high mean apparent diffusion coefficient. Reported cases of polypoid endometriosis showed intense contrast enhancement similar to that of uterine myometrium, and weak contrast enhancement similar to that of endometrial carcinoma may be suggestive for malignant transformation of polypoid endometriosis.
Yasuyo Saijo, Hiroyuki Furumoto, Kanako Yoshida, Masato Nishimura and Minoru Irahara : Clinical Significance of Vascular Endothelial Growth Factor Expression and Microvessel Density in Invasive Cervical Cancer., The Journal of Medical Investigation : JMI, Vol.62, No.3-4, 154-160, 2015.
(要約)
To determine whether vascular endothelial growth factor (VEGF) expression and microvessel density are predictive of prognosis in cases of invasive cervical cancer, correlations among VEGF expression, microvessel density, and clinicopathological parameters were identified. VEGF expression was evaluated in 50 cervical cancer samples by immunohistochemical staining. Microvessel density was assessed by immunostaining for CD31-positive endothelial cells in the most vascularized areas of tumors. VEGF expression and microvessel density were significantly higher in adenocarcinomas than in squamous cell carcinomas. However, in cases of adenocarcinoma, no significant correlations were found among VEGF expression, microvessel density, and clinicopathological parameters. In contrast, for squamous cell carcinomas, microvessel density was significantly higher in cases at an advanced stage and in those with several other poor prognostic factors. The finding that cervical adenocarcinomas exhibited greater VEGF expression and microvessel density than squamous cell carcinomas may explain the poorer prognosis of adenocarcinoma compared with squamous cell carcinoma. Moreover, microvessel density in squamous cell carcinomas was significantly correlated with poor prognostic factors. Therefore, there is possibility that bevacizumab, a humanized monoclonal antibody against VEGF-A, may be useful in the initial treatment targeting angiogenesis for early-stage cervical cancer.
Kanako Yoshida, Hiroyuki Furumoto, Akiko Abe, Takeshi Katou, Masato Nishimura and Minoru Irahara : The possibility of vertical transmission of human papillomavirus through maternal milk, Journal of Obstetrics and Gynaecology, Vol.31, No.6, 503-506, 2011.
(要約)
Human papillomavirus (HPV) DNA has been detected in the oral cavity of infants and breast cancer tissue, suggesting its vertical transmission through maternal milk. We determined whether HPV is detected in maternal milk and is vertically transmitted by breast-feeding. Informed consent was obtained, and maternal milk samples (n=80) were analysed for high-risk HPV DNA. In 43 women, this DNA was measured in the uterine cervix. In women with positive samples, this DNA was measured in the oral cavities of their children. The domain including HPV E6 and E7 was amplified by polymerase chain reaction using consensus primers, and HPV serotype determined by electrophoresis after restriction enzyme digestion. High-risk HPV-16 was detected in two of 80 samples (2.5%), and in these two cases, high-risk HPV was not detected in the uterine cervix or oral cavity of the child. It was concluded that the infection of HPV in maternal milk is rare (2/80); vertical transmission through maternal milk was not detected in this study (0/80). HPV infection through maternal milk may occur, but its likelihood is low.
Wei Hu, Chunhua Lu, Han Hee Dong, Jie Huang, De-yu Shen, Rebecca L. Stone, Alpa M. Nick, Mian M. K. Shahzad, Edna Mora, Nicholas B. Jennings, Sun Joo Lee, Ju-Won Roh, Koji Matsuo, Masato Nishimura, Blake W. Goodman, Robert B. Jaffe, Robert R. Langley, Michael T. Deavers, Gabriel Lopez-Berestein, Robert L. Coleman and Anil K. Sood : Biological roles of the Delta family Notch ligand Dll4 in tumor and endothelial cells in ovarian cancer., Cancer Research, Vol.71, No.18, 6030-6039, 2011.
(要約)
Emerging evidence suggests that the Notch/Delta-like ligand 4 (Dll4) pathway may offer important new targets for antiangiogenesis approaches. In this study, we investigated the clinical and biological significance of Dll4 in ovarian cancer. Dll4 was overexpressed in 72% of tumors examined in which it was an independent predictor of poor survival. Patients with tumors responding to anti-VEGF therapy had lower levels of Dll4 than patients with stable or progressive disease. Under hypoxic conditions, VEGF increased Dll4 expression in the tumor vasculature. Immobilized Dll4 also downregulated VEGFR2 expression in endothelial cells directly through methylation of the VEGFR2 promoter. RNAi-mediated silencing of Dll4 in ovarian tumor cells and tumor-associated endothelial cells inhibited cell growth and angiogenesis, accompanied by induction of hypoxia in the tumor microenvironment. Combining Dll4-targeted siRNA with bevacizumab resulted in greater inhibition of tumor growth, compared with control or treatment with bevacizumab alone. Together, our findings establish that Dll4 plays a functionally important role in both the tumor and endothelial compartments of ovarian cancer and that targeting Dll4 in combination with anti-VEGF treatment might improve outcomes of ovarian cancer treatment.
Koji Matsuo, Masato Nishimura, Justin N. Bottsford-Miller, Jie Huang, Kakajan Komurov, Guillermo N. Armaiz-Pena, Mian M. K. Shahzad, Rebecca L. Stone, Ju Won Roh, Angela M. Sanguino, Chunhua Lu, Dwight D. Im, Neil B. Rosenshien, Atsuko Sakakibara, Tadayoshi Nagano, Masato Yamasaki, Takayuki Enomoto, Tadashi Kimura, Prahlad T. Ram, Kathleen M. Schmeler, Gary E. Gallick, Kwong K. Wong, Michael Frumovitz and Anil K. Sood : Targeting SRC in mucinous ovarian carcinoma., Clinical Cancer Research, Vol.17, No.16, 5367-5378, 2011.
(要約)
Mucinous ovarian carcinomas have a distinct clinical pattern compared with other subtypes of ovarian carcinoma. Here, we evaluated (i) stage-specific clinical significance of mucinous ovarian carcinomas in a large cohort and (ii) the functional role of Src kinase in preclinical models of mucinous ovarian carcinoma. A total of 1,302 ovarian cancer patients including 122 (9.4%) cases of mucinous carcinoma were evaluated for survival analyses. Biological effects of Src kinase inhibition were tested using dasatinib-based therapy in a novel orthotopic mucinous ovarian cancer model (RMUG-S-ip2). Patients with advanced-stage mucinous ovarian cancer had significantly worse survival than those with serous histology: median overall survival, 1.67 versus 3.41 years, P = 0.002; median survival time after recurrence of 0.53 versus 1.66 years, P < 0.0001. Among multiple ovarian cancer cell lines, RMUG-S-ip2 mucinous ovarian cancer cells showed the highest Src kinase activity. Moreover, oxaliplatin treatment induced phosphorylation of Src kinase. This induced activity by oxaliplatin therapy was inhibited by concurrent administration of dasatinib. Targeting Src with dasatinib in vivo showed significant antitumor effects in the RMUG-S-ip2 model but not in the serous ovarian carcinoma (SKOV3-TR) model. Combination therapy of oxaliplatin with dasatinib further showed significant effects on reducing cell viability, increasing apoptosis, and in vivo antitumor effects in the RMUG-S-ip2 model. Our results suggest that poor survival of women with mucinous ovarian carcinoma is associated with resistance to cytotoxic therapy. Targeting Src kinase with a combination of dasatinib and oxaliplatin may be an attractive approach for this disease.
Hee Dong Han, Edna M. Mora, Ju Won Roh, Masato Nishimura, Sun Joo Lee, Rebecca L. Stone, Menashe Bar-Eli, Gabriel Lopez-Berestein and Anil K. Sood : Chitosan hydrogel for localized gene silencing., Cancer Biology & Therapy, Vol.11, No.9, 839-845, 2011.
(要約)
To achieve effective delivery of siRNA into target cells in vivo, we have developed a novel approach of siRNA delivery by using local drug delivery systems. The chitosan hydrogel (CH-HG) displayed a liquid-solid phase transition in a temperature-dependent manner and formed an endothermic hydrogel in tumor tissue after intra-tumoral injection. Additionally, we tested the extent of in vivo delivery following a single intra-tumoral injection of Alexa555 siRNA/CH-HG into A375SM-bearing mice. The Alexa555 siRNA demonstrated higher localization into tumor cells compared to control. The Alexa555 siRNA delivery extends to tumor cells outside of CH-HG and some tumor cells also infiltrated into CH-HG. For therapeutic proof-of-concept studies, CH-HG including TG2-targeted siRNA significantly inhibited tumor growth in melanoma (A375SM) and breast (MDA-MB231) tumor models compared to control (A375SM: 72% reduction and MDA-MB231: 92% reduction, p < 0.001). we prepared a CH-HG system loaded with siRNA to enhance localized therapeutic efficacy without risk for systemic side effects. Delivery of siRNA into CH-HG was confirmed by fluorescence microscopy. Antitumor efficacy was examined in mouse models of melanoma (A375SM) and breast (MDA-MD231) cancer. This study developed a novel local delivery method for siRNA therapy using the CH-HG system. This approach could have broad applications for multiple localized diseases.
(キーワード)
Animals / Breast Neoplasms / Cell Line, Tumor / Chitosan / Drug Delivery Systems / Female / Gene Silencing / Humans / Hydrogels / Melanoma / Mice / Mice, Nude / RNA, Small Interfering / Xenograft Model Antitumor Assays
Mian M. K. Shahzad, Lingegowda S. Mangala, Hee Dong Han, Chunhua Lu, Justin Bottsford-Miller, Masato Nishimura, Edna M. Mora, Jeong-Won Lee, Rebecca L. Stone, Chad V. Pecot, Duangmani Thanapprapasr, Ju-Won Roh, Puja Gaur, Maya P. Nair, Yun-Yong Park, Nirupama Sabnis, Michael T. Deavers, Ju-Seog Lee, Lee M. Ellis, Gabriel Lopez-Berestein, Walter J. McConathy, Laszlo Prokai, Andras G. Lacko and Anil K. Sood : Targeted delivery of small interfering RNA using reconstituted high-density lipoprotein nanoparticles., Neoplasia, Vol.13, No.4, 309-319, 2011.
(要約)
RNA interference holds tremendous potential as a therapeutic approach, especially in the treatment of malignant tumors. However, efficient and biocompatible delivery methods are needed for systemic delivery of small interfering RNA (siRNA). To maintain a high level of growth, tumor cells scavenge high-density lipoprotein (HDL) particles by overexpressing its receptor: scavenger receptor type B1 (SR-B1). In this study, we exploited this cellular characteristic to achieve efficient siRNA delivery and established a novel formulation of siRNA by incorporating it into reconstituted HDL (rHDL) nanoparticles. Here, we demonstrate that rHDL nanoparticles facilitate highly efficient systemic delivery of siRNA in vivo, mediated by the SR-B1. Moreover, in therapeutic proof-of-concept studies, these nanoparticles were effective in silencing the expression of two proteins that are key to cancer growth and metastasis (signal transducer and activator of transcription 3 and focal adhesion kinase) in orthotopic mouse models of ovarian and colorectal cancer. These data indicate that an rHDL nanoparticle is a novel and highly efficient siRNA carrier, and therefore, this novel technology could serve as the foundation for new cancer therapeutic approaches.
Hye-Sun Kim, Hee Dong Han, Guillermo N. Armaiz-Pena, Rebecca L. Stone, Eun Ji Nam, Jeong-Won Lee, Mian M. K. Shahzad, Alpa M. Nick, Sun Joo Lee, Ju-Won Roh, Masato Nishimura, Lingegowda S. Mangala, Justin Bottsford-Miller, Gary E. Gallick, Gabriel Lopez-Berestein and Anil K. Sood : Functional roles of Src and Fgr in ovarian carcinoma., Clinical Cancer Research, Vol.17, No.7, 1713-1721, 2011.
(要約)
Src is an attractive target because it is overexpressed in a number of malignancies, including ovarian cancer. However, the effect of Src silencing on other Src family kinases (SFKs) is not known. We hypothesized that other SFK members could compensate for the lack of Src activity. Cell viability after either Src or Fgr silencing was examined in ovarian cancer cell lines by MTT assay. Expression of SFKs after Src silencing in ovarian cancer cells was examined by real-time reverse transcriptase (RT)-PCR. Therapeutic effect of in vivo Src and/or Fgr silencing was examined using siRNA incorporated into chitosan nanoparticles (siRNA/CH-NP). Microvessel density, cell proliferation, and apoptosis markers were determined by immunohistochemical staining in ovarian tumor tissues. Src silencing enhanced cytotoxicity of docetaxel in both SKOV3ip1 and HeyA8 cells. In addition, Src silencing using siRNA/CH-NP in combination with docetaxel resulted in significant inhibition of tumor growth compared with control siRNA/CH-NP (81.8% reduction in SKOV3ip1, P = 0.017; 84.3% reduction in HeyA8, P < 0.005). These effects were mediated by decreased tumor cell proliferation and angiogenesis, and increased tumor cell apoptosis. Next, we assessed the effects of Src silencing on other SFK members in ovarian cancer cell lines. Src silencing resulted in significantly increased Fgr levels. Dual Src and Fgr silencing in vitro resulted in increased apoptosis that was mediated by increased caspase and AKT activity. In addition, dual silencing of Src and Fgr in vivo using siRNA/CH-NP resulted in the greatest reduction in tumor growth compared with silencing of either Src or Fgr alone in the HeyA8 model (68.8%, P < 0.05). This study demonstrates that, in addition to Src, Fgr plays a biologically significant role in ovarian cancer growth and might represent an important target.
MM Shahzad, JM Arevalo, GN Armaiz-Pena, C Lu, RL Stone, M Moreno-Smith, Masato Nishimura, JW Lee, NB Jennings, J Bottsford-Miller, P Vivas-Mejia, SK Lutgendorf, G Lopez-Berestein, M Bar-Eli, SW Cole and AK Sood : Stress effects on FosB- and interleukin-8 (IL8)-driven ovarian cancer growth and metastasis., The Journal of Biological Chemistry, Vol.285, No.46, 35462-35470, 2010.
(要約)
A growing number of studies indicate that chronic stress can accelerate tumor growth due to sustained sympathetic nervous system activation. Our recent findings suggest that chronic stress is associated with increased IL8 levels. Here, we examined the molecular and biological significance of IL8 in stress-induced tumor growth. Norepinephrine (NE) treatment of ovarian cancer cells resulted in a 250-300% increase in IL8 protein and 240-320% increase in its mRNA levels. Epinephrine treatment resulted in similar increases. Moreover, NE treatment resulted in a 3.5-4-fold increase in IL8 promoter activity. These effects were blocked by propranolol. Promoter deletion analyses suggested that AP1 transcription factors might mediate catecholamine-stimulated up-regulation of IL8. siRNA inhibition studies identified FosB as the pivotal component responsible for IL8 regulation by NE. In vivo chronic stress resulted in increased tumor growth (by 221 and 235%; p < 0.01) in orthotopic xenograft models involving SKOV3ip1 and HeyA8 ovarian carcinoma cells. This enhanced tumor growth was completely blocked by IL8 or FosB gene silencing using 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine nanoliposomes. IL8 and FosB silencing reduced microvessel density (based on CD31 staining) by 2.5- and 3.5-fold, respectively (p < 0.001). Our findings indicate that neurobehavioral stress leads to FosB-driven increases in IL8, which is associated with increased tumor growth and metastases. These findings may have implications for ovarian cancer management.
WA Spannuth, LS Mangala, RL Stone, AR Carroll, Masato Nishimura, MM Shahzad, SJ Lee, M Moreno-Smith, AM Nick, R Liu, NB Jennings, YG Lin, WM Merritt, RL Coleman, PE Vivas-Mejia, Y Zhou, V Krasnoperov, G Lopez-Berestein, PS Gill and AK Sood : Converging evidence for efficacy from parallel EphB4-targeted approaches in ovarian carcinoma., Molecular Cancer Therapeutics, Vol.9, No.8, 2377-2388, 2010.
(要約)
EphB4 is a transmembrane receptor tyrosine kinase that plays an important role in neural plasticity and angiogenesis. EphB4 is overexpressed in ovarian cancer and is predictive of poor clinical outcome. However, the biological significance of EphB4 in ovarian cancer is not known and is the focus of the current study. Here, we examined the biological effects of two different methods of EphB4 targeting (a novel monoclonal antibody, EphB4-131 or siRNA) using several ovarian cancer models. EphB4 gene silencing significantly increased tumor cell apoptosis and decreased migration (P < 0.001) and invasion (P < 0.001). Compared with controls, EphB4 siRNA-1,2-dioleoyl-sn-glycero-3-phosphatidylcholine alone significantly reduced tumor growth in the A2780-cp20 (48%, P < 0.05) and IGROV-af1 (61%, P < 0.05) models. Combination therapy with EphB4 siRNA-1,2-dioleoyl-sn-glycero-3-phosphatidylcholine and docetaxel resulted in the greatest reduction in tumor weight in both A2780-cp20 and IGROV-af1 models (89-95% reduction versus controls; P < 0.05 for both groups). The EphB4-131 antibody, which reduced EphB4 protein levels, decreased tumor growth by 80% to 83% (P < 0.01 for both models) in A2780-cp20 and IGROV-af1 models. The combination of EphB4-131 and docetaxel resulted in the greatest tumor reduction in both A2780-cp20 and IGROV-af1 models (94-98% reduction versus controls; P < 0.05 for both groups). Compared with controls, EphB4 targeting resulted in reduced tumor angiogenesis (P < 0.001), proliferation (P < 0.001), and increased tumor cell apoptosis (P < 0.001), which likely occur through modulation of phosphoinositide 3-kinase signaling. Collectively, these data identify EphB4 as a valuable therapeutic target in ovarian cancer and offer two new strategies for further development.
Masahiro Murakami, Hisanori Uehara, Masato Nishimura, Takeshi Iwasa and Hiroshi Ikawa : A huge ovarian smooth muscle tumor: a case report., The Journal of Medical Investigation : JMI, Vol.57, No.1-2, 158-162, 2010.
(要約)
Ovarian smooth muscle tumors are a very rare type of ovarian tumor. In this paper, we report the case of a 62-year-old woman who had a huge smooth muscle tumor of the right ovary. The values of all the serum tumor markers were within normal limit. The tumor measured 25 cm in diameter and weighed 6,200 g. Histological examination revealed that coagulative cellular atypia was moderate to severe, necrosis was not present and mitotic index was low. According to the criteria for the evaluation of the uterine smooth muscle tumors, this huge tumor was diagnosed as atypical leiomyoma. However, we finally made a diagnosis of this tumor as a smooth muscle tumor of uncertain malignant potential (STUMP) because of its huge size. Further information is required regarding the characteristics of ovarian smooth muscle tumor and the propriety to introduce uterine tumor histological criteria to ovarian tumors.
Koji Matsuo, Malcolm S. Ross, Mayu Yunokawa, Marian S. Johnson, Hiroko Machida, Kohei Omatsu, Merieme M. Klobocista, Dwight D. Im, Shinya Satoh, Tsukasa Baba, Yuji Ikeda, Stephen H. Bush, Kosei Hasegawa, Erin A. Blake, Munetaka Takekuma, Masako Shida, Masato Nishimura, Sosuke Adachi, Tanja Pejovic, Satoshi Takeuchi, Takuhei Yokoyama, Yutaka Ueda, Keita Iwasaki, Takahito M. Miyake, Shiori Yanai, Tadayoshi Nagano, Tadao Takano, Mian Mk Shahzad, Frederick R. Ueland, Joseph L. Kelley and Lynda D. Roman : Tumor characteristics and outcome of uterine carcinosarcoma in women aged ≥80 years., Surgical Oncology, Vol.29, 25-32, Feb. 2019.
(要約)
To examine clinico-pathological characteristics and outcomes of uterine carcinosarcoma (UCS) in women aged ≥80 years. This is a secondary analysis of a previous multicenter retrospective study examining 906 women with stage I-IV UCS who underwent primary hysterectomy. Patient demographics, treatment types, tumor characteristics, and survival were examined across aged ≥80 (n = 82 [9.1%]), aged 60-79, (n = 526 [58.1%]), and aged <60 (n = 298 [32.9%]). Women in the aged ≥80 group were more likely to be Caucasian, undergo simple hysterectomy without lymphadenectomy, and receive no postoperative therapy (all, P < 0.05). Tumors in the aged ≥80 group were more likely to have high-grade carcinoma, heterologous sarcoma, and sarcoma dominance but less likely to have lympho-vascular space invasion (all, P < 0.05). Lymphadenectomy did not improve survival in the aged ≥80 group (P > 0.05), whereas lymphadenectomy was protective for survival in the younger groups (both, P < 0.05). Postoperative chemotherapy was associated with improved progression-free survival (PFS) in the aged ≥80 group (hazard ratio [HR] 0.44, 95% confidence interval [CI] 0.22-0.89, P = 0.021). With chemotherapy treatment, women in the aged ≥80 group had PFS similar to those in the aged 60-79 group (HR 0.97, 95%CI 0.51-1.83, P = 0.92). In contrast, without chemotherapy treatment, women in the aged ≥80 group had significantly decreased PFS compared to the aged 60-79 group (HR 1.62, 95%CI 1.09-2.40, P = 0.016). Similar associations were observed for postoperative radiotherapy. Nearly 10% of women with UCS are aged ≥80 that are characterized by aggressive tumor factors. Postoperative therapy but not extensive surgery may improve survival in this age group.
Akiko Abe, Akira Kuwahara, Tomohiro Kagawa, Ayuka Mineda and Masato Nishimura : A survey of germline mutations with epithelial ovarian cancer in Japanese patients., ESGO Annual Meeting 2019, Greece, Nov. 2019.
2.
Chisato Tonoiso, Hitoshi Ikushima, Akihiro Haga, Shunsuke Furutani, Takashi Kawanaka, Akiko Kubo, Masato Nishimura, Akiko Abe, Minoru Irahara and Masafumi Harada : Investigation of prognostic factors of cervical cancer using pre-treatment MRI images, The 2018 International Gynecologic Cancer Society Meeting, Sep. 2018.
3.
Akiko Abe, Masato Nishimura, Eri Takiguchi, Takako Kawakita, TANIGUCHI Yuka, Yuri Yamamoto, YAMASAKI Mikio, Takeshi Iwasa, Akira Kuwahara, Toshiya Matsuzaki and Minoru Irahara : Survey on oncologist and gynecologist concerned with fertility for cancer survivors in reproductive age., IFFS/JSRM International Meeting,2015, Yokohama, Apr. 2015.
4.
Masato Nishimura, Eri Takiguchi, Takako Kawakita, Akiko Abe, TANIGUCHI Yuka, Yuri Yamamoto, YAMASAKI Mikio, Takeshi Iwasa, Akira Kuwahara, Toshiya Matsuzaki and Minoru Irahara : Efficacy and pregnant outcomes of fertility-sparing treatment with medroxyprogesterone acetate for endometrial carcinoma and atypical endometrial hyperplasia in young women., IFFS/JSRM International Meeting,2015, Yokohama, Apr. 2015.
竹内 麻由美, 松崎 健司, 坂東 良美, 西村 正人, 原田 雅史 : 子宮内膜症関連腫瘍として発生した卵巣漿液粘液性境界悪性腫瘍と癌の併存症例:発生機序及びMRIと病理の対比検討, JSAWI (The Japanese Society for the Advancement of Women's Imaging) 第24回シンポジウム, 2023年9月.
4.
竹内 麻由美, 松崎 健司, 坂東 良美, 西村 正人, 原田 雅史 : 子宮アデノマトイド腫瘍のMRI所見の検討, JSAWI (The Japanese Society for the Advancement of Women's Imaging) 第22回シンポジウム, 2021年9月.
竹内 麻由美, 松崎 健司, 西村 正人, 原田 雅史, 苛原 稔 : 付属器腫瘤の鑑別における''preserved follicle sign''の臨床的有用性について, JSAWI (The Japanese Society for the Advancement of Women's Imaging) 第15回シンポジウム, 2017年9月.