Masayuki Takahashi, X J Yang, S McWhinney, Nobuya Sano, C Eng, Susumu Kagawa, B T Teh and Hiro-omi Kanayama : cDNA microarray analysis assists in diagnosis of malignant intrarenal pheochromocytoma originally masquerading as a renal cell carcinoma, Journal of Medical Genetics, Vol.42, 48, 2005.
(要約)
Intrarenal pheochromocytoma (paraganglioma) is a very rare tumour. Its diagnosis is often difficult to establish because of its rarity and its histological similarity to renal cell carcinoma (RCC). Recently, we examined the molecular signatures of different subtypes of kidney tumours by using cDNA microarray. The signature pattern for one tumour, which was originally diagnosed as granular cell RCC, was clearly distinct from that of any other subtype of kidney tumour, and led us to re-evaluate the case. Haematoxylin and eosin staining revealed histological features suggestive of pheochromocytoma, and immunohistochemical studies showed positive staining for neuroendocrine markers but not for keratin. A germline missense mutation, D119E, in the familial paraganglioma related gene succinate dehydrogenase subunit D (SDHD), was subsequently identified. The treatment modality was revised and radiotherapy was given, to which the patient responded, leading to a reduction in tumour size of 25% within the first month. To our knowledge, this is the first report of an intrarenal pheochromocytoma that was diagnosed with the assistance of cDNA microarray analysis.
Masayuki Takahashi, VERONICA PAPAVERO, JASON YUHAS, ERIC KORT, Hiro-omi Kanayama, Susumu Kagawa, ROBERT C. BAXTER, XIMING J. YANG, STEVEN G. GRAY and BIN TEAN TEH : Altered expression of members of the IGF-axis in clear cell renal cell carcinoma, International Journal of Oncology, Vol.26, No.4, 923-931, 2005.
(要約)
Renal cell carcinoma (RCC) is a heterogeneous disease and its biology is poorly understood. The commonest subtype identified is clear cell RCC. The insulin-like growth factor axis is intimately involved with many cellular roles including that of renal development. Dysregulation of this axis has frequently been demonstrated in cancer. In this study, we examine the expression of several IGF-axis components, including receptors, ligands, and binding proteins in clear cell renal cell carcinoma. A series of clear cell RCCs with matched normal kidney from the same individuals were obtained. Total RNA was extracted and expression levels of genes examined using RNase protection analysis. We confirm the dysregulation of the IGF-axis within clear cell renal cell carcinoma including the upregulation of IGFBP-3, which is further validated by immunohistochemical staining on a tissue array containing 50 RCC: positive staining in 29/30 clear cell; 1/10 papillary and 0/10 chromophobe. In addition, we demonstrate that the expression of the A isoform of the insulin receptor is significantly upregulated, while that of IGFBP-5 are significantly downregulated in this tumour subtype.
Tomoharu Fukumori, Natsuo Oka, Toshinori Kasai, Tatsunori Tyubachi, Masayuki Takahashi, Masa-aki Nishitani, Susumu Kagawa and Hiro-omi Kanayama : Expression of galectin-3 is Associated With Resistance to Anticancer Drug-Induced Apoptosis in prostate Cancer, The Journal of Urology, Vol.173, No.4, 60, 2005.
4.
Natsuo Oka, Yasuyo Yamamoto, Masayuki Takahashi, Masaaki Nishitani, Hiro-omi Kanayama and Susumu Kagawa : Expression of angiopoietin-1 and -2, and its clinical significance in human bladder cancer, BJU International, Vol.95, No.4, 660-663, 2005.
(要約)
To investigate the relationship between angiopoietin-1 and -2 expression and the clinicopathological variables and clinical outcome in patients with bladder cancer treated by surgical resection, as both have been recently identified as antagonistic angiogenic factors which regulate tumour growth. The expression of angiopoietin-1 and -2 were assessed by immunohistochemistry in tissue sections from 52 transitional cell carcinomas of the bladder (33 grade 1, 15 grade 2, four grade 3, including two associated with carcinoma in situ; 22 were stage Ta, 19 T1 and 11 T2 tumours). Normal bladder specimens were also resected during each operation as controls. The expression angiopoietins were related to the clinicopathological variables of the tumours. Positive immunostaining was detected in 18 samples (35%) for angiopoietin-1 and in 23 (44) for angiopoietin-2. There was no significant difference in survival according to tumour angiopoietin-1 status in the patients, but in contrast the overall survival of patients with angiopoietin-2-positive tumours was significantly lower than for those with angiopoietin-2-negative tumours (P < 0.05). Positive angiopoietin-2 expression was significantly correlated with histological grade (P = 0.026), histological stage (P = 0.009) and poor prognosis (P < 0.05). On multivariate analysis, positive angiopoietin-2 expression was an independent negative predictor for survival (P = 0.042). These results suggest that angiopoietin-2 overexpression is associated with tumour progression, thereby indicating a poor prognosis for some patients treated by surgical resection for bladder carcinoma.
Masumi Okamoto, Koichiro Tsuchiya, Yasuhisa Kanematsu, Yuki Izawa, Masanori Yoshizumi, Susumu Kagawa and Toshiaki Tamaki : Nitrite-derived nitric oxide formation following ischemia-reperfusion injyury in kidney., American Journal of Physiology, Renal Physiology, Vol.288, No.1, 182-187, 2005.
(要約)
Nitric oxide (NO) is synthesized from l-arginine by nitric oxide synthase (NOS), and nitrite and nitrate are believed to be waste forms of NO. We previously reported an enzyme-independent pathway of NO generation from nitrite in acidic conditions. In this study, we show nitrite-derived NO formation in renal ischemia-reperfusion injury using electron paramagnetic resonance (EPR) spectroscopy. In this experiment, we utilized a stable isotope of [(15)N]nitrite as a source of nitrite to distinguish l-arginine-derived NO from [(15)N]nitrite-derived (15)NO. Intravenous infusion of a stable isotope of [(15)N]nitrite ((15)NO(2)(-)) facilitated the formation of Hb(15)NO during renal ischemia, which demonstrated that the origin of NO was nitrite. The EPR signal of Hb(15)NO in kidney appeared after 40 min of renal ischemia, and renal reperfusion decreased the Hb(15)NO level in the kidney and increased it in blood by contrast. In addition, the amount of HbNO was nitrite concentration dependent, and this formation was NOS independent. Our findings suggest that nitrite can be an alternative source of NO in ischemic kidney and that it binds with hemoglobin and then is spread by the circulation after reperfusion.
Kyle A. Furge, Kerry A. Lucas, Masayuki Takahashi, Jun Sugimura, Eric J. Kort, Hiro-omi Kanayama, Susumu Kagawa, Philip Hoekstra, John Curry, Ximing J. Yang and Bin T Teh : Robust Classification of Renal Cell Carcinoma Based on Gene Expression Data and Predicted Cytogenetic Profiles, Cancer Research, Vol.64, No.12, 4117-4121, 2004.
(要約)
Renal cell carcinoma (RCC) is a heterogeneous disease that includes several histologically distinct subtypes. The most common RCC subtypes are clear cell, papillary, and chromophobe, and recent gene expression profiling studies suggest that classification of RCC based on transcriptional signatures could be beneficial. Traditionally, however, patterns of chromosomal alterations have been used to assist in the molecular classification of RCC. The purpose of this study was to determine whether it was possible to develop a classification model for the three major RCC subtypes that utilizes gene expression profiles as the bases for both molecular genetic and cytogenetic classification. Gene expression profiles were first used to build an expression-based RCC classifier. The RCC gene expression profiles were then examined for the presence of regional gene expression biases. Regional expression biases are genetic intervals that contain a disproportionate number of genes that are coordinately up- or down-regulated. The presence of a regional gene expression bias often indicates the presence of a chromosomal abnormality. In this study, we demonstrate an expression-based classifier can distinguish between the three most common RCC subtypes in 99% of cases (n = 73). We also demonstrate that detection of regional expression biases accurately identifies cytogenetic features common to RCC. Additionally, the in silico-derived cytogenetic profiles could be used to classify 81% of cases. Taken together, these data demonstrate that it is possible to construct a robust classification model for RCC using both transcriptional and cytogenetic features derived from a gene expression profile.
H Hayashi, M Furihata, M Kuwahara, Susumu Kagawa, T Shuin and Y Ohtsuki : Infrequent Alteration in the p53R2 Gene in Human Transitional Cell Carcinoma of the Urinary Tract, Pathobiology, Vol.71, No.2, 103-106, 2004.
(要約)
Functional defects in DNA repair have been shown to be associated with genomic instability followed by cancer. Recently, p53R2 [p53-inducible ribonucleotide reductase (RR) small subunit 2 homologous] was identified as a novel RR gene which is directly regulated by p53 protein in the G1 and G2 phases of the cell cycle supplying nucleotides to repair damaged DNA. We performed genetic analyses of p53R2 to determine whether p53R2 alterations play significant roles in urothelial tumorigenesis. Genomic DNA from 108 primary transitional cell carcinomas (TCCs; 81 of the urinary bladder and 27 of the renal pelvis or ureter) was analyzed for mutation in the p53R2 gene by direct sequencing. We focused on three domains of the p53R2 gene: one RR small subunit signature involving codons 120-146 (region 1) and two putative nuclear localization signal sequences, involving codons 149-155 (region 2) and codons 163-169 (region 3). In addition, a p53-binding site of 20 nucleotides in intron 1 of p53R2 was also analyzed. One renal pelvic TCC (0.9%: 1/108) had a single-base substitution in p53R2 with a G to T transversion resulting in the amino acid substitution Glu136 --> Asp. This base substitution was localized within the domain of exon 4 encoding the RR small subunit signature, and causes an amino acid substitution in one of the most highly conserved regions of p53R2, in which human R2 and yeast RNR2 and RNR4 proteins are highly homologous. This finding provides the in vivo evidence for the infrequent involvement of alterations in p53R2 inhuman urothelial TCCs.
(キーワード)
Aged / Aged, 80 and over / Amino Acid Substitution / Base Sequence / Carcinoma, Transitional Cell / Cell Cycle Proteins / DNA, Neoplasm / Female / Humans / Male / Middle Aged / Molecular Sequence Data / Neoplasm Staging / Polymerase Chain Reaction / Ribonucleotide Reductases / Urologic Neoplasms
Natsuo Oka, Yuushi Okumura, Hiro-omi Kanayama, Hirofumi Izaki, Masumi Okamoto, Hiroshi Kido and Susumu Kagawa : Amiloride and Urinary Trypsin Inhibitor Inhibit Urothelial Cancer Invasion, European Urology, Vol.44, No.6, 737-741, 2003.
(要約)
To determine the contribution of urokinase-type plasminogen activator (uPA) and plasmin in the invasion of highly invasive urothelial cancer cells. We compared expression levels of mRNA and protease activity of uPA and plasmin formation in primary cultures of the noninvasive transitional cell carcinoma, UCT-1, and in the highly invasive type, UCT-2. By using in vitro cell invasion assay system, we evaluated the effects of amiloride and urinary trypsin inhibitor (UTI), which inhibit uPA and plasmin, respectively, on invasion by both cell lines. Expression levels of mRNA, protein, and activities of uPA were significantly higher (p<0.005) and resulted in more plasminogen activation in UCT-2 than in UCT-1. Amiloride and UTI significantly inhibited plasmin formation and the invasion of both cell lines (p<0.001). High expression levels of mRNA, activities of uPA and high plasmin formation significantly potentiated the invasiveness of urothelial cancer cells. Thus, inhibitors of uPA and plasmin, such as amiloride and UTI, respectively, could be useful therapeutic tools with which to treat urothelial cancer.
Tomoharu Fukumori, Yukinori Takenaka, Tadashi Yoshii, Hyeong-Reh Choi Kim, Victor Hogan, Hidenori Inohara, Susumu Kagawa and Avraham Raz : CD7 and CD29 mediate galectin-3 induced T cell apoptosis., Cancer Research, Vol.63, No.23, 8302-8311, 2003.
(要約)
Galectin (Gal)-3, a M(r) 31000 member of the beta-galactoside-binding protein family, is a multifunctional protein implicated in a variety of biological functions, including tumor cell adhesion, proliferation, differentiation, angiogenesis, apoptosis, cancer progression, and metastasis. Here, we report that secreted extracellular Gal-3 can signal apoptosis of human T leukemia cell lines, human peripheral blood mononuclear cells, and activated mouse T cells after binding to cell surface glycoconjugate receptors through carbohydrate-dependent interactions because the apoptotic effect was found to be inhibited by lactose, specific sugar inhibitor, and to be dose dependent. However, the apoptosis sensitivity to Gal-3 varied among the different cell lines tested. We report that Gal-3-null Jurkat, CEM, and MOLT-4 cells were significantly more sensitive to exogenous Gal-3 than SKW6.4 and H9 cells, which express Gal-3, suggesting a cross-talk between the antiapoptotic activity of intracellular Gal-3 and proapoptotic activity of extracellular Gal-3. Furthermore, Gal-3-transfected CEM cells were found to be more resistant to C(2)-ceramide-induced apoptosis than the control CEM cells. Identification of Gal-3 cell surface receptors revealed that Gal-3 binding to CD7 and CD29 (beta(1) integrin) induced apoptosis. Gal-3 binding to its cell surface receptors results in activation of mitochondrial apoptosis events including cytochrome c release and caspase-3 activation, but not caspase-8 activation. Taken together, these results suggest that the induction of T-cell apoptosis by secreted Gal-3 may play a role in the immune escape mechanism during tumor progression through the induction of apoptosis to cancer-infiltrating T cells. The induction of T-cell apoptosis by secreted Gal-3 is dependent in part on the presence or absence of cytoplasmic Gal-3, providing a new insight for the immune escape mechanism of cancer cells.
Jindong Chen, Weng-Onn Lui, Michele D. Vos, Geoffrey J. Clark, Masayuki Takahashi, Jacqueline Schoumans, Sok Kean Khoo, David Petillo, Todd Lavery, Jun Sugimura, Dewi Astuti, Chun Zhang, Susumu Kagawa, Eamonn R. Maher, Catharina Larsson, Arthur S. Alberts, Hiro-omi Kanayama and Bin Tean Teh : The t(1;3) breakpoint-spanning genes LSAMP and NORE1 are involved in clear cell renal cell carcinomas, Cancer Cell, Vol.4, No.5, 405-413, 2003.
(要約)
By positional cloning, we identified two breakpoint-spanning genes in a familial clear cell renal cell carcinoma (CCRCC)-associated t(1;3)(q32.1;q13.3): LSAMP and NORE1 (RASSF1 homolog). Both genes are downregulated in 9 of 9 RCC cell lines. While the NORE1A promoter predominantly presents partial methylation in 6 of the cell lines and 17/53 (32%) primary tumors, the LSAMP promoter is completely methylated in 5 of 9 cell lines and in 14/53 (26%) sporadic and 4 familial CCRCCs. Expression of LSAMP and NORE1A proteins in CCRCC cell lines inhibited cell proliferation. These characteristics indicate that LSAMP and NORE1A may represent new candidate tumor suppressors for CCRCC.
Yasuyo Yamamoto, Noriyuki Nishimura, Shinya Morimoto, Hiroko Kitamura, Shinji Manabe, Hiro-omi Kanayama, Susumu Kagawa and Takuya Sasaki : Distinct roles of Rab3B and Rab13 in the polarized transport of apical, basolateral, and tight junctional membrane proteins to the plasma membrane, Biochemical and Biophysical Research Communications, Vol.308, No.2, 270-275, 2003.
(要約)
Regulated transport of proteins to distinct plasma membrane domains is essential for the establishment and maintenance of cell polarity in all eukaryotic cells. The Rab family small G proteins play a crucial role in determining the specificity of vesicular transport pathways. Rab3B and Rab13 localize to tight junction in polarized epithelial cells and cytoplasmic vesicular structures in non-polarized fibroblasts, but their functions are poorly understood. Here we examined their roles in regulating the cell-surface transport of apical p75 neurotrophin receptor (p75NTR), basolateral low-density lipoprotein receptor (LDLR), and tight junctional Claudin-1 using transport assay in non-polarized fibroblasts. Overexpression of Rab3B mutants inhibited the cell-surface transport of LDLR, but not p75NTR and Claudin-1. In contrast, overexpression of Rab13 mutants impaired the transport of Claudin-1, but not LDLR and p75NTR. These results suggest that Rab3B and Rab13 direct the cell-surface transport of LDLR and Claudin-1, respectively, and may contribute to epithelial polarization.
AHMED ANWAR, YASUSHI KUROKAWA, Masayuki Takahashi, YASUYO YAMAMOTO, Hiro-omi Kanayama and Susumu Kagawa : Functional evaluation of one-stage urethroplasty with parameatal foreskin flaps repair of hypospadias using uroflowmetry, International Journal of Urology, Vol.10, No.6, 297-301, 2003.
(要約)
Uroflowmetry is a simple, accurate and non-invasive test. In the present study, we aimed to determine the role of uroflowmetry in the evaluation of the functional results of one-stage urethroplasty with parameatal foreskin flaps (OUPF) technique. Twenty-one children who had undergone OUPF repair at our clinic were selected. Selection criteria were that patients were toilet trained and had no fistula. Uroflowmetry was performed using a rotating disk sensor. The maximum flow rate (Qmax) and average flow rate (Qave) were plotted against body surface related flow rate nomograms. The upper 95% tolerance limits for the 5th, 10th, 15th, 20th and 25th percentiles of the normal population were used for comparison. The flow pattern was classified as bell ring, plateau or intermittent. The median age at the first uroflowmetry was 4.7 years (range 2.5-8.6) and the mean postoperative follow-up period was 25 months (range 1-58). Twelve children had Qmax above the 25th, six between the 5th and 25th and three less than the 5th percentiles of the normal population. A normal bell-shaped flow curve was obtained in 17 (80.6%) of the children. Of the three children with Qmax below the 5th percentile, two children had a plateau flow pattern and were found to have a urethal stricture. Dilation was performed successfully, after which the Qmax returned to the normal range and the symptoms disappeared. The OUPF technique provided satisfactory functional results for hypospadias repair. We advocate the use of uroflowmetry for routine postoperative follow-up.
Yasushi Kurokawa, Hiro-omi Kanayama, Ahmed Anwar, Tomoharu Fukumori, Yasuyo Yamamoto, Masayuki Takahashi, Susumu Kagawa, Yosihide Murakami and Toshiro Terachi : Laparoscopic nephroureterectomy for dysplastic kidney in children: an initial experience, International Journal of Urology, Vol.9, No.11, 613-617, 2002.
(要約)
Laparoscopic nephroureterectomy for dysplastic kidney is now becoming a widely accepted procedure. We report here our initial experience with laparoscopic nephroureterectomy in four girls. Between 1993 and 1999, laparoscopic nephroureterectomy was performed in four girls (mean age 5.3 years). Three patients had an ectopic dysplastic kidney with ectopic ureter, and one patient had hydronephrosis with megaureter due to distal ureteral atresia of the upper moiety in a duplicated dysplastic kidney. The transperitoneal approach was used in all cases. Mean operative time was 195 min (range 150-266). Blood loss was minimal. All operations were completed successfully and there were no intraoperative or postoperative complications except for subcutaneous emphysema in one patient (case 4). Postoperative analgesia was used in three patients and administered in the form of diclofenac sodium suppositories 12.5 mg (cases 1 and 2) or acetaminophen suppositories 50 mg (case 3) for 1-2 days. One patient did not require any analgesia (case 4). Oral fluid intake was resumed on the first postoperative day and ambulation began within 1-3 days (mean 1.6, cases 1, 2 and 3) and 6 days (case 4). All children returned to normal activity within 3-6 days of surgery. Mean postoperative hospital stay was 7.3 days. All cases had uneventful courses after discharge. Laparoscopic nephroureterectomy can be performed safely, with minimal postoperative pain, excellent cosmetic results and early ambulation. We advocate the use of laparoscopy for the diagnosis and treatment of dysplastic kidney with ectopic ureter.
(キーワード)
children / dysplastic kidney / laparoscopic nephroureterectomy
Tomoharu Fukumori, Masaaki Nishitani, Takushi Naroda, Tomoichiro Onishi, Natsuo Oka, Hiro-Omi Kanayama and Susumu Kagawa : Expression of angiostatin cDNA in a murine renal cell carcinoma suppresses tumor growth in vivo., Urology, Vol.59, No.6, 973-977, 2002.
(要約)
To investigate the effectiveness of angiostatin gene therapy for renal cancer using a mouse model. The generally poor prognosis of advanced renal cancer indicates the need for new therapeutic modalities. The dependency of solid tumor growth on angiogenesis suggests that antiangiogenic therapy would be effective against renal cell carcinoma, which is generally a hypervascular tumor. Murine renal cancer cells (Renca) transfected with murine angiostatin cDNA (AST-Renca) were subcutaneously implanted in BALB/c mice. Subsequently, the macroscopic appearance and volume of tumors were evaluated once per week. Renca cells transfected with empty plasmid DNA (mock-Renca) were used as a control. In addition, histologic sections of tumor were analyzed for neovascularization on the basis of an immunohistochemical analysis for CD31. The antitumor effect of AST-Renca on a parental Renca tumor at a distant site was also evaluated. The mean volume of AST-Renca tumors was significantly less than that of the control vector-transfected tumors 3 weeks after implantation. In the cell proliferation assay, the expression of angiostatin did not inhibit the proliferation of Renca cells in vitro. Immunohistochemical analysis of neovascularization by staining with anti-CD31 antibody revealed that angiostatin suppressed tumor vessel formation. Moreover, implantation of AST-Renca inhibited the growth of parental Renca implanted simultaneously at a distant site. Expression of an angiostatin transgene can suppress the growth of murine renal cancer through the inhibition of tumor-induced angiogenesis. Angiostatin gene therapy may be effective against renal cancer.
Masayuki Takahashi, Daniel R. Rhodes, Kyle A. Furge, Hiro-omi Kanayama, Susumu Kagawa, Brian B. Haab and Bin Tean Teh : Gene expression profiling of clear cell renal cell carcinoma: Gene identification and prognostic classification, Proceedings of the National Academy of Sciences of the United States of America, Vol.98, No.17, 9754-9759, 2001.
(要約)
To better understand the molecular mechanisms that underlie the tumorigenesis and progression of clear cell renal cell carcinoma (ccRCC), we studied the gene expression profiles of 29 ccRCC tumors obtained from patients with diverse clinical outcomes by using 21,632 cDNA microarrays. We identified gene expression alterations that were both common to most of the ccRCC studied and unique to clinical subsets. There was a significant distinction in gene expression profile between patients with a relatively nonaggressive form of the disease [100% survival after 5 years with the majority (15/17 or 88%) having no clinical evidence of metastasis] versus patients with a relatively aggressive form of the disease (average survival time 25.4 months with a 0% 5-year survival rate). Approximately 40 genes most accurately make this distinction, some of which have previously been implicated in tumorigenesis and metastasis. To test the robustness and potential clinical usefulness of this molecular distinction, we simulated its use as a prognostic tool in the clinical setting. In 96% of the ccRCC cases tested, the prediction was compatible with the clinical outcome, exceeding the accuracy of prediction by staging. These results suggest that two molecularly distinct forms of ccRCC exist and that the integration of expression profile data with clinical parameters could serve to enhance the diagnosis and prognosis of ccRCC. Moreover, the identified genes provide insight into the molecular mechanisms of aggressive ccRCC and suggest intervention strategies.
Natsuo Oka, Tomoharu Fukumori, Masayuki Takahashi, Hiro-omi Kanayama and Susumu Kagawa : Secondary amyloidosis of the bladder causing macroscopic hematuria, International Journal of Urology, Vol.8, No.6, 330-332, 2001.
(要約)
Bladder involvement in amyloidosis is unusual. The case of an 80-year-old man with macroscopic hematuria caused by secondary amyloidosis of the bladder is described. Cystoscopic examination revealed only a diffuse edematous area and bleeding. No tumor-like lesions were identified. Transurethral biopsy revealed amyloid deposits. Macroscopic hematuria disappeared spontaneously after cystoscopy and bladder biopsy. The patient has been followed up without treatment and is currently free of symptoms.
(キーワード)
Aged / Aged, 80 and over / Amyloidosis / Hematuria / Humans / Male / Urinary Bladder
Hiro-omi Kanayama, Weng-Onn Lui, Masayuki Takahashi, Takushi Naroda, Darek Kedra, Fung Ki Wong, Yoko Kuroki, Yutaka Nakahori, Catharina Larsson, Susumu Kagawa and Bin Tean Teh : Association of a novel constitutional translocation t(1q;3q) with familial renal cell carcinoma, Journal of Medical Genetics, Vol.38, No.3, 165-170, 2001.
(要約)
Four cases of late onset clear cell renal cell carcinoma (RCC), a case of gastric cancer, and a case of exocrine pancreatic cancer were identified in a Japanese family. In order to elucidate the underlying mechanism for tumorigenesis in this family, extensive genetic studies were performed including routine and spectral karyotyping (SKY), fluorescence in situ hybridisation (FISH), comparative genomic hybridisation (CGH), loss of heterozygosity studies (LOH), and VHL mutation analysis. A germline translocation t(1;3)(q32-q41;q13-q21) was identified by karyotyping in five members of the family including all three RCC cases tested. The translocation was refined to t(1;3)(q32;q13.3) by FISH analysis using locus specific genomic clones, and the two breakpoints were mapped to a 5 cM region in 3q13.3 and a 3.6 cM region in 1q32. Both CGH and allelotyping using microsatellite markers showed loss of the derivative chromosome 3 carrying a 1q segment in the three familial RCCs analysed. Additional chromosomal imbalances were identified by CGH, including amplifications of chromosomes 5 and 7 and loss of 8p and 9. No germline VHL mutation was found but two different somatic mutations, a splice (IVS1-2A>C) and a frameshift (726delG), were identified in two RCCs from the same patient confirming their distinct origin. Taken together, these results firmly support a three step model for tumorigenesis in this family. A constitutional translocation t(1q;3q) increased the susceptibility to loss of the derivative chromosome 3 which is then followed by somatic mutations of the RCC related tumour suppressor gene VHL located in the remaining copy of chromosome 3.
(キーワード)
familial renal cell carcinoma / translocation / von Hippel-Lindau disease / loss of heterozygosity
K Kanda, Masayuki Takahashi, Yoshihide Murakami, Hiro-omi Kanayama and Susumu Kagawa : The role of the activated form of matrix metalloproteinase-2 in urothelial cancer, BJU International, Vol.86, No.4, 553-557, 2000.
(要約)
To investigate the expression of matrix metalloproteinase-2 (MMP-2, reportedly associated with cancer cell invasion and metastasis in many human cancers) in urothelial tumours and thus define its role in this disease. Materials and methods The expression of both the activated form of MMP-2 and total MMP-2 (activated + latent form) was measured using gelatine zymography in tissue obtained surgically from 61 patients with urothelial cancer. The correlation between the level of the activated form of MMP-2 and clinical and histological variables was assessed. The expression of activated and total MMP-2 were significantly higher in invasive tumour tissue and both levels were correlated with histological grade. In particular, the level of activated MMP-2 was more closely correlated than that of total MMP-2 in invasive tumour tissue. Moreover, high levels of activated MMP-2 were strongly associated with shorter disease-specific survival (P = 0.0016). These findings suggest that activated MMP-2 plays a significant role in invasion of urothelial cancer and that the level of activated MMP-2 expression is a useful prognostic indicator.
Tomoharu Fukumori, Akari Hirofumi, Yoshida Akiko, Mikako Fujita, Koyama AH, Susumu Kagawa and Akio Adachi : Regulation of cell cycle and apoptosis by human immunodeficiency virus type 1 Vpr., Microbes and Infection, Vol.2, No.9, 1011-1017, 2000.
(要約)
Biological effects of HIV-1 Vpr on CD4(+) cells were studied by an infection system. High-titered HIV-1 stocks pseudotyped with vesicular stomatitis virus G protein were prepared and used to inoculate into CD4(+ )T cells at high multiplicity of infection. Both cell- and virion-associated Vpr were demonstrated to arrest the cell cycle at the G2/M phase, and to induce cell apoptosis. Of note, morphologically apoptotic cells were shown to be arrested at the G2/M stage. No appreciable effect of Vpr on the anti-Fas antibody-mediated apoptosis was observed in this system.
Li NC, Kazaya Kanda, Tomoharu Fukumori, Yoshio Inoue, Masa-aki Nishitani, Hiro-omi Kanayama and Susumu Kagawa : The role of expression of vascular endothelial growth factor isoforms and platelet-derived endothelial cell growth factor in bladder cancer., Urologic Oncology, Vol.5, 1-6, 2000.
26.
Tomoharu Fukumori, Susumu Kagawa, Shinya Iida, Yoko Oshima, Hirofumi Akari, Hajime A Koyama and Akio Adachi : Rev-dependent expression of three species of HIV-1 mRNAs, International Journal of Molecular Medicine, Vol.3, No.3, 297-302, 1999.
(要約)
The expression of structural and accessory genes of human immunodeficiency virus type 1 (HIV-1) except for nef requires a viral regulatory protein Rev. Rev-dependency of the expression of structural (gag, pol and env), regulatory (tat and rev), and accessory genes (vif, vpr, vpu and nef) has been investigated by various systems, and it has been demonstrated that unspliced (encodes gag and pol) and singly-spliced (env-vpu, vif and vpr) viral mRNAs are differentially dependent on the function of Rev. In this review, the function of HIV-1 Rev in relation to these findings is discussed.
Tomoharu Fukumori, Hirofumi Akari, Shinya Iida, So Hata, Susumu Kagawa, Yoko Aida, Hajime A Koyama and Akio Adachi : The HIV-1 Vpr displays strong anti-apoptotic activity, FEBS Letters, Vol.432, No.1-2, 17-20, 1998.
(要約)
Mutations in the human immunodeficiency virus type 1 (HIV-1) vpr gene only slightly reduce the replication rate of the virus. To study the role of HIV-1 Vpr in biological effects on cells, HEp-2 cells, which express HIV-1 Vpr constitutively but at a low level, were established. While control HEp-2 cells underwent apoptosis when incubated with sorbitol, the morphological and biochemical apoptotic changes were inefficiently induced in the HIV-1 Vpr-expressing cells by the same treatment. These results clearly indicate that HIV-1 Vpr has anti-apoptotic activity, and raise the possibility that Vpr acts as a weak activator of virus replication through anti-apoptosis.
Atsuko Furukawa, Kenichi Maeda, Masayuki Miyasaka, Susumu Kagawa, Koji Yasutomo, Hajime Hisaeda, HIdeyuki Nagasawa and Kunisuke Himeno : Establishment of a xenogeneic chimera without GVHD in NK cell-depleted SCID mice by grafting rat fetal liver cells, Cellular Immunology, Vol.164, No.2, 176-181, 1995.
(要約)
Rat lymphopoiesis did not develop when naive SCID mice were transplanted with rat fetal liver cells. However, when SCID mice were depleted of NK cells by administration of anti-murine IL-2R beta mAb before transplantation, remarkable reconstitution of both rat T and B cells was observed in these mice without any evidence of graft-versus-host disease macroscopically or histologically. T cells in these reconstituted mice proliferated well in response to Con A and third-party rat and mouse antigens, whereas no response was seen to the stimulation with either donor rat- or host mouse-type cells. When these xenogeneic chimera mice had been immunized with SRBC, these mice exhibited DTH reaction and antibody production against the homologous antigen. These results indicate that rat fetal liver cells can differentiate to functional T and B cells in the xenogeneic microenvironment of SCID mice, if host NK cells are depleted beforehand. These rat-type T cells develop within SCID thymuses and acquire tolerance to either donor F344 rat or host SCID mouse antigens.
Masaharu Kan, Fumihiko Kanai, Mitsuru Iida, Hideaki Jinnouchi, Mikio Todaka, Takanobu Imanaka, Kimio Ito, Yasuhiko Nishioka, Tetsuo Ohnishi, Seika Kamohara, Hideaki Hayashi, Takashi Murakami, Susumu Kagawa, Hiroyuki Sano, Naotake Hashimoto, Sho Yoshida, Hideichi Makino and Yousuke Ebina : Frequency of Mutations of Insulin Receptor Gene in Japanese Patients with NIDDM, Diabetes, Vol.44, No.9, 1081-1086, 1995.
(要約)
To examine the prevalence of abnormalities in the insulin receptor structure gene in Japanese with non-insulin-dependent diabetes mellitus (NIDDM), a population of 51 patients with NIDDM was screened for mutations in this gene. Patient genomic DNAs of both alleles corresponding to 22 exons of the gene were amplified by polymerase chain reaction (PCR). The PCR products on pUC19 were sequenced. Three patients with heterozygous missense mutation Thr831-->Ala831 in exon 13 and one patient with heterozygous missense mutation Tyr1334-->Cys1334 in exon 22 of the beta-subunits were identified. Linkage analysis of one of the families plus statistical studies showed that the mutation Thr831-->Ala831 is possibly responsible for the onset of NIDDM. In COS cells transiently expressing both mutant receptor cDNAs and a cDNA of a M(r) 85,000 regulatory subunit of phosphatidylinositol 3-kinase (PI 3-kinase), the mutation Tyr1334-->Cys1334 impaired binding of the receptor with the M(r) 85,000 subunit of PI 3-kinase, but linkage analysis of the family showed that the mutation did not cosegregate with NIDDM in the pedigree. Therefore, one missense mutation (Thr831-->Ala831) in the insulin receptor, as found in three patients, is possibly involved in the etiology of a subset of the 51 NIDDM patients.
(キーワード)
Adult / Aged / Alanine / Amino Acid Sequence / Animals / Base Sequence / Cell Line / Cercopithecus aethiops / Cysteine / DNA / DNA Primers / Diabetes Mellitus, Type 2 / Exons / Female / Genetic Linkage / Humans / Insulin / Japan / Kinetics / Macromolecular Substances / Male / Middle Aged / Molecular Sequence Data / Pedigree / Phosphatidylinositol 3-Kinases / Phosphotransferases (Alcohol Group Acceptor) / Point Mutation / Polymerase Chain Reaction / Receptor, Insulin / Recombinant Proteins / Threonine / Transfection / Tyrosine
Hiroyuki Inoue, Hitoshi Miki, Kazushi Oshimo, Katuhiro Tanaka, Yasumasa Monden, Akihiro Yamamoto, Susumu Kagawa, Nobuya Sano, Eiji Hayashi, Masaru Nagayama and Yoshio Hayashi : Familial hyperparathyroidism associated with jaw fibroma: case report and literature review, Clinical Endocrinology, Vol.43, No.2, 225-229, 1995.
(要約)
A 53-year-old female suffering from renal stones and hypercalcaemia was diagnosed as having primary hyperparathyroidism caused by hyperplasia of the parathyroid glands. She underwent total parathyroidectomy and implantation of parathyroid tissue. After one year, she underwent surgery for a jaw tumour. The pathological findings indicated it to be a cementifying fibroma. Jackson et al. (1990) reported the familial association of hyperparathyroidism with jaw tumours, and they suggested that this condition represents a new clinical syndrome. We believe that our case belongs to this syndrome.
Mowla Mohammed Golam Chowdhury, Atsuko Furukawa, Susumu Kagawa, Kenichi Maeda, Koji Yasutomo and Kunisuke Himeno : B cells are required as APC for antigen-specific T cell proliferation but not the differentiation or priming of those T cells, The Tokushima Journal of Experimental Medicine, Vol.41, No.1-2, 1-8, 1994.
(要約)
We studied the influences of B cells on functional differentiation of T cells using SCID mice grafted with fetal thymus of C.B-17 mice (TG mice). T cells were shown to be reconstituted in TG mice without B cell development. These mice showed normal DTH response to SRBC and OVA. LN cells of these mice produced cytokines including IL-2, IL-4, IL-6 and IFN-gamma according to Con A stimulation. Thus, majority of T cell functions seem to differentiate in the absence of B cells. However, T cells of TG mice failed to proliferate in response to immunizing antigens in vitro, although they responded well to stimulation with Con A. This unresponsiveness of T cells in TG mice to these antigens was restored when antigen-primed B cells were added to the proliferation assay. Such an inability of T cells in antigen-specific proliferation was not seen in SCID mice grafted with C.B-17 fetal liver cells, in which B cells as well as T cells were efficiently reconstituted (FLT mice). T cell proliferation to immunizing antigen was also abrogated in FLT mice when B cells were depleted from lymphoid population. These results indicate that T cells can functionally differentiate and be primed in the absence of B cells, but they require B cells to proliferate in response to foreign antigens.
Yoshihiro Tsuruo, Kazunori Ishimura, Masato Tamura, Susumu Kagawa and Kyoji Morita : Biochemical and histochemical studies of the effects of cerebral metabolism-improving drugs on NADPH diaphorase in the mouse brain, The Japanese Journal of Pharmacology, Vol.65, No.3, 285-288, 1994.
(要約)
The effects of cerebral metabolism-improving drugs on NADPH diaphorase activity in the mouse brain were studied, and we found that diaphorase activity in the post-mitochondrial fraction of brain homogenate was enhanced by idebenone in a concentration-dependent manner. Histochemical studies also indicated that diaphorase staining was intensified by idebenone at the same concentration. These results suggest that idebenone may stimulate the production of nitric oxide, probably through its direct action on nitric oxide synthase, thus producing its protective action on neurological disorders due to cerebral hypoxia or ischemia as a consequence of dilating the cerebral blood vessels.
Masato Tamura, Susumu Kagawa, Yoshihiro Tsuruo, Kazunori Ishimura and Kyoji Morita : Effects of flavonoid compounds on the activity of NADPH diaphorase prepared from the mouse brain, The Japanese Journal of Pharmacology, Vol.65, No.4, 371-373, 1994.
(要約)
The effects of flavonoids on NADPH diaphorase activity were studied in vitro, and we found that the enzyme activity was markedly inhibited by quercetin. This inhibitory action was shown to be accompanied by an increase in the apparent Km value of the enzyme for the cofactor NADPH, with a decrease in the Vmax, and an increase in the apparent Km for the substrate nitro blue tetrazolium, without any significant change in the Vmax. These results indicate that quercetin may directly inhibit NADPH diaphorase, thus suggesting the possibility that this compound may be able to inhibit the production of nitric oxide in the brain.
Chowdhury Mowla Golam, Akihiro Yamamoto, Hiroomi Kanayama, Masashi Aki, Susumu Kagawa, Muneo Nakamura and Nobuya Sano : ADENOMATOUS POLYP OF THE PROSTATIC URETHRA: A CASE REPORT, The Nishinihon Journal of Urology, Vol.54, No.10, 1735-1738, 1992.
Tomoharu Fukumori, Masa-aki Nishitani, M Takahashi, T Naroda, Hirofumi Izaki, I Elsamman, Susumu Kagawa and Hiro-omi Kanayama : Involvement of Matrix Metalloproteinase-3 in Hepatocyte Growth Factor-Induced Invasion of Bladder Cancer, BUJ International, Honolulu, Oct. 2004.