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UP-2.065: Expression of GAT1 in Male Reproductive System and Its Effects on Reproduction in Mice
UNMODERATED POSTER SESSIONS
ejaculatory duct and prostate. Among all the cases followed up more than 3 to 51 months after operation, 36 patients (81.8%) had improved semen parameters and 11 patients’ wives (25%) had pregnancies in 44 cases of TURED, 6 patients (60%) had improved semen parameters and 3 patients’ wives (30%) had pregnancies in 10 cases of vasoepididymostomy. Conclusion: Semen analyses of fructose and alpha-glucosidase measurement in seminal plasma, TRUS and vasography are major diagnostic methods for obstructive azoospermia. TURED and vasoepididymostomy may separately be effective methods for the treatment of azoospermia with EDO and obstructive azoospermia with epididymal level.
UP-2.065 Expression of GAT1 in Male Reproductive System and Its Effects on Reproduction in Mice Zhang J, Gui Y, Yuan T, Bian C Department of Urology, Tongji Hospital of Tongji University, Shanghai, China Objective: The present study was carried out to identify GABA transport protein I (GAT1) in male reproductive organs and to study the effect of GAT1 overexpression in the male reproductive system of GAT1 transgenic mice (TG). Materials and Methods: Expression and location of GAT1 in testes, epididymis and sperm of wild-type (WT) mice were identified by immunohistochemistry and westernblot. Histological changes of testis, epididymis and sperm of transgenic mice overexpressing GAT1 were detected by immunofluorescent staining and HE staining. Results: GAT1 expression was detected in the testis, epididymis and sperm of non-transgenic mice. Vacuolization and deformity of spermatogenic cells were observed in the transgenic mice, but the epididymis was unremarkable. Immunofluorescent staining showed that the number of diastrophic and decapitated sperm increased significantly in transgenic mice to 46.9% from 7.3% in nontransgenic mice. Conclusion: These results suggest that abnormal expression of GAT1 could result in spermogenesis function injury, sperm paramorphia and dysgenesis.
UP-2.066 Increased Serum Hepcidin-25 Level and Increased Tumor Expression of Hepcidin MRNA is Associated with Metastasis of Renal Cell Carcinoma
S252
Abe H1, Tomosugi N2, Kamai T1, Arai K1, Yoshida K1 1 Department of Urology, Dokkyo Medical University, Tochigi, Japan; 2Proteomics Research Unit, Division of Advanced Medicine, Medical Research Institute, Kanazawa Medical University, Ishikawa, Japan Introduction and Objective: Hepcidin is a molecule that is important for iron metabolism. We investigated the role of hepcidin in renal cell carcinoma (RCC). Materials and Methods: We measured serum hepcidin-25 levels in 32 patients by liquid chromatograpy (LC)-mass spectrometry (MS)/MS, and assessed hepcidin mRNA expression in paired tumor and non-tumor tissue samples from the surgical specimens of 53 consecutive patients with RCC by using a real-time reverse transcription polymerase chain reaction. Results: The serum hepcidin-25 level was higher in metastatic RCC than in nonmetastatic RCC (P⬍0.0001). Expression of hepcidin mRNA was lower in tumor tissue than in non-tumor tissues (P⬍0.0001). The serum hepcidin-25 level was not correlated with the expression of hepcidin mRNA in the corresponding tumor tissue specimens from 32 patients. Hepcidin mRNA expression in tumor tissue was correlated with metastatic potential, but not with tumor differentiation or the local stage. Kaplan-Meier analysis showed that high hepcidin mRNA expression was related to shorter overall survival in all patients. Univariate analysis according to the Cox proportional hazards model showed that hepcidin mRNA level was an independent prognostic factor in overall survival. Conclusions: Our findings suggest that a high serum hepcidin-25 level may be an indicator of the progression of RCC, and that upregulation of hepcidin mRNA in tumor tissue may be related to increased metastatic potential. UP-2.067 The Target of Cysteinyl-Leukotriene1 Receptor (CysLT1R) Is New AntiCancer Strategy for Human Renal Cell Carcinoma Funao K, Matsuyama M, Kuratsukuri K, Tanaka T, Takemoto Y, Nakatani T, Yoshimura R Dept. of Urology, Osaka City University, Graduate School of Medicine, Osaka, Japan Introduction: The metabolism of arachidonic acid by either cyclooxygenase or lipoxygenase is believed to play an important role in carcinogenesis. Leukotriene
(LT) D4 is a proinflammmatory mediator derived from arachidonic acid through various enzymatic steps, and 5-lipoxygenase is an important factor in generating LTD4. We investigated LTD4 receptor (cysteinylLT1 receptor: CysLT1R) expression in renal cell carcinoma (RCC), as well as the effects of CysLT1R antagonist on cell proliferation in RCC cell line. Materials and Methods: CysLT1R expression in RCC patients and normal kidney (NK) tissues were examined. CysLT1R expression was detected by immunohistochemistry. Effects of CysLT1R antagonist on RCC cell growth were examined by MTT assay. Flow cytometry was used to determine whether or not the CysLT1R antagonist induces apoptosis. Results: Initially, only slight CysLT1R expression was detected in NK tissues and marked CysLT1R expression was detected in RCC tissues. CysLT1R expression was higher in high-grade cancer than in lowgrade cancer. Furthermore, CysLT1R antagonist caused marked inhibition of RCC cells in a concentration-dependent and time-dependent manner through early apoptosis. Conclusion: CysLT1R is induced in RCC, and the results suggest that CysLT1R antagonist may mediate potent anti-proliferative effects of RCC cells. Thus, the target of CysLT1R may become a new therapy in the treatment of RCC. UP-2.068 Gamma-Aminobutyric Acid Modulates Invasive Property in Renal Cancer Inamoto T1, Azuma H1, Kotake Y1, Ubai T1, Watanabe M2, Katsuoka Y1 1 Dept. of Urology, 2Dept. of Anatomy, Osaka Medical College, Osaka, Japan Introduction and Objective: Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system. In addition, GABA and GABA receptors exist in many non-neuronal peripheral tissues, and serve wide variety of roles including cell division, differentiation, and maturation of cells. We found that GABA facilitated renal cancer invasion. Herein, we show the findings that modulation of GABA receptor alters invasion dynamics in renal cancer. Materials and Methods: To determine GABA receptor expression, Caki-2, VMRCRCW, and ACHN cells were analyzed by FACS. To modulate GABA receptor pathway related with cancer invasion, muscimol (an agonist for GABA-A), baclofen (an agonist for GABA-B), bicuculline methiodide (an antagonist for GABA-A), and CGP 35348 (an antagonist for GABA-B) were utilized. To silence 44/42 MAPK responses, siRNA to
UROLOGY 74 (Supplment 4A), October 2009
ejaculatory duct and prostate. Among all the cases followed up more than 3 to 51 months after operation, 36 patients (81.8%) had improved semen parameters and 11 patients’ wives (25%) had pregnancies in 44 cases of TURED, 6 patients (60%) had improved semen parameters and 3 patients’ wives (30%) had pregnancies in 10 cases of vasoepididymostomy. Conclusion: Semen analyses of fructose and alpha-glucosidase measurement in seminal plasma, TRUS and vasography are major diagnostic methods for obstructive azoospermia. TURED and vasoepididymostomy may separately be effective methods for the treatment of azoospermia with EDO and obstructive azoospermia with epididymal level.
UP-2.065 Expression of GAT1 in Male Reproductive System and Its Effects on Reproduction in Mice Zhang J, Gui Y, Yuan T, Bian C Department of Urology, Tongji Hospital of Tongji University, Shanghai, China Objective: The present study was carried out to identify GABA transport protein I (GAT1) in male reproductive organs and to study the effect of GAT1 overexpression in the male reproductive system of GAT1 transgenic mice (TG). Materials and Methods: Expression and location of GAT1 in testes, epididymis and sperm of wild-type (WT) mice were identified by immunohistochemistry and westernblot. Histological changes of testis, epididymis and sperm of transgenic mice overexpressing GAT1 were detected by immunofluorescent staining and HE staining. Results: GAT1 expression was detected in the testis, epididymis and sperm of non-transgenic mice. Vacuolization and deformity of spermatogenic cells were observed in the transgenic mice, but the epididymis was unremarkable. Immunofluorescent staining showed that the number of diastrophic and decapitated sperm increased significantly in transgenic mice to 46.9% from 7.3% in nontransgenic mice. Conclusion: These results suggest that abnormal expression of GAT1 could result in spermogenesis function injury, sperm paramorphia and dysgenesis.
UP-2.066 Increased Serum Hepcidin-25 Level and Increased Tumor Expression of Hepcidin MRNA is Associated with Metastasis of Renal Cell Carcinoma
S252
Abe H1, Tomosugi N2, Kamai T1, Arai K1, Yoshida K1 1 Department of Urology, Dokkyo Medical University, Tochigi, Japan; 2Proteomics Research Unit, Division of Advanced Medicine, Medical Research Institute, Kanazawa Medical University, Ishikawa, Japan Introduction and Objective: Hepcidin is a molecule that is important for iron metabolism. We investigated the role of hepcidin in renal cell carcinoma (RCC). Materials and Methods: We measured serum hepcidin-25 levels in 32 patients by liquid chromatograpy (LC)-mass spectrometry (MS)/MS, and assessed hepcidin mRNA expression in paired tumor and non-tumor tissue samples from the surgical specimens of 53 consecutive patients with RCC by using a real-time reverse transcription polymerase chain reaction. Results: The serum hepcidin-25 level was higher in metastatic RCC than in nonmetastatic RCC (P⬍0.0001). Expression of hepcidin mRNA was lower in tumor tissue than in non-tumor tissues (P⬍0.0001). The serum hepcidin-25 level was not correlated with the expression of hepcidin mRNA in the corresponding tumor tissue specimens from 32 patients. Hepcidin mRNA expression in tumor tissue was correlated with metastatic potential, but not with tumor differentiation or the local stage. Kaplan-Meier analysis showed that high hepcidin mRNA expression was related to shorter overall survival in all patients. Univariate analysis according to the Cox proportional hazards model showed that hepcidin mRNA level was an independent prognostic factor in overall survival. Conclusions: Our findings suggest that a high serum hepcidin-25 level may be an indicator of the progression of RCC, and that upregulation of hepcidin mRNA in tumor tissue may be related to increased metastatic potential. UP-2.067 The Target of Cysteinyl-Leukotriene1 Receptor (CysLT1R) Is New AntiCancer Strategy for Human Renal Cell Carcinoma Funao K, Matsuyama M, Kuratsukuri K, Tanaka T, Takemoto Y, Nakatani T, Yoshimura R Dept. of Urology, Osaka City University, Graduate School of Medicine, Osaka, Japan Introduction: The metabolism of arachidonic acid by either cyclooxygenase or lipoxygenase is believed to play an important role in carcinogenesis. Leukotriene
(LT) D4 is a proinflammmatory mediator derived from arachidonic acid through various enzymatic steps, and 5-lipoxygenase is an important factor in generating LTD4. We investigated LTD4 receptor (cysteinylLT1 receptor: CysLT1R) expression in renal cell carcinoma (RCC), as well as the effects of CysLT1R antagonist on cell proliferation in RCC cell line. Materials and Methods: CysLT1R expression in RCC patients and normal kidney (NK) tissues were examined. CysLT1R expression was detected by immunohistochemistry. Effects of CysLT1R antagonist on RCC cell growth were examined by MTT assay. Flow cytometry was used to determine whether or not the CysLT1R antagonist induces apoptosis. Results: Initially, only slight CysLT1R expression was detected in NK tissues and marked CysLT1R expression was detected in RCC tissues. CysLT1R expression was higher in high-grade cancer than in lowgrade cancer. Furthermore, CysLT1R antagonist caused marked inhibition of RCC cells in a concentration-dependent and time-dependent manner through early apoptosis. Conclusion: CysLT1R is induced in RCC, and the results suggest that CysLT1R antagonist may mediate potent anti-proliferative effects of RCC cells. Thus, the target of CysLT1R may become a new therapy in the treatment of RCC. UP-2.068 Gamma-Aminobutyric Acid Modulates Invasive Property in Renal Cancer Inamoto T1, Azuma H1, Kotake Y1, Ubai T1, Watanabe M2, Katsuoka Y1 1 Dept. of Urology, 2Dept. of Anatomy, Osaka Medical College, Osaka, Japan Introduction and Objective: Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system. In addition, GABA and GABA receptors exist in many non-neuronal peripheral tissues, and serve wide variety of roles including cell division, differentiation, and maturation of cells. We found that GABA facilitated renal cancer invasion. Herein, we show the findings that modulation of GABA receptor alters invasion dynamics in renal cancer. Materials and Methods: To determine GABA receptor expression, Caki-2, VMRCRCW, and ACHN cells were analyzed by FACS. To modulate GABA receptor pathway related with cancer invasion, muscimol (an agonist for GABA-A), baclofen (an agonist for GABA-B), bicuculline methiodide (an antagonist for GABA-A), and CGP 35348 (an antagonist for GABA-B) were utilized. To silence 44/42 MAPK responses, siRNA to
UROLOGY 74 (Supplment 4A), October 2009