Yasuyo Yamamoto

Distinct roles of Rab3B and Rab13 in the polarized transport of apical, basolateral, and tight junctional membrane proteins to the plasma membrane.

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.

Expression of angiopoietin‐1 and ‐2, and its clinical significance in human bladder cancer

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.

Laparoscopic nephroureterectomy for dysplastic kidney in children: an initial experience

Background: Laparoscopic nephroureterectomy for dysplastic kidney is now becoming a widely accepted procedure. We report here our initial experience with laparoscopic nephroureterectomy in four girls.

Functional evaluation of one‐stage urethroplasty with parameatal foreskin flaps repair of hypospadias using uroflowmetry

Background: 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.

Complications of Flexible Ureteroscopic Treatment for Renal and Ureteral Calculi during the Learning Curve

Background: The flexible ureterorenoscope (URS) and associated devices have developed rapidly. However, despite its therapeutic benefits, URS may be associated with some complications. To the best of our knowledge, there are no studies discussing the complications of flexURS during the learning curve. Methods: A retrospective review of the records of patients who underwent flexURS from January 2005 to June 2013 was performed. To compare the complications after the introduction of flexURS, patients were divided into four groups based on the surgeons training experience, that is, based on the number of cases performed by the surgeon. A total of 219 cases underwent flexURS. Groups 1, 2, 3, and 4 included 35, 50, 50, and 84 cases, respectively. The complications were classified using the Clavien system (I-IV). Results: The mean operation time and stone-free rate were significantly different (p < 0.001, p = 0.013, respectively). The total complication rates were 13.6, 10, 8.3, and 3.2%, respectively (p = 0.068). The more the surgeons experience, the less was the complication rate. Despite our best efforts, the incidence of urosepsis was not reduced (p = 0.902). Conclusions: To reduce severe complications, it is necessary to have performed about 100 cases. Increased surgeon experience tended to decrease the risk of severe complications, but the incidence of urosepsis was not reduced.

Clinical Significance of Neoadjuvant Combined Androgen Blockade for More Than Six Months in Patients with Localized Prostate Cancer Treated with Prostate Brachytherapy.

Introduction: The aim of this study is to clarify the clinical significance of neoadjuvant combined androgen blockade (CAB) for ≥6 months in patients with localized prostate cancer. Patients and Methods: A total of 431 patients with localized prostate cancer who underwent prostate brachytherapy (BT) with or without neoadjuvant CAB for ≥6 months with mean follow-up time of 64.6 months (range 24-108 months) were evaluated retrospectively. Of those 431, 232 patients received BT in combination with neoadjuvant CAB for ≥6 months. Biochemical recurrence-free rates (BRFRs) in 364 patients with at least 3 years of follow-up were evaluated by log-rank test. Results: BRFR in patients with low-, intermediate- and high-risk prostate cancer were 98.1, 94.2 and 89.1%, respectively. In patients with intermediate-risk prostate cancer only, neoadjuvant CAB was significantly associated with BRFR (p = 0.0468). Especially in patients with intermediate-risk prostate cancer with radiation dose received by 90% of the prostate (D90) <180 Gy, neoadjuvant CAB exerted a favorable impact on BRFR (p = 0.0429). On multivariate analyses, neoadjuvant CAB and D90 were independent predictors of BRFR (p = 0.0061 and p < 0.0001, respectively). Conclusions: Neoadjuvant CAB for ≥6 months has a favorable impact on BRFR in patients with intermediate-risk prostate cancer, particularly in patients with relatively low radiation doses of D90.

MP72-06 CORRELATION OF THE INSULIN RECEPTOR EXPRESSION AND THE RESISTANCE TO VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR TYROSINE KINASE INHIBITORS IN ADVANCED CLEAR CELL RENAL CELL CARCINOMA

INTRODUCTION AND OBJECTIVES: Vascular endothelial growth factor receptor tyrosine kinase inhibitors (VEGFR-TKIs) demonstrate the significant efficacy for advanced clear cell renal cell carcinoma (ccRCC), however, it eventually becomes resistant to VEGFR-TKIs during the treatment. So far, the mechanisms for the resistance to VEGFR-TKIs remain to be fully elucidated. Previously we have identified the gene set which could predict poor prognosis of ccRCC patients (Takahashi M et.al., Proc Natl Acad Sci U S A., 98: 9754, 2001). We examined whether the insulin receptor (INSR) expression in the gene set may correlate with the resistance to VEGFR-TKIs. METHODS: The INSR expression was examined immunohistochemically in the nephrectomy specimens of the RCC patients (n1⁄433) who then received axitinib as the VEGFR-TKI and correlated with their survival outcome. We compared the INSR expression of the nephrectomy or metastasectomy specimens before and after the administration of VEGFR-TKIs in 5 cases. The INSR expression of the human renal glomerular endothelial cells (HGEC) was compared before and after the administration of axitinib by Western blotting. In addition, we established patient derived Xenograft model (PDX) of ccRCC. Tumors of PDX were resected when it regrew and showed the resistance for axitinib and the INSR expression was compared before and after the treatment by Western blotting. RESULTS: The INSR was expressed at the vessels surrounding tumor cells. Progression-free survival (PFS) was significantly shorter in the INSR-negative group than in the INSR-positive group. Multivariate analysis revealed that the INSR expression was the significantly independent predictor of PFS. In the specimens resected after VEGFR-TKI, the INSR expression was more frequently decreased. As the concentration of axitinib increased, the INSR expression in the HGEC was decreased. The tumors of PDX that were resected after demonstrating the resistance to axitinib had the decreased INSR expression. CONCLUSIONS: The decreased INSR expression could be correlated with the resistance to VEGFR-TKI and its expression may be useful in selecting appropriate drugs for advanced ccRCC patients. Source of Funding: None

MP45-17 GALECTIN-3 PLAYS CRITICAL ROLES FOR THE GROWTH OF BENIGN PROSTATIC HYPERPLASIA

INTRODUCTION AND OBJECTIVES: Galectin-3, a multifunctional oncogenic protein, has been reported to play important roles of progression in a variety of cancer including prostate cancer through the regulation of cancer cell proliferation, apoptosis, invasion and metastasis. However, we also identified the frequent up-regulation of Galectin-3 in benign prostatic hyperplasia (BPH), yet its pathophysiological roles in BPH. Here, we report the involvement of Galectin-3 in the growth of BPH. METHODS: To investigate the association of Galectin-3 expression and prostate volume, we examined serum Galectin-3 (pg/ ml) with ELISA method in non-cancer cohort and analyzed the correlation between Galectin-3 expression and prostate volume with Speamans correlation coefficient. Next, to analyzed proliferation abilities and the effect of Galectin-3 on smooth muscle, we examined knockdown of Galectin-3 expression by siRNA in BPH1 cells (benign prostatic hyperplasia cell line) and a co-culture experiment of BPH1 and PrSMC (Normal Human Prostate Smooth Muscle. Cells). Moreover, to investigate biological function of Galectin-3, we examined the gene expression profiles in Galectin-3-depleted BPH1 cells with microarray and bioinformatics analyses. RESULTS: Correlation analysis revealed that serum Galectin-3 (ng/ml) were correlated with prostate volume (R1⁄40.643 p1⁄40.023) (Figure 1). Next, depletion of Galectin-3 suppressed cell proliferation in BPH1 cells. Moreover, depletion of Galectin-3 in BPH1 cells suppressed cell proliferation of PrSMC cells in a co-culture method, suggesting Galectin-3 enhanced proliferation of PrSMC cells. Bioinformatics analysis with GSEA revealed that depletion of Galectin-3 was involved in interferon a response and interferon a response and interferon ? response (FDR q value < 0.001) (Figure 2), suggesting Galectin-3 regulates the proliferation of PrSMC through interferon response. CONCLUSIONS: Our findings suggest that Galectin-3 is significantly involved in the growth of BPH and a promising therapeutic target for patients with BPH. Source of Funding: GSK Japan Research Grant 2016

Substitution of anti-androgens and tegafur-uracil combination therapy for castration-resistant prostate cancer: Results of a multi-center randomized phase II study

We conducted this study to determine whether substitution with anti-androgen (SOA) and tegafur-uracil (a pro‑drug of 5-FU) combination therapy is more effective than SOA alone after relapse from initial hormonal therapy. Patients who were histologically confirmed and relapsed after initial hormonal therapy were included. All patients were randomly allocated into two groups: SOA alone (group A) or SOA combined with tegafur-uracil (group B). The mRNA expression of four enzymes, including thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), orotate phospho-ribosyltransferase (OPRT) and thymidine phosphorylase (TP), in prostate cancer cells was analyzed by quantitative reverse-transcription polymerase chain reaction. Fifty-two patients were enrolled in this study. The median age was 77 (range: 47-92) years. The PSA response rate in group B (61.5%) tended to be higher compared to that in group A (34.6%) (p=0.095). Group B (median: 15.9 months) had a significantly longer time to PSA progression (TTP) compared to group A (6.4 months) (p=0.014). In patients with a lower TS expression or a higher OPRT expression, group B demonstrated a higher PSA response rate compared to group A (p=0.019 and p=0.041, respectively). In addition, in the patients with a lower TS expression, group B demonstrated a significantly longer TTP compared to group A (p=0.018). There were no severe adverse events in either treatment group. After relapse from initial hormonal therapy, SOA combined with tegafur-uracil is effective and well tolerated. The TS mRNA expression level may be a predictive factor for this combination therapy.

627 ENHANCEMENT OF OSTEOCLASTOGENIC ACTIVITY IN OSTEOLYTIC PROSTATE CANCER CELLS BY PHYSICAL CONTACT WITH OSTEOBLASTS

INTRODUCTION AND OBJECTIVES: Low-temperature sensitive liposomes (LTSLs) release their encapsulated drug into targeted tissue when activated by a source of hyperthermia. The efficacy of an LTSL formulation of docetaxel (DOC) or doxorubicin (DOX) was compared against prostate cancer in a xenograft mouse model. METHODS: Under an approved IACUC protocol, Luciferase transfected human prostate PC-3M-luciferase cells were inoculated (3 10 6 cells) subcutaneously in the right hind leg of 8 wk old female athymic nude mice. When tumors reached a volume of 200–300 mm 3, mice were randomized to receive one intravenous injection of saline, Stealth liposomal DOX (5 mg/kg), LTSL DOX (5 mg/kg), or LTSL DOC (15 mg/kg), with or without hyperthermia treatment (LTSL DOX and LTSL DOC were supplied by Celsion Corp., Columbia, MD). Mice undergoing hyperthermia treatment were anesthetized and stabilized in a holder that allowed for only the leg with tumor to be heated to 41–42C that triggered LTSL drug release. Mice were monitored daily for tumor volume and body weight. Study end-points included growth of tumor to 5x the initial treatment volume or monitoring of survival for 60 days. RESULTS: The LTSL DOC delayed tumor growth longer (20 days) than DOX (7 days) or LTSL DOX (9 days) with hyperthermia (P 0.05). Mice treated with LTSL DOC and hyperthermia survived longest (60 days) compared to all other mice (range 6–8 days, P 0.05). LTSL in the setting of hyperthermia demonstrated complete regression of tumor in 57% of mice. CONCLUSIONS: LTSL DOC with hyperthermia delayed tumor growth more than all other treatments. Survival studies suggest LTSL DOC is a more effective temperature sensitive delivery system against PC-3M prostate tumors.