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INTRODUCTION AND OBJECTIVES: The Wnt signaling pathway (canonical and non-canonical) regulates crucial aspects of embryonic development. Previously, we have reported that non-canonical Wnt members promote proliferation of cancer cells by increasing secretion of growth factors and expression of cancer related genes. We have also demonstrated that Wnt5a is predominantly expressed in prostatic stromal cells and plays a key role in stromalepithelial communication and in the development of prostate cancer. The purpose of this study is to reveal the relation between the noncanonical Wnt signaling and AR signaling on prostate. METHODS: Prostate tissue samples were obtained from patients undergoing radical prostatectomy or cystectomy for prostate and bladder cancer respectively. Cells, representing the cancer, stromal from around the cancer, as well as epithelium and stromal from pathologically normal tissues were isolated by laser capture microdissection Stromal cells (WPMY1 and PrSC) were cultured with DHT ligand and protein. The mRNA levels of Wnt5a, AR, and PSA were determined in both sample types. Wnt5a-knockdown in WPMY1 cells (shWnt5a) along with a vector control (shCTL) was performed and effects on growth evaluated by MTT assay. The cells were co-cultured with PC3 cells, and the mRNA expression of AR was evaluated. A DNA fragment upstream of the Wnt5a gene was inserted into pGL4.17 a vector and the luciferase activity representing Wnt5a inductionby DHT was measured. RESULTS: Wnt5a mRNA expression was prediminanly in the stromal cells of the human prostate samples and the expression levels observed were in proportion to PSA mRNA expression levels. DHT increased production of AR as well as PSA and Wnt5a mRNA expression in both WPMY1 and PrSC cells. Wnt5 knockdown decreased the proliferation of WPMY1cells by 50% compared to shCTR. Co-culture of PC3 cells and WPMY1shWnt5a formed fewer and smaller sized colonies than co-culture of PC3 cells and WPMY1shCTR. AR and PSA mRNA expression levels were remarkably lower in WPMY1shWnt5a than in the control. DHT ligand dramatically increased the luciferase activity of Wnt5a promoter in WPMY1 cells. CONCLUSIONS: The AR is expressed not only in prostatic epithelium but also prostatic stromal as measured by their response to DHT. In stromal cells, The studies presented here demonstrated an interplay between AR signaling and Wnt5a that appears to modulate the proliferation of prostate cancer cells providing further support for the importance of Wnt5a in the regulation of prostate cancer.

Introduction. Erectile dysfunction (ED) is a major health problem. It is known that diabetic patients are more refractory to common treatments for ED. Aim. To explore the better treatment for ED, we examined the effects of adipose-derived stem cells (ASC) on ED using a diabetic rat model. We also analyzed the cytokines produced by ASC and implicated in ASC-induced restoration of erectile function. Methods. Male Wistar rats were injected with streptozotocin (STZ) to induce diabetes. ASC or adenoviruses were injected into the penis 6 weeks after STZ administration. Erectile function, penile histology and protein expression were analyzed 4 weeks after the injection of ASC or adenoviruses. Main Outcome Measures. Intracavernous pressure and mean arterial pressure were measured to evaluate erectile function. The morphology of the penis was analyzed by Elastica van Gieson stain and immunohistochemistry. The expression of proteins specific for vascular endothelial cells (VEC) was assessed by Western blot analysis. Results. ASC restored erectile function especially when they were cultured in medium containing growth factors for VEC. This restoration was associated with improvement in the histology of the cavernous body, and increased expression of VEC markers such as VE-cadherin and endothelial nitric oxide synthase (eNOS). When the expression of adrenomedullin (AM), a vasoactive peptide originally isolated from human pheochromocytoma tissue, was knocked down, the effect of ASC on ED was significantly diminished. Knockdown of AM was associated with decreased expressions of VE-cadherin and eNOS. Furthermore, overexpression of AM induced by adenovirus infection significantly improved erectile function in these diabetic rats. Overexpression of AM was associated with increased expressions of VE-cadherin and eNOS. Conclusions. These results suggested that ASC have the potentials to restore erectile function and that AM produced by ASC plays a major role in the restoration of erectile function. Nishimatsu H, Suzuki E, Kumano S, Nomiya A, Liu M, Kume H, and Homma Y. Adrenomedullin mediates adipose tissue-derived stem cell-induced restoration of erectile function in diabetic rats. J Sex Med **;**:**-**.

INTRODUCTION AND OBJECTIVES: Tumor suppressor CADM1 (cell adhesion molecule 1) belongs to the immunoglobulin-superfamily cell adhesion molecules (IgCAMs) and participates in cell-cell adhesion activity in various epithelia, including those of urinary bladder. On the other hand, CADM1 is frequently inactivated in a variety of human cancers chiefly by gene promoter methylation, especially in their advanced stages. In this study, we investigated possible involvement of CADM1 in bladder cancer. METHODS: CADM1 expression in 11 human bladder cancer cell lines was examined by RT-PCR analysis and western blotting analysis. The methylation status of the CADM1 gene promoter in bladder cancer cell lines was examined by pyrosequencing analysis. CADM1 expression in 147 human primary bladder cancers was examined by immunohistochemical analysis. Cadm1-deficient mice as well as the wild-type C57BL/6 mice were administrated 0.05% N-butyl-N(4-hydroxybutyl)nitrosamine (BBN), an established bladder carcinogen for mice, in the drinking water for 20 weeks. Any human examination was conducted according to the protocol approved by the research ethics committee of Graduated School of Medicine, The University of Tokyo. RESULTS: CADM1 expression was absent or greatly reduced in 6 of 11 human bladder cancer cell lines. The promoter methylation of the CADM1 gene was observed in 4 of those 6 cell lines. CADM1 was expressed along the cell membrane in normal urothelium of human bladder. On the other hand, membranous expression was lost or altered in 92 (63%) of 147 primary bladder cancers. Loss or aberrant expression of CADM1 was significantly correlated with higher pT stages (pTa pT1 pT2-4), higher nuclear grade (G1-2 G3). Moreover, loss or aberrant expression was closely correlated with poor prognosis (P 0.001) and was an independent prognostic factor in multivariate analysis (P 0.021) in 112 patients treated with radical cystectomy. Administration of BBN developed invasive bladder cancers (pT2-4) at significantly higher incidence in Cadm1-deficient mice than in the wild-type mice (P 0.025). CONCLUSIONS: The disruption of CADM1 would be involved in the progression of bladder cancers, especially in the invasion and metastasis, and could provide a prognostic marker in invasive bladder cancer. Source of Funding: None