Title
278
Androgen receptor positive stromal cells regulate prostate cancer proliferation through noncanonical Wnt signaling Eur Urol Suppl 2015;14/2;e278
Print! Print!
Takahashi S. 1 , Takada I.2 , Terada N.3 , Getzenberg R.H.4 , Homma Y. 1 1 Graduate
School of Medicine, The University of Tokyo, Dept. of Urology, Tokyo, Japan, 2 Nihon University School of Medicine, Dept. of
Biomedical Sciences, Tokyo, Japan, 3 The University of Kyoto, Dept. of Urology, Tokyo, Japan, 4 GTx Inc., Memphis, United States of America INTRODUCTION & OBJECTIVES: The Wnt signaling pathway (canonical and non-canonical) regulates crucial aspects of embryonic development. Previously, we have reported that non-canonical Wnts promotes proliferation of cancer cells by increasing secretion of growth factors and expression of cancer related genes. We have also proved that Wnt5a expresses mainly in stromal cells of prostate and plays a key role in stromal-epithelial communication and in the development of prostate cancer. The role of androgen receptor (AR) in prostatic stromal is not known, yet. The purpose of this study is to reveal the relation between the non-canonical Wnt signaling and AR signaling on prostate. MATERIAL & METHODS: Prostate samples were obtained from patients with prostate and bladder cancer and 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 and the mRNA levels of Wnt5a, AR, and PSA. Stromal cells (WPMY1 and PrSC) were cultured with DHT ligand and protein or mRNA expression of AR, PSA and Wnt5a were measured. Then,Wnt5a-knockdown in WPMY1 cells (shWnt5a) along with a vector control (shCTL) were established, MTT assay and colony assay of co-culture with PC3 cells were performed, and the mRNA expression of AR was evaluated. Finally, reporter assay of Wnt5a was performed. Human DNA fragment of upstream of Wnt5a gene was inserted into pGL4.17 vector and the luciferase activity in WPMY1 cells induced by DHT was measured. RESULTS: Wnt5a mRNA was mainly expressed in stromal cells on human prostate samples and the expression levels were in proportion with PSA mRNA expression levels which is target gene of AR. DHT ligands increased production of AR protein and also PSA and Wnt5a mRNA expression in both WPMY1 and PrSC cells. The proliferation of WPMY1shWnt5a was suppressed by 50% than shCTR by MTT assay. Co-culture of PC3 cells and WPMY1shWnt5a formed less numbers and smaller size of colonies than co-culture of PC3 cells and WPMY1shCTR. AR and PSA mRNA expression levels were remarkable lower in WPMY1shWnt5a than in control. DHT ligand dramatically increased the luciferase activity of Wnt5a promoter in WPMY1 cells. CONCLUSIONS: ARs express not only in prostatic epithelium but also prostatic stromal by low concentration and response to DHT. In stromal cells, AR-DHT signal modulates Wnt5a expression and the Wnt5a promotes excretion of growth factors from stromal cells which stimulate prostate cancer cells proliferation.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/278.html[19/02/2015 08:10:10]
278
Androgen receptor positive stromal cells regulate prostate cancer proliferation through noncanonical Wnt signaling Eur Urol Suppl 2015;14/2;e278
Print! Print!
Takahashi S. 1 , Takada I.2 , Terada N.3 , Getzenberg R.H.4 , Homma Y. 1 1 Graduate
School of Medicine, The University of Tokyo, Dept. of Urology, Tokyo, Japan, 2 Nihon University School of Medicine, Dept. of
Biomedical Sciences, Tokyo, Japan, 3 The University of Kyoto, Dept. of Urology, Tokyo, Japan, 4 GTx Inc., Memphis, United States of America INTRODUCTION & OBJECTIVES: The Wnt signaling pathway (canonical and non-canonical) regulates crucial aspects of embryonic development. Previously, we have reported that non-canonical Wnts promotes proliferation of cancer cells by increasing secretion of growth factors and expression of cancer related genes. We have also proved that Wnt5a expresses mainly in stromal cells of prostate and plays a key role in stromal-epithelial communication and in the development of prostate cancer. The role of androgen receptor (AR) in prostatic stromal is not known, yet. The purpose of this study is to reveal the relation between the non-canonical Wnt signaling and AR signaling on prostate. MATERIAL & METHODS: Prostate samples were obtained from patients with prostate and bladder cancer and 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 and the mRNA levels of Wnt5a, AR, and PSA. Stromal cells (WPMY1 and PrSC) were cultured with DHT ligand and protein or mRNA expression of AR, PSA and Wnt5a were measured. Then,Wnt5a-knockdown in WPMY1 cells (shWnt5a) along with a vector control (shCTL) were established, MTT assay and colony assay of co-culture with PC3 cells were performed, and the mRNA expression of AR was evaluated. Finally, reporter assay of Wnt5a was performed. Human DNA fragment of upstream of Wnt5a gene was inserted into pGL4.17 vector and the luciferase activity in WPMY1 cells induced by DHT was measured. RESULTS: Wnt5a mRNA was mainly expressed in stromal cells on human prostate samples and the expression levels were in proportion with PSA mRNA expression levels which is target gene of AR. DHT ligands increased production of AR protein and also PSA and Wnt5a mRNA expression in both WPMY1 and PrSC cells. The proliferation of WPMY1shWnt5a was suppressed by 50% than shCTR by MTT assay. Co-culture of PC3 cells and WPMY1shWnt5a formed less numbers and smaller size of colonies than co-culture of PC3 cells and WPMY1shCTR. AR and PSA mRNA expression levels were remarkable lower in WPMY1shWnt5a than in control. DHT ligand dramatically increased the luciferase activity of Wnt5a promoter in WPMY1 cells. CONCLUSIONS: ARs express not only in prostatic epithelium but also prostatic stromal by low concentration and response to DHT. In stromal cells, AR-DHT signal modulates Wnt5a expression and the Wnt5a promotes excretion of growth factors from stromal cells which stimulate prostate cancer cells proliferation.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/278.html[19/02/2015 08:10:10]
