501 SONIC HEDGEHOG SIGNALING IS INVOLVED IN EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) IN HUMAN PROSTATE CANCER ARCAP CELLS

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research (III)1 Apr 2013501 SONIC HEDGEHOG SIGNALING IS INVOLVED IN EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) IN HUMAN PROSTATE CANCER ARCAP CELLS Guodong Zhu, Jiancheng Zhou, Haiyen E. Zhau, Linlin Zhang, Wenbin Song, Lei Li, Dalin He, and Leland W.K. Chung Guodong ZhuGuodong Zhu Xi'an, China, People's Republic of More articles by this author , Jiancheng ZhouJiancheng Zhou Xi'an, China, People's Republic of More articles by this author , Haiyen E. ZhauHaiyen E. Zhau Los Angeles, CA More articles by this author , Linlin ZhangLinlin Zhang Xi'an, China, People's Republic of More articles by this author , Wenbin SongWenbin Song Xi'an, China, People's Republic of More articles by this author , Lei LiLei Li Xi'an, China, People's Republic of More articles by this author , Dalin HeDalin He Xi'an, China, People's Republic of More articles by this author , and Leland W.K. ChungLeland W.K. Chung Los Angeles, CA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1895AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Prostate cancer (PCa) progression from localized to disseminated bone and soft tissue metastases have been well characterized using ARCaPE to ARCaPM transition model. In this study, we further examined the role of sonic hedgehog (Shh) signaling activation upon the progression of ARCaP cells, with particular emphasis on their transition from epithelial to mesenchymal phenotype, as evidenced by cell morphology and gene expression profiles. METHODS The Shh expressed ARCaPE cell (ARCaPE-Shh) was established by lipofectamine transfection system. The expression of EMT markers (E-cadherin, CK18, CK19, N-cadherin and Vimentin) was examined in vector control ARCaPE (ARCaPE-Neo), ARCaPE-Shh, and ARCaPM by western blot. The migration and invasion of ARCaP cells, and the effect of cyclopamine (Cycl) on EMT were evaluated respectively by wound healing and trans-well assays. Zinc finger E-box Binding homeobox-1(ZEB-1), Slug, Snail, Gli-1, and Matrix Metallopeptidase 9 (MMP-9) protein expressions were detected by western blot. The biochemical characterizations of Shh signaling components in ARCaP EMT model were correlated with the growth of tumor xenografts in nude mice. Specifically, the expressions of E-cadherin, Vimentin, ZEB-1 and MMP-9 were analyzed in tumor xenografts by immunohistochemistry (IHC). RESULTS The ARCaPE-Shh, when compared to ARCaPE-Neo, exhibited spindle-like scattering morphology, and expressed higher levels of mesenchymal makers (Vimentin and N-cadherin) but lower levels of epithelial makers (E-cadherin, CK18 and CK19); these mesenchymal transition phenotypes coincided with increased cell migration and invasion capability of ARCaPE-Shh cells. The involvement of Shh signaling in ARCaP EMT was further supported by the observation that Cycl effectively inhibited the migratory and invasive capability of ARCaPE-Neo, ARCaPE-Shh and ARCaPM cells. Constitutive overexpression of GLI-1 increased MMP9 and ZEB-1 expression in ARCaPE-Shh cells and these increases were blocked by the simultaneous treatment with Cycl. Increased Shh signaling was also supported by in vivo studies. The average tumor xenograft size in ARCaPE-Shh group was 18±3 mm3 in comparison to ARCaPE-Neo group, 8±2mm3, and the average tumor formation latency for ARCaPE-Shh group was 38±5 days, shorter than ARCaPE-Neo group, 56±6 days. CONCLUSIONS Shh signaling may play an important role in initiation of EMT in PCa cells. Shh signaling inhibitor, Cycl, could be further developed as a therapeutic agent against PCa progression by targeting Shh-induced EMT. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e205-e206 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Guodong Zhu Xi'an, China, People's Republic of More articles by this author Jiancheng Zhou Xi'an, China, People's Republic of More articles by this author Haiyen E. Zhau Los Angeles, CA More articles by this author Linlin Zhang Xi'an, China, People's Republic of More articles by this author Wenbin Song Xi'an, China, People's Republic of More articles by this author Lei Li Xi'an, China, People's Republic of More articles by this author Dalin He Xi'an, China, People's Republic of More articles by this author Leland W.K. Chung Los Angeles, CA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...