1408 INDUCTION OF SECRETED WNT FACTOR WISP1/CCN4 BY HUMAN PROSTATE STROMAL CELLS BY MICROENVIORNMENTAL HYPOXIA

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 20101408 INDUCTION OF SECRETED WNT FACTOR WISP1/CCN4 BY HUMAN PROSTATE STROMAL CELLS BY MICROENVIORNMENTAL HYPOXIA Matthew Tanner, Mengqian Chen, Elina Levina, Richard Carkner, and Ralph Buttyan Matthew TannerMatthew Tanner More articles by this author , Mengqian ChenMengqian Chen More articles by this author , Elina LevinaElina Levina More articles by this author , Richard CarknerRichard Carkner More articles by this author , and Ralph ButtyanRalph Buttyan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1100AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Wnt is a ligand-activated cell signaling pathway that plays an important role in normal embryonic development. The effects of Wnt in development are often manifest through a paracrine signaling process wherein Wnt ligands are secreted by one cell type but act on a neighboring cell type. For prostate epithelial cells, Wnt regulates androgen receptor expression and promotes cell growth under low androgen conditions so Wnt activity mediated by age-associated microenvironmental changes has the potential to effect abnormal prostate growth. We sought to determine whether hypoxia, a microenvironmental condition found in aging prostates, affects the expression of Wnt ligands and downstream Wnt targets in human prostate stromal cells as a means to better understand how this condition might contribute to human prostate disease. METHODS Cultured immortalized human prostate stromal cells, WPMY-1 and PS-30, were incubated under normoxic or hypoxic (1% O2) conditions. Hypoxia responsiveness was assessed by the activity of a hypoxia-sensitive luciferase reporter transfected into the cells. The effects of hypoxia on gene expression were determined by comparative expression analysis of RNAs extracted from normoxic or hypoxic cells using the SABiosciences Wnt-targeted gene microarray expression profiling system and the results were confirmed by real-time quantitative reverse-transcriptase PCR. Human WISP1/CCN4 cDNA was cloned into a lentiviral expression vector and used to produce WISP1/CCN4 overexpressing WPMY-1 cells. RESULTS Hypoxia-sensitive luciferase expression peaked at 72 hrs incubation in 1% O2. Several Wnt-related genes were found to be differentially upregulated by 72 hrs of hypoxia, most prominently WISP1/CCN4 (>50 fold), Wnt-1 (>3-fold), Wnt-16 (>2-fold) and WIF1 (>2-fold). Detailed analysis of the entire WISP/CCN gene family showed that expression of all 3 homologues were highly upregulated by hypoxia. Human cDNA encoding WISP1 was cloned and expressed in WPMY-1 cells to provide a cell model to test for the effects of stromal-expressed WISP1 on prostate epithelial cells. CONCLUSIONS Human prostate stromal cells respond to hypoxia by altering the expression of certain genes in the Wnt pathway. This was especially pronounced for WISP1/CCN4, a Wnt-inducible, growth enhancing gene product that is secreted into the extracellular matrix. The availability of a prostate stromal cell model that overexpresses WISP1/CCN4 will enable studies to test how this protein affects the growth of neighboring benign or malignant epithelial cells. Albany, NY© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e543-e544 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Matthew Tanner More articles by this author Mengqian Chen More articles by this author Elina Levina More articles by this author Richard Carkner More articles by this author Ralph Buttyan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...