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• W2038305087 abstract "You have accessJournal of UrologyProstate Cancer: Basic Research (IV)1 Apr 2013802 LOSS OF DICKKOPF-3 EXPRESSION IMPAIRS PROSTATE ACINAR MORPHOGENESIS VIA ABERRANT TGF-β/SMAD SIGNALLING ACTIVATION Yoshiaki Kawano, Diana Romero, Nora Bengora, Marjorie Walker, Masatoshi Eto, Jonathan Waxman, Christof Niehrs, and Robert Kypta Yoshiaki KawanoYoshiaki Kawano Kumamoto, Japan More articles by this author , Diana RomeroDiana Romero London, United Kingdom More articles by this author , Nora BengoraNora Bengora Derio, Spain More articles by this author , Marjorie WalkerMarjorie Walker London, United Kingdom More articles by this author , Masatoshi EtoMasatoshi Eto Kumamoto, Japan More articles by this author , Jonathan WaxmanJonathan Waxman London, United Kingdom More articles by this author , Christof NiehrsChristof Niehrs Mainz, Germany More articles by this author , and Robert KyptaRobert Kypta London, United Kingdom More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.366AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To investigate the role of the tumour suppressor Dickkopf-3 in prostate acinar formation in vitro and in vivo. METHODS To establish Dkk-3 depleted RWPE-1 cell lines (RWPE-1/D3-sh cells), RWPE-1 cells were stably transfected with Dkk-3-targeting shRNA. The cells transfected with non-targeting vector (RWPE-1/NS-sh) were used as a control. For 3D acinar morphogenesis assays, either RWPE-1/NS-sh or RWPE-1/D3-sh cells were plated on a thin-layered bed of Matrigel (BD Biosciences) on 8-chamber glass slides and cultured with Keratinocyte SFM (Invitrogen) containing 2% Matrigel. For gene reporter assays, cells were transfected with luciferase and bet-galactosidase-based reporters. For analysis of Dkk-3 knockout mice and their wild-type littermates, prostates were dissected from 6-8 week-old mice, fixed with 4% paraformaldehyde and embedded in paraffin. Sections were analysed by H&E and immunofluorescent staining. RESULTS Analysis of prostates from mice showed an increased Ki-67 index in all lobes of Dkk-3 knockout mice, compared to wild type. H&E and immunofluorescent staining for ZO-1 and E-Cadherin revealed subtle changes in cellular structure during prostate development. In 2D culture, there were no apparent morphological differences between RWPE-1/NS-sh cells and RWPE-1/D3-sh cells. However, in 3D assays, while RWPE-1/NS-sh cells underwent normal acinar morphogenesis with spherically arranged polarisation, RWPE-1/D3-sh cells formed disorganised cell aggregates. Immunofluorescent staining for phosphotylated histone H3 revealed increased cell division in Dkk-3 depleted acini. Gene reporter assays and biochemical analysis indicated aberrant activation of TGFβ/smad signalling in RWPE-1/D3-sh cells. Furthermore, SB431542, a specific TGFβ type I receptor inhibitor, rescued the phenotype manifested by RWPE-1/D3-sh cells. These results indicate that Dkk-3 knockdown results in change in TGFβ/smad signalling pathway, abnormal cell division and disorder of cellular polarisation, leading to the disruption of acini. CONCLUSIONS These results suggest that loss of Dkk-3 expression contributes to prostate cancer development due to abnormal cell division and impaired cellular structure by aberrant TGFβ/Smad signalling pathway activation. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e330 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yoshiaki Kawano Kumamoto, Japan More articles by this author Diana Romero London, United Kingdom More articles by this author Nora Bengora Derio, Spain More articles by this author Marjorie Walker London, United Kingdom More articles by this author Masatoshi Eto Kumamoto, Japan More articles by this author Jonathan Waxman London, United Kingdom More articles by this author Christof Niehrs Mainz, Germany More articles by this author Robert Kypta London, United Kingdom More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ..." @default.
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• W2038305087 title "802 LOSS OF DICKKOPF-3 EXPRESSION IMPAIRS PROSTATE ACINAR MORPHOGENESIS VIA ABERRANT TGF-β/SMAD SIGNALLING ACTIVATION" @default.
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