MP54-06 THREE-DIMENSIONAL ORGANOID CULTURE REVEALS INVOLVEMENT OF WNT/β-CATENIN PATHWAY IN PROLIFERATION OF BLADDER CANCER CELLS

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I1 Apr 2018MP54-06 THREE-DIMENSIONAL ORGANOID CULTURE REVEALS INVOLVEMENT OF WNT/β-CATENIN PATHWAY IN PROLIFERATION OF BLADDER CANCER CELLS Takahiro Yoshida, Nikolai Sopko, Max Kates, Xiaopu Liu, Gregory Joice, David McConkey, and Trinity Bivalacqua Takahiro YoshidaTakahiro Yoshida More articles by this author , Nikolai SopkoNikolai Sopko More articles by this author , Max KatesMax Kates More articles by this author , Xiaopu LiuXiaopu Liu More articles by this author , Gregory JoiceGregory Joice More articles by this author , David McConkeyDavid McConkey More articles by this author , and Trinity BivalacquaTrinity Bivalacqua More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1697AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES There has been increasing awareness of the importance of three-dimensional culture of cancer cells. Tumor cells growing as multicellular spheroids (organoids) are believed to more closely mimic solid tumors in situ. Meanwhile, Wnt/β-catenin pathway was reported to be upregulated in human bladder cancer specimens. However, no clear evidence has been reported that the pathway is directly involved in proliferation of bladder cancer cells. In this study, we assessed the involvement of Wnt/β-catenin pathway in proliferation of bladder cancer cells using organoid culture. METHODS Organoids from bladder cancer cell lines RT4 (luminal phenotype) and 5637 (basal phenotype) were generated by an aggregation method. A partial-digestion method was applied to prepare organoids directly from patient bladder cancer specimens. Wnt/β-catenin pathway was activated by using a small molecule CHIR99021 (GSK3 inhibitor, Wnt activator) and inhibited by siRNA against β-catenin. Activation of Wnt/β-catenin pathway was confirmed by upregulated AXIN2 mRNA expression and translocation of β-catenin into nucleus. Proliferation of cancer cells were evaluated by growth assay under microscope and ATP viability assay. Differentiation status of organoids over growth was characterized by qRT-PCR and western blot. RESULTS CHIR99021 inhibited proliferation of cell lines in conventional monolayer adherent culture, but promoted proliferation in three-dimensional organoid culture with activation of Wnt/β-catenin pathway. Enhanced proliferation of cancer cells with activation of Wnt/β-catenin pathway by CHIR99021 was also observed in patient-derived organoids. When β-catenin was knockdowned in cell lines, the growth of organoids was significantly suppressed. Cytokeratin 20, a terminal differentiation marker, was less expressed over CHIR99021-enhanced cell proliferation. CONCLUSIONS We showed for the first time that Wnt/β-catenin pathway was directly involved in proliferation of bladder cancer cells, suggesting that Wnt/β-catenin pathway may be a potential target for the treatment of a subset of bladder cancer. The above finding could not be observed when the same cells were grown in conventional two-dimensional adherent cultures, providing a concrete example of why organoid culture is important for cancer research. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e714 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Takahiro Yoshida More articles by this author Nikolai Sopko More articles by this author Max Kates More articles by this author Xiaopu Liu More articles by this author Gregory Joice More articles by this author David McConkey More articles by this author Trinity Bivalacqua More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...