MP21-01 EFFECTS OF SILIBININ AGAINST BLADDER CANCER METASTASIS: ROLES OF GSK3β/β-CATENIN/ZEB1 SIGNALING

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research I1 Apr 2014MP21-01 EFFECTS OF SILIBININ AGAINST BLADDER CANCER METASTASIS: ROLES OF GSK3β/β-CATENIN/ZEB1 SIGNALING Kai-jie Wu, Jian-cheng Zhou, Zhong-yun Ning, Jin Zeng, Jin-hai Fan, and Da-lin He Kai-jie WuKai-jie Wu More articles by this author , Jian-cheng ZhouJian-cheng Zhou More articles by this author , Zhong-yun NingZhong-yun Ning More articles by this author , Jin ZengJin Zeng More articles by this author , Jin-hai FanJin-hai Fan More articles by this author , and Da-lin HeDa-lin He More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.829AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Metastatic bladder cancers have a poor prognosis even after cisplatin-based chemotherapy; therefore, novel adjuvant agents with complete efficacy and low toxicity are desired. Our and other previous studies have demonstrated that silibinin, a nontoxic natural flavonoid, exhibited pleiotropic anticancer effects in bladder cancer. It induced apoptosis and inhibited proliferation of bladder cancer cells; however, whether it could suppress bladder cancer metastasis has not been elucidated. METHODS A well-developed highly metastatic T24-L cell model was utilized to study the anti-metastatic effects of silibinin on bladder cancer. Transwell migration and Matrigel invasion assays were employed to investigate the effects of silibinin on in vitro cell migration and invasion abilities. Sphere formation and Hoechst 33342 exclusion assays were applied to examine the formation of sphenoid colonies and side population after silibinin treatment. Both intravesical orthotopic model and tail-vein injection metastasis model were generated to detect the effects of silibinin on in vivo tumor growth and distant metastasis. Western blot, quantitative PCR and luciferase reporter assays were used to analyze the regulation of GSK3β/β-catenin/ZEB1 signaling and the expression of its downstream molecular effectors (i.e., cytokeratins, vimentin, MMP2 and CD44) after silibinin treatment. Immunohistochemistry analysis was performed for evaluating p-GSK3β (Ser9), ZEB1, vimentin and CD44 expression in orthotopic xenograft tissues after silibinin treatment. RESULTS Silibinin treatment not only suppressed in vitro cell migration and invasion of metastatic T24-L cells, but also decreased bladder cancer lung metastasis and prolonged animal survival in vivo. Mechanistically, silibinin could inhibit GSK3β phosphorylation, β-catenin nuclear translocation and transactivation, and ZEB1 gene transcription that subsequently regulated the expression of cytokeratins, vimentin and MMP2 to reverse epithelial-mesenchymal transition (EMT). On the other hand, silibinin inhibited ZEB1 expression and then suppressed the properties of cancer stem cells (CSC), which were evidenced as decreased spheroid colonies formation, side population, and the expression of stem cell factor CD44. CONCLUSIONS Overall, this study reveals a novel effect and mechanism for silibinin targeting bladder cancer metastasis, in which inactivation of GSK3β/β-catenin/ZEB1 signaling by silibinin leads to the reversal of EMT phenotypes and CSC properties. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e225 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Kai-jie Wu More articles by this author Jian-cheng Zhou More articles by this author Zhong-yun Ning More articles by this author Jin Zeng More articles by this author Jin-hai Fan More articles by this author Da-lin He More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...