MP39-17 MECP2 SILENCING IN HIGH GRADE BLADDER CANCER DERIVED HTB-5 CELL LINE AFFECTS CELLULAR PROLIFERATION AND MIGRATORY ABILITY

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research IV1 Apr 2014MP39-17 MECP2 SILENCING IN HIGH GRADE BLADDER CANCER DERIVED HTB-5 CELL LINE AFFECTS CELLULAR PROLIFERATION AND MIGRATORY ABILITY Shumaila Bilgrami, Zulfiqar Naqvi, Khalid Shaikh, El Nasir Lalani, and Farhat Abbas Shumaila BilgramiShumaila Bilgrami More articles by this author , Zulfiqar NaqviZulfiqar Naqvi More articles by this author , Khalid ShaikhKhalid Shaikh More articles by this author , El Nasir LalaniEl Nasir Lalani More articles by this author , and Farhat AbbasFarhat Abbas More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.1332AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES DNA methylation patterns and chromatin structure is significantly altered in many human neoplasms including bladder cancer. Aberrant promoter hypermethylation of tumor suppressor genes is now a well established mechanism of gene inactivation. Methyl binding proteins are believed to act as interpreters of DNA methylation signal. The methyl binding protein MeCP2 has been shown to bind specifically to the 5mCpGs that are flanked by A/T residues. (Klose R et al., 2005) and mediates transcriptional repression by recruiting histone deacetylases (Adrian Bird at al.,1998) and thus, serving as a bridge between two major regulatory mechanisms of gene expression-DNA methylation and histone deacetylation. METHODS A high grade bladder cancer cell line HTB-5 (ATCC) was transfected with psiRNA-hMeCP2 and psiRNA-LucGL3 (Invivogen), after testing for Mycoplasma contamination. CELL PROLIFERATION ASSAY Cellular proliferation was measured using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide]assay kit (ATCC)according to manufacturer’s protocol. CELL MIGRATION ASSAY Confluent monolayers of HTB-5 cells in 6-well plates were serum starved and incubated in minimum essential medium for 24hrs. A bee line (1mm thick) was created with a 10μl pipette tip. Cell migration was monitored at 24hr interval and photographed. RESULTS The results of proliferation assay showed that cell growth was drastically compromised in the MeCP2 silenced cells as compared to the mock transfected and untransfected HTB-5 cells after 24, 48, 72 and 96 hours of incubation (Fig.1) The migratory potential of MeCP2-silenced cells was evaluated by the cell migration/ wound healing assay. The untransfected and mock transfected HTB-5 cells were able to trespass the trough after 48 hours. On the other hand, MeCP2 silenced cells migrated slower than the control transfected and parental HTB-5 cells (Fig.2). CONCLUSIONS The results of our study demonstrated that MeCP2 silencing in high grade bladder cancer derived HTB-5 cells resulted in reduced proliferation and migratory ability. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e432 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Shumaila Bilgrami More articles by this author Zulfiqar Naqvi More articles by this author Khalid Shaikh More articles by this author El Nasir Lalani More articles by this author Farhat Abbas More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...