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• W2604453411 abstract "You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III1 Apr 2017MP65-20 EPIGENETIC PRIMING WITH 5-AZACITIDINE INCREASES SENSITIVITY OF BLADDER CANCER CELLS TO CHEMOTHERAPEUTIC AGENTS Takahiro Yoshida, Anup Sharma, Max Kates, Niklai Sopko, Xiaopu Liu, Noah Hahn, David McConkey, Nita Ahuja, and Trinity Bivalacqua Takahiro YoshidaTakahiro Yoshida More articles by this author , Anup SharmaAnup Sharma More articles by this author , Max KatesMax Kates More articles by this author , Niklai SopkoNiklai Sopko More articles by this author , Xiaopu LiuXiaopu Liu More articles by this author , Noah HahnNoah Hahn More articles by this author , David McConkeyDavid McConkey More articles by this author , Nita AhujaNita Ahuja More articles by this author , and Trinity BivalacquaTrinity Bivalacqua More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2014AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bladder cancer is characterized by high prevalence of mutations in chromatin regulatory genes. Therefore, the epigenome presents multiple candidate targets for drug development. In this study, we assessed the effects of 5-azacitidine (AZA), which is a DNA methyltransferase inhibitor, on DNA methylation in luminal and basal bladder cancer cells. We also tested the priming effect of AZA on sensitivity of cells to cisplatin and gemcitabine METHODS Bladder cancer cell lines RT4 (luminal), 5637 (basal,″epithelial″), and J82 (basal, ″mesenchymal″/claudin-low) were treated with AZA in various conditions to determine optimal demethylation dosing schedules. Methylation levels were evaluated by pyrosequencing 11 CpG islands in long interspersed nucleotide elements-1 (LINE-1) repetitive element. To test priming effect of AZA, cell lines were pretreated with AZA at a dose that had no cytotoxic effects (500 and 1000 nM) for 5 days. Demethylation was confirmed and cells were subsequently subjected to cisplatin and gemcitabine treatment. Cell viability was evaluated using the CellTiter-Glo® assay. IC50's were calculated based on the results of the viability assays. RESULTS CpG islands in LINE-1 in the cell lines were demethylated by AZA treatment in a dose- and time-dependent manner. Although AZA suppressed proliferation of cell lines during treatment, it did not affect subsequent proliferation of cells after withdrawal of AZA. Pretreatment with AZA significantly decreased the IC50′s of cisplatin in the RT4 and 5637 cells from 2.82 to 0.78 and 0.67 to 0.37 μg/ml, respectively (Figure 1). The IC50's of gemcitabine were also significantly decreased in the RT4 and 5637 cells with pretreatment from 1.01 to 0.59 and 3.53 to 2.32 ng/ml, respectively (Figure 1). The IC50′s for cisplatin and gemcitabine did not change with AZA pretreatment in the J82 cell line. CONCLUSIONS AZA efficiently demethylated DNA in both luminal and basal bladder cancer cell lines. Pretreatment of AZA sensitized ″epithelial″ bladder cancer cells (RT4 and 5637) to cisplatin and gemcitabine. Combination of AZA epigenetic priming with chemotherapeutic agents warrants further investigation to delineate the mechanism of cytotoxic effect with priming in both NMIBC and MIBC. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e861 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Takahiro Yoshida More articles by this author Anup Sharma More articles by this author Max Kates More articles by this author Niklai Sopko More articles by this author Xiaopu Liu More articles by this author Noah Hahn More articles by this author David McConkey More articles by this author Nita Ahuja More articles by this author Trinity Bivalacqua More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ..." @default.
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• W2604453411 title "MP65-20 EPIGENETIC PRIMING WITH 5-AZACITIDINE INCREASES SENSITIVITY OF BLADDER CANCER CELLS TO CHEMOTHERAPEUTIC AGENTS" @default.
• W2604453411 doi "https://doi.org/10.1016/j.juro.2017.02.2014" @default.
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