Induction of cell cycle arrest in bladder cancer BIU-87 cells by trichostatin A and bacillus calmette-guerin

Abstract

Objective To investigate the influence of trichostatin A (TSA) and bacillus calmetteguerin (BCG) on cell cycle of human bladder cancer BIU-87 cells and the expression of related genes,and to explore the involved mechanism.Methods After treatment,cell growth was measured by methyl thiazol tetrazolium (MTT) assay.The cell cycle distribution was examined by flow cytometry.The expression of p27kip1 and Cyclin D1 mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR).The expressional level of p27kip1 and Cyclin D1 protein was detected by Western blotting.Results TSA and BCG significantly inhibited the proliferation of BIU-87 cells in a time-and dose-dependent manner.Moderate or high doses of TSA combined with BCG exerted the synergic effect.After BIU-87 cells were treated with TSA and BCG,cells were blocked at G0/G1 phase.The cell proportion of G0/G1 phase in the control was (44.39 ± 3.42) ％,however,it was (78.3 ± 4.12) ％ at 24 h in TSA + BCG-treated group (P ＜ 0.05),accompanied by increased p27kip1 mRNA expression and decreased Cyclin D1 mRNA expression.The results of Western blotting showed that the p27kip1 protein level was promoted and the Cyclin D1 protein level dropped after TSA and BCG treatment.Conclusion The cells were blocked at G0/G1 phase with upregulation of the expression of p27kip1 and down-regulation of the expression of Cyclin D1,suggesting TSA and BCG can inhibit bladder cancer cells growth in vitro through inducing cell cycle arrest,which might be related to the expression of p27kip1 and Cyclin D1. Key words: Bladder cancer; Trichostatin A; Bacillus calmette-guerin; Cell cycle