曲古抑菌素A和卡介苗诱导膀胱癌BIU-87细胞周期阻滞的研究  

Induction of cell cycle arrest in bladder cancer BIU-87 ceils by trichostatin A and bacillus calmetteguerin

作　　者：田雨冬[1] 潘铁军[2] 叶章群[3] 李功成[2] 

机构地区：[1]郑州大学第一附属医院泌尿外科,450052 [2]广州军区武汉总医院泌尿外科 [3]华中科技大学同济医学院附属同济医院泌尿外科

出　　处：《中华实验外科杂志》2014年第4期794-796,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省科技攻关计划资助项目（2007AA301B15-4）

摘　　要：目的观察曲古抑菌素A（TSA）和卡介苗（BCG）及两者联合对人膀胱癌BIU-87细胞周期及相关基因表达的影响，并探讨其作用机制。方法采用噻唑蓝（MTT）法检测细胞生长抑制率，流式细胞术检测细胞周期分布的变化，逆转录-聚合酶链反应（RT-PCR）检测细胞p27kip1、细胞周期素（Cyclin）D1mRNA表达的变化，Westernblot检测细胞p27kipl、CyclinD1蛋白水平的变化。结果TSA和BCG及两者联合应用能明显抑制BIU-87细胞生长，呈时间和剂量依赖，相对中高剂量两者联用表现为协同作用。TSA和BCG能诱导BIU-87细胞G0/G1期阻滞，对照组G0/G1期细胞比例为（44．39±3．42）％，而TSA联合BCG组GO/G．期细胞比例达（78．3±4．12）％（P〈0．05）。RT-PCR结果示TSA和BCG及两者联用能显著诱导p27kpilmRNA的表达和显著抑制CyclinDImRNA的表达，两者联合与单用比较差异有统计学意义（P〈0．05）；Westernblot进一步在蛋白水平证实了上述结果。结论CyclinD1的表达下降和p27kipl的表达上调共同使细胞阻滞于G0/G1期，这表明TSA和BCG可通过诱导细胞周期阻滞而发挥体外抗膀胱癌作用，其作用机制可能涉及相关基因p27kipl、CyclinD1表达的调控。Objective To investigate the influence of trichostatin A （TSA） and bacillus calmette guerin （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 1327kipl and Cyclin D1 mRNA was assessed by reverse transcription-polymerase chain reaction （RT-PCR）. The expressional level of p27klvl 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. Moder ate 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 24h in TSA＋BCG-treated group （P〈 0. 05）, accompanied by increased p27kipl mRNA expression and decreased Cyclin D1 mRNA expression. The results of Western blotting showed that the p27k@ 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 up regulation of the expression of 1327klpl and down-regulation of the expression of Cyclin D1, suggesting TSA and BCG can inhibit bladder cancer ceils growth in vitro through inducing cell cycle arrest, which might be related to the expression of p27kipl and Cyclin D1.

关 键 词：膀胱癌 曲古抑菌素A 卡介苗 细胞周期 

分 类 号：R737.14[医药卫生—肿瘤]