曲古霉素A对人膀胱癌细胞EJ增殖的影响及其作用机制  

Effect of trichostatin A on the proliferation of human bladder cancer cell line EJ and its underlying mechanism

作　　者：冷俊 周晔 施利琴 张莉 LENG Jun;ZHOU Ye;SHI Liqin;ZHANG Li(Department of Clinical Laboratory,Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Chongming Branch,Shanghai 202150,China)

机构地区：[1]上海交通大学医学院附属新华医院崇明分院检验科,上海2021500

出　　处：《癌症进展》2021年第5期454-457,共4页Oncology Progress

基　　金：上海市科学技术委员会科研计划项目(17DZ1930309)。

摘　　要：目的探讨曲古霉素A(TSA)对人膀胱癌细胞EJ增殖及WNT抑制因子-1(WIF-1)mRNA表达的影响,并分析其可能的作用机制。方法采用四甲基偶氮唑蓝(MTT)比色法检测不同浓度的TSA对人膀胱癌细胞EJ增殖的影响;采用流式细胞术检测经TSA处理后人膀胱癌细胞EJ的细胞周期分布情况;采用实时逆转录聚合酶链反应(RT-PCR)检测用药前后人膀胱癌细胞EJ中WIF-1 mRNA的表达情况;采用染色质免疫沉淀技术分析用药前后WIF-1基因启动子区组蛋白H3第9位赖氨酸(H3-K9)的乙酰化状态。结果MTT比色法检测结果显示,处理72 h后,0.1、0.2、0.4、0.8和1.6μmol/L人膀胱癌细胞EJ的增殖抑制率均高于空白对照组(P﹤0.05)。流式细胞仪检测结果显示,人膀胱癌细胞EJ经0.2、0.4和0.8μmol/L浓度的TSA处理72 h后,G0/G1期、G2/M期细胞的数量随着TSA浓度的升高而逐渐增加,S期细胞的数量随着TSA浓度的升高而逐渐减少。RT-PCR检测结果显示,与空白对照组相比,不同浓度(0.2、0.4和0.8μmol/L)的TSA处理24、48、72 h后,人膀胱癌细胞EJ中WIF-1 mRNA的相对表达量均升高(P﹤0.05)。琼脂糖凝胶电泳结果显示,电泳条带符合预期的片段长度,经TSA处理后,人膀胱癌细胞EJ中扩增出目的片段,且随着TSA浓度的增加,目的片段的产物增多。结论TSA对人膀胱癌细胞EJ的增殖具有抑制作用,其作用机制可能涉及细胞周期阻滞、WIF-1基因启动子区组蛋白H3-K9乙酰化水平升高及该基因的表达水平升高。Objective To investigate the effect of trichostatin A(TSA)on the expression of WNT inhibitory factor-1(WIF-1)gene in EJ cell,a human bladder cancer cell line,and their proliferation,and to further explore the possible underlying mechanism.Method The effect of different concentrations of TSA on the proliferation of human bladder cancer cell EJ was detected by methyl thiazolyl terazolium(MTT)assay.The cell cycle distribution of bladder cancer cell EJ after TSA treatment was detected by flow cytometry.Real-time reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of WIF-1 mRNA in human bladder cancer cell EJ before and after medication.The acetylation status of histone H3 lysine 9(H3-K9)in the promoter region of WIF-1 gene was measured by chromatin immunoprecipitation.Result MTT colorimetric test showed that the inhibitory rate of EJ cell treated with 0.2,0.4,0.8 and 1.6μmol/L TSA for 72 h was higher than that of the blank control group(P<0.05).Flow cytometry indicated that the cell number in G0/G1 and G2/M phase increased with the increase of TSA concentration while cells in S phase decreased with the adding concentration.The RT-PCR results presented that the relative expression of WIF-1 mRNA in bladder cancer cell EJ after 24,48,72 h treatment with different concentrations(0.2,0.4 and 0.8μmol/L)of TSA treatment increased compared with that of the blank control group(P<0.05).The results of agarose gel electrophoresis showed that the electrophoresis band corresponded with the expected fragment length.After TSA treatment,the target fragment was amplified in the bladder cancer cell EJ,and the product of the target fragment increased with the increase of TSA concentration.Conclusion TSA has an inhibitory effect on the proliferation of bladder cancer cell EJ,and its underlying mechanism may involve cell cycle arrest,up-regulation of histone H3-K9 acetylation level in the promoter region of WIF-1 gene and the increase of the expression level of this parental gene.

关 键 词：曲古霉素A 膀胱癌 WNT抑制因子1 组蛋白乙酰化 增殖 

分 类 号：R737.14[医药卫生—肿瘤]