摘要
目的 观察下调激素受体相关受体α（ERRα）表达后奥沙利铂处理结肠癌细胞株增殖及凋亡变化并探讨其机制.方法 结肠癌细胞株 Colo-205、HCT-116、SW620及HT-29采用贴壁细胞法进行培养,按照给予的干预方式分为在奥沙利铂处理后分别给予ERRα抑制剂XCT790的XCT790-OHP-HCT-116组、给予siERRα转染HCT-116细胞下调ERRα表达的siERRα-OHP-HCT-116组及奥沙利铂干预组（OHP-HCT-116组）,对照组为未给予干预的无干预组（NC 组）.试验另分为给予 siERRα 转染HCT-116细胞下调ERRα表达的siERRα组和以siNegative Control进行转染的阴性对照组（siNC组）.以Western blot法和实时定量（qRT）-PCR法检测结肠癌细胞ERRα蛋白和mRNA水平表达,以ERRα抑制剂XCT790及siERRα下调ERRα表达,流式细胞术及甲基噻唑基四唑法检测结肠癌细胞凋亡及增殖.Western blot法、qRT-PCR检测细胞凋亡及增殖相关基因蛋白及 mRNA 表达.结果 ERRα 蛋白及mRNA在HCT-116中高于Colo-205、SW620及HT-29细胞株（P均〈0.05）;XCT790-OHP-HCT-116组细胞早期凋亡率高于NC组及OHP-HCT-116组（P均〈0.05）,XCT790-OHP-HCT-116组细胞培养72、96 h存活率低于NC组及OHP-HCT-116组（P均〈0.05）.采用siERRα转染HCT-116细胞下调ERRα表达后, siERRα-OHP-HCT-116组早期凋亡率低于 NC 组及 OHP-HCT-116组（P 均〈0.05）,siERRα-OHP-HCT-116组细胞培养72、96 h后存活率低于NC组及OHP-HCT-116组（P均〈0.05）;给予siERRα转染HCT-116细胞,与 siNC 组对比,siERRα 组 YAP1、p73、p63、MDM2、Capase 8、Capase 9蛋白水平下调（P 均〈0.01）,mRNA水平比较差异无统计学意义（P〉0.05）.结论 下调ERRα表达促进结肠癌细胞凋亡抑制增殖,增强奥沙利铂对结肠癌细胞的杀伤作用.
Objective To observe the changes of proliferation and apoptosis of colon cancer cell line treated with oxaliplatin after the downregulation of ERRα and to investigate the mechanism.Methods Colon cancer cell lines Colo-205,HCT-116,SW620 and HT-29 were cultured by adherent cells and in accordance with the given intervention,they were divided into group XCT790-OHP-HCT-116（after oxaliplatin treatment,ERR αinhibitor XCT790 was administered）,group siERRα-OHP-HCT-116（after oxaliplatin treatment,siERR α was transfected into HCT-116 cells and downregulated ERR αexpression）,oxaliplatin intervention group（group OHP-HCT-116）and the control group（NC group）which was given no intervention.The experiment was divided into siERR αgroup with siERR α transfected with HCT-116 cells,downregulated ERR αexpression and the negative control group（siNC group）transfected with siNegative Control.Using Western blot method and real-time quantitative（qRT）-PCR for the detection of colorectal cancer cell ERRαprotein and mRNA expression,the expression of ERR αwas downregulated by ERR αinhibitors XCT790 and siERR,and apoptosis and proliferation of colon cancer cells were detected by flow cytometry and MTT.Western blot and qRT-PCR were used to detect apoptosis and proliferation-related gene proteins and mRNA expression.Results ERR αand mRNA protein in HCT-116 were higher than those of Colo-205,SW620 and HT-29 cell lines（P〈0.05）; in the XCT790-OHP-HCT-116 group,the early apoptosis rate was higher than those of the NC group and OHP-HCT-116 group（P〈0.05）,the survival rate of cell culture in 72 and 96 h in the XCT790-OHP-HCT-116 group was lower than those in the NC group and OHP-HCT-116 group（P〈0.05）.The siERR α HCT-116 cells transfected with down-regulation of ERR expression,siERR α -OHP-HCT-116 group early apoptosis rate was lower than those of NC group and OHP-HCT-116 group（P〈0.05）,siERR -OHP-HCT-116 group cells cultured for 72 and 96 h after the survival rate was lower than the NC group and OHP-HCT-116 group（P〈0.05）;After the downregulation of ERRαby siERR alpha transfected with HCT-116 cells,the early apoptotic rate in the group siERRα-OHP-HCT-116 was lower than that in the group NC and group OHP-HCT-116（P〈0.05）,the survival rate of the group siERRα-OHP-HCT-116 after 72 and 96 h were lower than those in the group NC and group OHP-HCT-116（P〈0.05）,siERR α was transfected into HCT-116 cells,compared with the siNC group,YAP1,p73,p63,MDM2, Capase 8,Capase 9 protein in the siERR group decreased（P〈0.01）,there was no significant difference in the level of mRNA（P〉0.05）.Conclusion The downregulation the expression of ERRαcan promote colon cancer cell apoptosis,inhibit proliferation,and enhance the killing effect of oxaliplatin on colon cancer cells.
出处
《中国综合临床》
2017年第10期865-869,共5页
Clinical Medicine of China
基金
国家自然科学基金资助项目（81672427）
辽宁省博士科研启动基金（201601417）
作者简介
通信作者：刘放，Email：liufang@cancerhosp-In-cmu．com
