沉默zeste基因增强子同源物2对结肠癌HT29细胞奥沙利铂化疗敏感性的影响  

Effects of silencing enhancer of zeste homolog 2 on oxaliplatin chemosensitivity of colon cancer HT29 cells

作　　者：吴万庆[1] 傅聿铭[1] 蒋营浩 郭晓磊[1] 闫军浩 Wu Wanqing;Fu Yuming;Jiang Yinghao;Guo Xiaolei;Yan Junhao(Department of General Surgery,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第五附属医院普外科,郑州450052

出　　处：《中华实验外科杂志》2024年第8期1747-1750,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20220562)。

摘　　要：目的探讨zeste基因增强子同源物2(EZH2)对结肠癌HT29细胞奥沙利铂化疗敏感性的影响及机制。方法实时定量聚合酶链反应(Real-time PCR)法检测EZH2小干扰RNA(siRNA)转染HT29细胞24 h后转染组(转染EZH2 siRNA)、空白对照组、阴性对照组(转染Control siRNA)mRNA相对表达量,蛋白质印迹法(Western blot)法检测EZH2蛋白相对表达量;使用不同浓度的奥沙利铂处理转染24 h的细胞,48 h后噻唑蓝(MTT)法检测细胞存活率,计算半数抑制浓度(IC50);使用50μg/μl的奥沙利铂处理转染24 h的细胞,48 h后流式细胞仪检测细胞凋亡,Western blot法检测Survivin、环氧合酶-2(COX-2)蛋白相对表达量。组间采用用单因素方差分析和LSD-t检验。结果转染组EZH2 mRNA低于空白对照组及Control siRNA组(105.1±24.3比545.9±88.6、558.4±60.4,F=496.2,P<0.05);转染组EZH2蛋白相对表达量低于空白对照组及Control siRNA组(75.4±10.9比395.8±29.7、391.7±41.7,F=1109.6,P<0.05)。不同浓度(0、6.25、12.5、25、50、100μg/μl)奥沙利铂处理后转染组细胞存活率低于空白对照组及Control siRNA组,转染组IC50低于空白对照组及Control siRNA组(26.94μg/μl比63.24、56.74μg/μl)。50μg/μl奥沙利铂处理后转染组细胞凋亡率高于空白对照组及Control siRNA组(45.4±4.9比7.1±1.6、6.8±2.0,F=95.5,P<0.05)。转染组Survivin、COX-2蛋白相对表达量均低于空白对照组及Control siRNA组(43.2±8.8、47.0±10.3比252.5±31.9、248.5±23.6,F=790.2,P<0.05;43.2±8.8、47.0±10.3比246.0±7.0、264.8±26.0,F=1558.1,P<0.05)。结论利用siRNA沉默EZH2表达可通过促进细胞凋亡提高结肠癌HT29细胞对奥沙利铂的化疗敏感性。ObjectiveTo investigate the effects of enhancer of zeste homolog 2(EZH2)on the chemosensitivity of colon cancer HT29 cells treated by oxaliplatin and mechanisms.MethodsReal-time quantitative polymerase chain reaction(Real-time PCR)was used to detect the relative expression levels of mRNA in the transfection group[transfected with EZH2 small interfering RNA(siRNA)],blank control group(transfected with Control siRNA),and negative control group after 24 h of transfection of HT29 cells with EZH2 siRNA,and the relative expression of EZH2 protein was detected by Western blotting.Different concentrations of oxaliplatin were used to treat transfected cells for 24 h,and the cell viability was detected by methyl thiazolyl tetrazolium(MTT)assay at 48 h after treatment.The half-maximal inhibitory concentration(IC 50)was calculated.The cells transfected for 24 h were treated with 50μg/μL of oxaliplatin,and cell apoptosis was detected by flow cytometry at 48 h after treatment.The relative expression levels of Survivin and cyclooxygenase-2(COX-2)proteins were detected by Western blotting.One-way ANOVA and LSD-t test were used to compare data between groups.ResultsThe relative expression level of EZH2 mRNA in EZH2 siRNA group was lower than that in control group and control siRNA group(105.1±24.3 vs.545.9±88.6,558.4±60.4,F=496.2,P<0.05).The relative expression of EZH2 protein in EZH2 siRNA group was lower than that in blank control group and control siRNA group(75.4±10.9 vs.395.8±29.7,391.7±41.7,F=1109.6,P<0.05).After oxaliplatin treatment with different concentrations(0,6.25,12.5,25,50,100μg/μL),the cell survival rate in EZH2 siRNA group was lower than that in blank control group and control siRNA group,and the IC 50 of EZH2 siRNA group was lower than that in blank control group and control siRNA group(26.94μg/μL vs.63.24,56.74μg/μL).The apoptosis rate of 50μg/μL oxaliplatin treated in EZH2 siRNA group was lower than that in control group and control siRNA group(45.4±4.9 vs.7.1±1.6,6.8±2.0,F=95.5,P<0.05).The

关 键 词：结肠癌 奥沙利铂 小干扰RNA 

分 类 号：R735.35[医药卫生—肿瘤]