siRNA下调Ets2对肾癌786-0细胞增殖的影响

目的:探讨E26转录因子2(E-twenty six2,Ets2)在肾癌细胞中的表达及其对肾癌细胞786-0增殖的影响及机制.方法:Westem Blot检测各肾癌细胞内Ets2的表达,并合成特异性siRNA下调肾癌细胞系786-0中Ets2的表达,利用MTT、平板克隆形成实验及流式细胞术检测其增殖能力的变化,通过Western Blot检测细胞周期相关分子表达水平变化.结果:Ets2在肾癌细胞内的表达比正常肾小管上皮细胞明显增高,siRNA能有效下调肾癌786-0细胞系中Ets2的表达.下调Ets2后786-0细胞增殖能力及克隆形成能力减弱(P＜0.01);细胞周期阻滞于G0/G1期;Ets2-siRNA介导的Ets2基因沉默下调了786-0细胞内CDK4与CyclinD1的表达,而上调了p21的表达.结论:Ets2可通过调节CDK4、CyclinD1及p21的表达影响肾癌细胞786-0的增殖能力.

Influence of siRNA targeting Ets2 gene on proliferation of renal cell carcinoma cell 786-0

Objective:To test the expression of E-twenty six2 (Ets2) in different renal cell carcinoma(RCC) cells and investigate the mechanism of Ets2 regulate 786-0 cells' proliferation.Methods:The expression of Ets2 in different RCC cells were tested by Western Blot.Design and synthesize the specific siRNA could down-regulate the expression of Ets2 in 786-0 cells.siRNA was transfected into 786-0 cells and the effect was detected by Western Blot.Proliferation were tested by the MTT assay,colony formation assay and flow cytometry.The expression levels of CDK4,CyclinD1 and p21 were detected by Western methods.Results:The expression of Ets2 in HK-2 cells was lower than that in RCC cells.siRNA can effectively down-regulate the expression of Ets2 in 786-0 cells.Down-regulation of Ets2 expression can inhibit the cell proliferation capacity.In addition,the expressions of CDK4 and CyclinD1 were decreased,while the expression of p21 was increased in siRNA groups.Conclusion:Ets2 might regulate CDK4-CyclinD1-p21 to promote proliferation in 786-0 cells.