Survivin siRNA对肝癌细胞BEL-7402的增殖与对Survivin基因及蛋白表达的作用

目的:探讨小干扰RNA(siRNA)对BEL-7402细胞的生长抑制与生存素(Survivin)基因及蛋白表达的影响。方法:合成Survivin siRNA,以不同浓度转染BEL-7402细胞,MTT法测定细胞生长抑制率,RT-PCR、Western-Blot检测BEL-7402细胞Survivin mRNA及蛋白的表达差异。结果:BEL-7402细胞转染FAM荧光标记的siRNA后,细胞内有绿色荧光表达。分别以10、30、50nmol/LSurvivin siRNA转染细胞后,MTT测得的细胞生长抑制率分别为(38.80±1.71)%,(54.50±1.16)%,(66.79±1.10)%,RT-PCR测得的Survivin mRNA表达分别为0.42±0.02,0.28±0.01,0.12±0.01,Western-Blot测得Survivin蛋白表达分别为0.55±0.04,0.32±0.02,0.21±0.02,三项指标与空白对照组、阴性对照组、脂质体对照组比较有极显著性差异(P<0.01)。随着Survivin siRNA浓度的增加,细胞抑制率呈上升趋势,Survivin mRNA及Survivin蛋白的表达逐渐降低,呈浓度-效应关系。结论:Survivin siRNA能抑制BEL-7402细胞的增殖与Survivin基因和蛋白表达。

Effect of Survivin siRNA on cell proliferation and the expression of Survivin gene and protein in cell BEL-7402

Objective：To investigate the effects of siRNA on cell proliferation and the expression of Survivin gene and protein in cell BEL 7402. Methods：The siRNA targeting Survivin was synthesized and transfeeted into BEL-7402 in different concentrations. The killing effects on the cells were evaluated by MTT. The expression of Survivin mRNA gene was detected by RT-PCR and the expression level of Survivin protein was detected by western-blot. Results ：The green fluoreseet expression were observed in the infected BEL-7402. After the BEL-7402 cell was transfected with 10,30,50nmol/L dose Survivin siRNA respcetively,the rates of cells inhibition measured by MTT were （38.80± 1.71） %, （54.50± 1.16） %, （66.79± 1.10）%, the Survivin/ GAPDH gray value of Survivin mRNA by PT-PCR were 0.42±0.02, 0.28±0.01, 0.12±0.01, the survivin/β- actin of Survivin protein were 0. 55 ± 0. 04, 0. 32 ± 0. 02, 0. 21± 0. 02, the three indexes were significant different from control group, negative control group ,and lipofectamine control （P〈0.01）, with the increase of siRNA concentrations, the BEL 7402 inhibition rate was increased in different groups, the expressions of Survivin protein and mRNA were decreased, and there was a concentration-effect relationship. Conclusion：The Survivin siRNA can inhibite the expression of Survivin gene and cell proliferation of BEL7402.