RNA干扰沉默survivin基因对Eca-109细胞增殖与凋亡的影响

Effects of silencing survivin gene by RNA interference on proliferation and apoptosis of Eca-109 cells

目的:通过RNA干扰抑制食管癌细胞Eca-109 survivin基因的表达,观察survivin基因沉默对Eca-109细胞增殖与凋亡的影响。方法:构建靶向survivin基因的siRNA表达载体并借助脂质体稳定转染Eca-109细胞(干扰组),同时设转染无关干扰质粒的Eca-109细胞为阴性对照,未转染细胞为空白对照。采用Western blot检测survivin基因沉默效果,MTT、平板克隆形成实验及流式细胞术分析干扰前后Eca-109细胞增殖与凋亡的变化。结果:与两对照组相比,干扰组Survivin蛋白的表达降低(F=79.397,P<0.001);MTT检测结果显示,干扰组抑制细胞增殖效果优于两对照组,而平板克隆形成实验显示干扰组克隆形成率低于两对照组(F=162.026,P<0.001);流式细胞术分析显示,干扰组凋亡指数高于两对照组(F=2 284.205,P<0.001)。结论:RNA干扰可特异性沉默survivin基因的表达,抑制Eca-109细胞的增殖,诱导细胞凋亡。

Aim：To inhibit endogenous survivin expression by RNA interference technology and investigate the effects on proliferation and apoptosis of Eca-109 cells.Methods： An siRNA expressing vector targeting survivin was constructed and transfected into Eca-109 cells via Lipofectamine TM 2000 to establish stable transfection cell line .The Eca-109 cells transfected non-sense sequence and those untransfected were used as control .The expression level of Survivin protein was detected by Western blot to observe the interference effect;cell proliferation of Eca-109 cells was determined by MTT assay and colony formation assay;cell apoptosis of Eca -109 cells was measured by flow cytometry analysis .Results： Compared with the 2 control groups , the expression level of Survivin protein in the interference group was significantly reduced （ F=79.397,P〈0.001）;cell proliferation in the interference group was distinctly inhibited （F=162.026,P〈00.01 ）;the ap-optosis index in the interference group was significantly increased （F=2 284.205,P〈0.001）.Conclus ion： Knockdown of endogenous survivin via specific RNA interference can effectively inhibit cell proliferation and significantly induce cell apoptosis of Eca-109 cells.