CAJ | 학술논문

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sur-vivin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＝1620．26， 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＝2284．205，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。
목적：통과RNA간우억제식관암세포Eca-109 survivin기인적표체，관찰survivin기인침묵대Eca -109세포증식여조망적영향。방법：구건파향survivin기인적siRNA표체재체병차조지질체은정전염Eca -109세포（간우조），동시설전염무관간우질립적Eca-109세포위음성대조，미전염세포위공백대조。채용Western blot 검측sur-vivin기인침묵효과，MTT、평판극륭형성실험급류식세포술분석간우전후Eca-109세포증식여조망적변화。결과：여량대조조상비，간우조Survivin단백적표체강저（F＝79．397，P＜0．001）；MTT검측결과현시，간우조억제세포증식효과우우량대조조，이평판극륭형성실험현시간우조극륭형성솔저우량대조조（F＝1620．26， P＜0．001）；류식세포술분석현시，간우조조망지수고우량대조조（F＝2284．205，P＜0．001）。결론：RNA간우가특이성침묵survivin기인적표체，억제Eca-109세포적증식，유도세포조망。
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.