生存素siRNA表达质粒对MGC-803细胞增殖的影响  

Effects of survivin siRNA expression plasmid on proliferation of MGC-803 cells

作　　者：赵伟红[1] 郭俊明[1] 肖丙秀[1] 管忠 肖东升[1] 

机构地区：[1]宁波大学医学院,浙江省宁波市315211

出　　处：《世界华人消化杂志》2005年第19期2302-2305,共4页World Chinese Journal of Digestology

基　　金：浙江省医药卫生科学研究基金资助项目;No.2004A079;宁波市青年博士基金资助项目;No.2004A620007

摘　　要：目的:应用siRNA表达载体介导的RNAi技术,观察 RNA干扰沉默生存素基因对胃癌MGC-803细胞增殖的影响．方法:用脂质体介导将生存素siRNA表达质粒转染 MGC-803细胞,通过倒置显微镜观察转染后细胞形态学的变化,四甲基偶氮唑蓝(MTT)法检测细胞增殖活性,流式细胞术分析细胞周期的变化,逆转录聚合酶链式反应(RT-PCR)方法检测生存素mRNA的表达水平．结果:生存素siRNA表达质粒转染组MGC-803细胞变圆、浮起,生存素siRNA表达质粒明显下调MGC-803 细胞内生存素mRNA的表达,与空白对照组相比降低了48．2％,阻断细胞周期在G1期(77．4％),显著抑制细胞增殖,siRNA转染组细胞吸光度比空白组显著降低 (24 h:0．272±0．048 vs 0．576±0．018;48 h:0．270±0．060 vs 0．809±0．027;72 h:0．143±0．046 vs 1．015±0．075;均 P<0．01)．转染24,48和72 h后的抑制率分别为53．4％, 66．7％和86.3％．结论:应用siRNA表达载体介导的RNAi技术,可有效下调生存素在MGC-803细胞中的表达,并在体外抑制细胞的增殖．AIM： To silence the expression of survivin gene in MGC-803 cells by the siRNA expression vector-based RNA interference （RNAi） technique, and to investigate its effects on the proliferation of MGC-803 cells. METHODS： The survivin siRNA expression plasmid was transfected into MGC-803 cells by lipofectamine. Morphological changes of the cells were observed under invert microscope. The expression of survivin mRNA was determined by reverse transcription- polymerase chain reaction （RT-PCR）. The changes of cell cycle and the cell proliferation were analyzed by flow cytometry and MTT assay, respectively. RESULTS： Abnormal morphological changes of MGC-803 cells were observed in the group transfected with the survivin siRNA expression plasmid. The survivin siRNA expression plasmid significantly down-regulated the expression of survivin mRNA in MGC-803 cells with a percentage of 48.2% （ vs empty controls）, and itarrested the cell cycle in G1 phase （77.4%）. The cell proliferation was significantly inhibited, and the optical density in siRNA-transfected cells was markedly lower than that in the empty controls （24 h： 0.272 ± 0.048 vs 0.576 ± 0.018; 48 h： 0.270 ± 0.060 vs 0.809 ± 0.027; 72 h： 0.143 ± 0.046 vs 1.015 ± 0.075; all P 〈0.01）. The growth inhibitory rates of MGC-803 cells were 53.4%, 66.7%, and 86.3% after 24, 48, and 72 h of the transfection, respectively. CONCLUSION： The expression of survivin in MGC-803 cells can be down-regulated by the plasmid-based RNAi technique, and the down-regulation can inhibit the cell proliferation in vitro.

关 键 词：生存素 RNA干扰 细胞周期 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤] R587.1[医药卫生—临床医学]