应用siRNA技术下调survivin基因表达以增强SGC7901胃癌细胞对顺铂的敏感性  

Enhanced sensitivity to cisplatin in human gastric carcinoma cell line SGC7901 by the downregulation of the survivin gene with siRNA technology

作　　者：丁瑾[1] 孔庆兖[1] 

机构地区：[1]徐州医学院病理学教研室,江苏徐州221002

出　　处：《徐州医学院学报》2010年第5期305-308,共4页Acta Academiae Medicinae Xuzhou

基　　金：江苏省教育厅自然科学研究计划项目(04KJB310143)

摘　　要：目的利用小干扰RNA(siRNA)技术特异性下调survivin基因的表达,检测SGC7901胃癌细胞转染前后对顺铂敏感性的变化。方法实验分为空白对照组(control组)、单用顺铂组(CDDP组)、脂质体组(Lip组)、单用survivin siRNA组(siRNA组)、转染survivin siRNA(转染组)及转染survivin siRNA联合顺铂作用组(联合作用组)。人工合成survivin siRNA,通过Lip包裹将siRNA转染胃癌细胞SGC7901,荧光显微镜下观察转染情况。MTT法检测各组细胞的增殖抑制率,Western blot及免疫细胞化学法检测survivin蛋白的表达,Hoechst染色观察细胞凋亡的形态改变。结果Hoechst染色荧光显微镜下control组、Lip组及siRNA组细胞核呈正常的蓝色,而CDDP组、转染组及联合作用组细胞核染色增强、核质浓缩、核碎裂,呈凋亡改变;联合作用组细胞增殖抑制率(63.93±4.22)%明显高于其余各组(P<0.05);转染组及联合作用组survivin蛋白表达明显低于其他各组(P<0.05)。结论应用siRNA技术下调SGC7901胃癌细胞中survivin基因的表达,能够增加其对顺铂的药物敏感性。Objective To detect the variations in cisplatin sensitivity of human gastric carcinoma cell line SGC7901 before and after transfection to specifically downregulate the expression of surviving gene with small interfering RNA（siRNA） technology.Methods This experiment is divided into the control group,cisplatin group（CDDP group）,lipsome group（lip group）,siRNA group,transfection group and siRNA combined CDDP group（combined group）.Survivin siRNA was artificially synthesized and then transfected into gastric cancer cells with liposome.The transfection effects were determined by fluorescence microscopy.The inhibitory rate of cell proliferation was assayed by MTT,and the expression of survivin protein was determined by Western blot and immunocytochemical staining.The morphological changes of the apoptotic cells were observed by Hoechst staining.Results As observed under the fluorescence microscope after Hoechst staining,the nuclei presented as normal blue in the control group,lip group and siRNA group,while CDDP group,siRNA transfection group and siRNA combined cisplatin group had such changes as reinforced nuclear staining,karyopyknosis and karyorrhexis with apoptotic changes.The inhibition rate of cell proliferation in siRNA combined cisplatin group（63.93±4.22）% was markedly higher than the other groups（P0.05）.Compared with other groups,the expression of survivin protein in the transfection group and siRNA combined cisplatin group was significantly reduced,and the differences had statistical significance（P0.05）.Conclusion The application of siRNA technology to downregulate the survivin gene expression of human gastric carcinoma cell line SGC7901 is able to enhance its drug sensitivity to cisplatin.

关 键 词：SURVIVIN SIRNA 胃癌 顺铂 

分 类 号：R735.2[医药卫生—肿瘤]