【摘 要】
：
应用RNA干扰技术(RNAi)研究针对凋亡抑制因子survivin的siRNA抑制肝癌细胞株内源survivin基因的表达.转染重组质粒pshRNA-survivin至肝癌细胞株SMMC-7721,通过免疫荧光、蛋白
【机 构】
：
重庆医科大学病毒性肝炎研究所,The University of Chicago Medical Center
论文部分内容阅读
应用RNA干扰技术(RNAi)研究针对凋亡抑制因子survivin的siRNA抑制肝癌细胞株内源survivin基因的表达.转染重组质粒pshRNA-survivin至肝癌细胞株SMMC-7721,通过免疫荧光、蛋白质印迹和半定量RT-PCR检测survivin蛋白表达及mRNA转录水平的变化.结果表明:构建的三种重组质粒pshRNA-survivin1/2/3均能明显抑制survivin基因的表达;应用免疫荧光检测survivin基因的表达,转染重组质粒pshRNA-survivin的实验组survivin荧光强度明显低于转染载体pTZU6+1和pshRNA-GFP对照组;蛋白质印迹结果表明,重组质粒pshRNAsurvivin明显抑制survivin蛋白的表达,抑制率为62%～78%,通过半定量RT-PCR检测到survivin基因mRNA转录明显减少,抑制率为57%～64%.由上述结果可以得出结论:重组质粒pshRNA-survivin可明显抑制SMMC7721细胞内源survivin的表达和mRNA的转录,为survivin介导的肿瘤基因沉寂疗法提供实验基础.
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