核糖核酸干扰沉默survivin基因对A549细胞增殖及顺铂敏感性的影响

目的：利用核糖核酸干扰（RNA interference,RNAi）技术沉默抗凋亡基因survivin,观察其对人肺腺癌细胞A549增殖以及顺铂药物敏感性的影响。方法：将靶向survivin的基因片段插入带有绿色荧光蛋白基因的载体后构建重组质粒,将其导入A549细胞,RT-PCR分析转染前后survivin mRNA的表达情况,免疫荧光及Western blot分析转染前后survivin蛋白的表达情况,TUNEL法检测细胞凋亡情况,MTT法检测转染后A549细胞对顺铂的敏感性变化。结果：成功构建pGenesil1。1-survivin重组质粒。与对照组相比,转染重组质粒后,survivin mRNA和蛋白的表达明显降低,抑制率分别为65%和71%;细胞凋亡率增加;转染前顺铂对A549细胞的IC50为转染后的7.68倍,两者比较,差异有统计学意义（P〈0.05）。结论：靶向survivin的RNA干扰表达载体能下调survivin基因表达,增强A549细胞对顺铂药物敏感性。

Silence of Survivin by RNA Interference Influence the Proliferation of A549 Cells and Sensitivity of Cisplatin

Objective:Inhibit expression of survivin with RNA intrference technique,to observe its effect on proliferation of A549 cells and sensitivity of A549 cells to cisplatin.Methods:The DNA sequence targeting human survivin was inserted into the plasmid which contain green fluorescent protein gene,and the recombinant plasmid were constructed. The recombinant plasmids were transfected into A549 cells.The levels of survivin mRNA and survivin protein were detected by RT-PCR and immunofluorescence staining and Wester...