shRNA抑制人肺癌细胞A549 SURVIVIN基因表达

目的 构建存活素(SURVIVIN)基因的短发夹RNA(shRNA)表达质粒,为肺癌RNA干扰(RNAi)途径的基因治疗打下基础.方法 设计并合成SURVIVIN短发夹结构模板序列,与穿梭载体重组质粒后转染人肺癌细胞A549,通过RT-PCR法筛选出抑制效果最强的质粒;MTT和Western blot法检测该质粒对细胞增殖和SURVIVOIN表达的影响.结果 构建和筛选出抑制作用最强的质粒;该质粒能明显抑制A549细胞的增殖和SURVIVIN的表达.结论 在RNAi的介导下,SURVIVIN的表达被下调,为肺癌的基因治疗提供实验依据.

shRNA inhibits SURVIVIN expression in human lung cancer cell A549

Objective To construct shRNA expression vector of SURVIVIN for RNAi-mediated therapy of lung cancer.Methods DNA templates of SURVIVIN shRNA were designed,synthesized and cloned into the shuttle vector to get recombinant plasmids.After plasmids were transfected into A549 cells,the one with the most repression was screened by means of RT-PCR.Then MTT and Western blot were done to ascertain whether the proliferation of A549 cells were inhibited and SURVIVIN was down-regulated.Results The recombinants were constructed and screened successfully.The proliferation of A549 cells and SURVIVIN expression were repressed.Conclusion Mediated by RNAi,SURVIVIN expression was down-regulated,which suggested a potential gene therapy of huaman lung cancer.