Abstract: To construct expression vector with short hairpin RNA of survivin，which paves the way for lung cancer RNAi-mediated therapy. Methods Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized.After annealed,the double Objective To construct shRNA expression vector of SURVIVIN for RNAi-mediated therapy on 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 theoretically based on gene therapy on huaman lung cancer.