shRNA inhibits SURVIVIN expression in human lung cancer cell A549
Liang Nian-ci
Abstract
Liang Nian-ci
Abstract
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.
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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.
Key concepts: Survivin, Small hairpin RNA, A549 cell, Transfection, RNA interference, Expression vector, Molecular biology, Cancer research