Survivin shRNA重组慢病毒构建及对A549细胞增殖的影响

BACKGROUND AND OBJECTIVE: Survivin is a member of the inhibitor of apoptosis family of proteins.The Survivin protein is highly expressed in most human tumors, but it is completely absent in terminally differentiated cells.Consequently, Survivin is an ideal target for cancer therapy because cancer cells are targeted and normal cells are left alone.The aim of this study is to construct a lentivirus-shRNA vector, and to disrupt the expression of Survivin in A549 cells.The effect of sh-RNA Survivin on A549 cells was analyzed. METHODS: Target DNA sequences of Survivin shRNA were designed to obtain recombinant plasmids.After the plasmids were transfected into 293T cells, the virus was collected.Hela cells were used to detect the virus titer.Survivin mRNA and protein expression in the infected A549 cells were detected via reverse transcription polymerase chain reaction and Western blot analysis.The proliferation of A549 cells were detected via 3-(4, 5-dimethylthiazolyl)-2, 5-diphenyltetrazolium bromide and flow cytometry assays. RESULTS: The recombinants were successfully constructed, and Survivin expression was inhibited.The cells were blocked at the G(2)/M phase. CONCLUSION: Recombinant lentivirus with shRNA targeting Survivin was successfully constructed.The lentivirus can down-regulate Survivin expression in A549 cells as well as inhibit proliferation, and is hence a potential gene therapy for lung cancer.