中国肺癌杂志2011，Vol.14Issue(12)：903-907,5.DOI:10.3779/j.issn.1009-3419.2011.12.01
Survivin shRNA重组慢病毒构建及对A549细胞增殖的影响
Effect of shRNA Survivin-mediated Inhibition of Proliferation in A549 Cells
武丽红 1王金河 1王茜 1贾懂懂 1梁建琴1
作者信息
- 1. 100091北京,中国人民解放军第309医院结核病研究所
- 折叠
摘要
Abstract
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 AS49 cells. The effect of shRNA Survivin on AS49 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 G2/M phase. Conclusion Recombinant lentivirus with shRNA targeting Survivin was successfully constructed. The lentivirus can down-regulate Survivin expression in AS49 cells as well as inhibit proliferation, and is hence a potential gene therapy for lung cancer.关键词
慢病毒/Survivin/短发夹RNA/增殖Key words
Lentivirus/Survivin/shRNA/Proliferation分类
医药卫生引用本文复制引用
武丽红,王金河,王茜,贾懂懂,梁建琴..Survivin shRNA重组慢病毒构建及对A549细胞增殖的影响[J].中国肺癌杂志,2011,14(12):903-907,5.
