生存素
小发夹RNA
分子生物学
转染
A549电池
生物
细胞凋亡
癌症研究
癌细胞
免疫印迹
化学
细胞培养
基因
癌症
生物化学
遗传学
基因敲除
作者
Lihong Wu,Jinhe Wang,Qian Wang,Dong-Dong Jia,Juhua Liang
出处
期刊:PubMed
日期:2011-12-01
标识
DOI:10.3779/j.issn.1009-3419.2011.12.01
摘要
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.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.The recombinants were successfully constructed, and Survivin expression was inhibited. The cells were blocked at the G2/M phase.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.
科研通智能强力驱动
Strongly Powered by AbleSci AI