RNA interference by expression of short hairpin RNAs suppresses bcl-xL gene expression in nasopharyngeal carcinoma cells

被引:8
作者
Liu, F
He, CW
Zhang, YF
Zhou, KY [1 ]
机构
[1] First Affiliated Hosp, Guangdong Med Coll, Inst Biochem & Mol Biol, Zhanjiang 524023, Peoples R China
[2] First Affiliated Hosp, Guangdong Med Coll, Dept Otorhinolaryngol, Zhanjiang 524023, Peoples R China
关键词
RNAi; plasmid; gene tranfection; short hairpin RNA; bcl-xL; apoptosis; nasopharyngeal carcinoma cells;
D O I
10.1111/j.1745-7254.2005.00529.x
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Aim: To evaluate a new plasmid mediated RNA interference (RNAi) system and investigate whether knock-down of bcl-xL by short hairpin RNA (shRNA) can induce apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2Z in vitro. Methods: The plasmid containing mU6 promoter was subcloned to yield the pmU6 plasmid, recombinant plasmid expressing shRNA targeting bcl-xL gene was designed and constructed, and were co-transfected cells with green fluorescence protein expressing plasmid. Flow cytometry was used to evaluate transfection efficiency, and RT-PCR and Western blot were applied to analyze bcl-xL mRNA and protein levels, respectively. Results: The shRNA expressed by the recombinant plasmid efficiently suppressed bcl-xL gene expression and induced apoptosis of NPC cells in vitro. Conclusion: The recombinant plasmid can sufficiently mediate RNAi in CNE-2Z cells, and knock-down of the bcl-xL expression by shRNA significantly induced apoptosis in CNE-2Z cells. The results suggest this new system, mediated RNAi can be used as a tool for the study of gene function and gene therapy.
引用
收藏
页码:228 / 234
页数:7
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