摘要
目的:研究含有与EGFR基因序列相同的19nt序列RNAi质粒针对HepG2细胞表皮生长因子受体的RNA干扰效果。方法:用pSIREN-hE转染细胞,其质粒骨架为RNAi-Ready pSIREN-Shuttle Vector,含有可以被U6启动子转录成shRNA的寡核苷酸链,大小为69 nt,含有2段19nt的寡核苷酸,用于编码EGFR基因的RNA干扰的siRNA的正义链及反义链,评价该RNAi质粒对人肝细胞癌细胞株(HepG2)EGFR基因的mRNA表达抑制效果及对细胞凋亡的作用。结果:HepG2细胞EGFR基因的mRNA被明显抑制并伴有凋亡增加。结论:含有与EGFR基因序列相同序列RNA质粒对EGFR基因的RNA干扰可能是治疗肝细胞癌的一种有效手段。
Objective: To evaluate the RNAi efficiency of siRNA plasmid on the EGFR gene in a HepG2 cell line. Methods: By using a plasmid vector containing duplex of oligonucleotides, in which 19nt specific sequence of EG-FR expressing cassette to induce RNAi, the expression of endogenous EGFR in HepG2 human hepatocarcinoma cells was suppressed. The HepG2 cells were transfected with pSIREN-hE consisting of a RNAi-ready pSIREN-shuttle vector and duplex of oligonucleotides, which were built up of 69nt base pair, into a 19nt specific sequence of EGFR expressing cassette that can be transcripted by promoter U6. Results: The mRNA of EGFR in HepG2 human hepatocarcinoma cells was decreased and cell proliferation was reduced due to the induction of apoptosis. Conclusion: siRNA-mediated inhibition of EGFR gene may be a useful approach in the treatment of human hepatocarcinoma.
出处
《山东大学学报(医学版)》
CAS
北大核心
2007年第3期313-316,321,共5页
Journal of Shandong University:Health Sciences