干扰RNA对HepG2肝癌细胞内源性c-myc表达的抑制作用  

Suppression of c-myc expression by interference RNA in HepG2 hepatocellular carcinoma cells

作　　者：许杨[1] 王益华[1] 高纪东[1] 叶珏[1] 朱红霞[1] 徐宁志[1] 王兴宇[1] 孙宗棠[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤医院肿瘤研究所分子肿瘤学国家重点实验室,北京100021

出　　处：《中华肿瘤杂志》2004年第8期458-460,共3页Chinese Journal of Oncology

基　　金：国家重点基础研究发展规划课题资助项目(G19980 5 12 0 9)

摘　　要：目的　研究干扰RNA(RNAi)对HepG2细胞的c myc基因表达的干扰效应。方法　建立装载靶向c myc基因小干扰性RNA(siRNA)的表达载体。用脂质体法将此表达载体转染HepG2细胞株 ,以转染空载细胞作为对照。采用定量PCR和Westernblot检测c myc基因的表达。用流式细胞仪及荧光显微镜检测AnexinV标记的凋亡细胞。结果　转染siRNA的表达载体可以抑制内源c myc基因在转录和转译上的表达 ,抑制率达 6 7%。c myc基因的表达抑制与HepG2细胞的凋亡相关联。结论　转染siRNA的表达载体可明显抑制肝癌细胞株HepG2c myc基因的表达 。Objective To study the inhibitory effect of RNA interference (RNAi) on c-myc expression in hepatocellular carcinoma cell line, HepG2. Methods Expression vector of c-myc gene-targeting small interference RNA (siRNA) was constructed (psilencer-c-myc) and transfected into HepG2 cells by lipofectamine, and the unloaded vector was used as control (mock). The expression of c-myc mRNA and protein was identified by quantitive PCR and Western blot. Apoptosis of the transfected cells was examined by flow cytometry and immunofluorescent microscopy. Results After HepG2 cells were transfected with psilencer-c-myc, the expression of c-myc mRNA and protein was suppressed with an inhibition rate of 67% compared with the mock-transfected cells. Apoptosis was identified in the transfected HepG2 cells. Conclusion The expression of c-myc at transcriptional and translational levels in HepG2 cells transfected with siRNA is markedly inhibited, which may be associated with the induction of apoptosis.

关 键 词：干扰RNA HEPG2 肝癌细胞 内源性c-myc表达 抑制作用 肝细胞 

分 类 号：R735.7[医药卫生—肿瘤]