反义人端粒酶RNA诱导肝癌细胞凋亡的分子生物学机制研究  

Study of the mechanism of apoptosis of human hepatocellular carcinoma cells induced by the antisense human telomerase RNA gene

作　　者：姜雅堃[1] 刘吉勇[1] 韩翠萍[1] 文廷玉[1] 刘瑾[1] 

机构地区：[1]山东大学附属省立医院消化内科,济南250021

出　　处：《山东大学学报（医学版）》2011年第12期25-29,共5页Journal of Shandong University:Health Sciences

基　　金：山东省医药卫生科研项目(2001CA1DBA3)

摘　　要：目的探讨反义人端粒酶RNA(human telomerase RNA,hTR)基因对肝癌细胞株Bel-7402凋亡的诱导作用及其分子生物学机制。方法利用前期实验成功转染人端粒酶正、反义RNA基因的肝癌细胞,分为正义转染组(Bel-7402-hTR-EcoRI)、反义转染组(Bel-7402-hTR-BamHI)和空白对照组(Bel-7402)。PCR鉴定转染结果,ho-echst 33258荧光染色观察3组细胞的变化,实时荧光定量PCR法和Western blot法分别检测凋亡相关基因p53、Bcl-2和Bax的mRNA和蛋白的表达水平。结果与正义转染组及空白对照组细胞相比,反义转染组部分细胞出现凋亡特征性改变,p53、Bax的mRNA和蛋白表达水平明显升高(P<0.05),Bcl-2的mRNA和蛋白表达水平明显降低(P<0.05)。结论反义端粒酶RNA可诱导肝癌细胞凋亡,其机制可能是通过上调p53和Bax的表达并下调Bcl-2的表达。Objective To study the mechanism of apoptosis of hepotocellular carcinoma cells induced by antisense human telomerase RNA（hTR）.Methods We previously transfected the hTR gene in its sense and antisense orientations into hepatocellular carcinoma cells.In the present work,three cell groups were used： the sense transfected group（BEL-7402-hTR-EcoRI）,the antisense transfected group（BEL-7402-hTR-BamHI） and the blank control group（BEL-7402）.PCR was done to verify the integration of the hTR gene.hoechst 33258 fluorescent labeling was carried out to detect cell apoptosis.Real-time fluorescent quantitative PCR and Western blot were performed to test expression of p53,Bax and Bcl-2 mRNA and protein,respectively.Results Compared with the blank control and the sense hTR transfected groups,the antisense hTR transfected groups showed typical apoptotic changes： expression of mRNA and protein of both p53 and Bax was significantly increased（P0.05）,and expression of mRNA and protein of Bcl-2 was significantly decreased（P0.05）.Conclusion Antisense human telomerase RNA can induce apoptosis of hepatocellular carcinoma cells.This is likely achieved by up-regulating expression of p53 and Bax and down-regulating expression of Bcl-2.

关 键 词：端粒酶 反义RNA 肝癌 细胞凋亡 基因 

分 类 号：R735.7[医药卫生—肿瘤]