论文部分内容阅读
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 investigate the effect of antisense human telomerase RNA (hTR) on the apoptosis of Bel-7402 hepatocellular carcinoma cell line and its molecular mechanism. Methods The human hepatoma cells transfected with human telomerase positive and antisense RNA were successfully transfected into human hepatocellular carcinoma cell line Bel-7402-hTR-EcoRI and Bel-7402-hTR-BamHI ) And blank control group (Bel-7402). The expression of p53, Bcl-2 and Bax mRNA and protein were detected by PCR. The expression of p53, Bcl-2 and Bax protein were detected by real-time fluorescence quantitative PCR and Western blot respectively. Results Compared with the cells in the positive control group and the blank control group, some of the cells in the antisense transfected group showed characteristic changes in apoptosis, the mRNA and protein expressions of p53 and Bax were significantly increased (P <0.05), while those of Bcl-2 mRNA and protein expression levels were significantly lower (P <0.05). Conclusion Antisense telomerase RNA can induce apoptosis of hepatocellular carcinoma cells by up-regulating the expression of p53 and Bax and down-regulating the expression of Bcl-2.