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目的 研究乙型肝炎病毒x基因 (HBx)对肝癌细胞端粒酶活性及其催化亚单位端粒酶反转录酶 (TERT)表达的影响 ,探讨肝癌细胞端粒酶激活的分子机制。方法 分别应用端粒序列重复扩增法 (TRAP)和定量逆转录 聚合酶链反应 (RT PCR)检测QGY770 1肝癌细胞株 (n=5 )和转HBx基因肝癌细胞株QGY/HBx(n =5 )的端粒酶活性及TERTmRNA表达。结果 HBx阳性的QGY/HBx肝癌细胞TERT基因表达水平 1.5 4± 0 .14较HBx阴性的QGY770 1肝癌细胞0 .76± 0 .0 8明显增高 (P <0 .0 1) ;与之对应 ,QGY/HBx细胞的端粒酶活性也显著高于QGY770 1细胞 [( 4.2 8± 2 .81)× 10 5比 ( 2 .43± 1.46)× 10 5,P <0 .0 5 ]。结论 HBx基因可增强肝癌细胞端粒酶活性 ,上调TERT基因表达 ;HBx TERT途径可能是肝癌细胞端粒酶端粒激活的一个重要机制。
Objective To investigate the effect of hepatitis B virus x gene (HBx) on the telomerase activity and the expression of telomerase reverse transcriptase (TERT) in hepatocellular carcinoma cells, and to explore the molecular mechanism of telomerase activation in hepatoma cells. Methods QGY770 1 liver cancer cell line (n=5) and HBx gene hepatocellular carcinoma cell line QGY/HBx (n=5) were detected by TRAP and quantitative RT-PCR, respectively. ) Telomerase activity and TERT mRNA expression. Results The expression of TERT gene in HBx-positive QGY/HBx hepatoma cells was significantly higher than that of HBx-negative QGY7701 hepatocytes (1.54±0.14) (P < 0.01). The telomerase activity of QGY/HBx cells was also significantly higher than that of QGY770 1 cells [(4.28±2.81)×10 5 (2.43± 1.46) × 10 5, P <0.05]. Conclusion HBx gene can enhance the telomerase activity of hepatoma cells and up-regulate the expression of TERT gene. The HBx TERT pathway may be an important mechanism of telomerase activation of hepatoma cells.