转HBx基因肝癌细胞增殖与抗凋亡特性研究

:【目的】研究转HBx基因肝癌细胞增殖与拮抗凋亡特性 ,探讨肝癌恶性表型的分子基础。【方法】以携HBx基因重组逆转录病毒感染QGY770 1肝癌细胞 ,G418筛选 ,PCR与RT PCR鉴定 ;流式细胞仪 (FCM)检测细胞增殖周期 ;用阿霉素诱导细胞凋亡 ,FCM定量检测。【结果】QGY770 1肝癌细胞经HBx基因转染后 ,G418筛选 4～ 6周获阳性克隆株QGY/HBx ,PCR与RT -PCR分别证实细胞有HBx基因整合与HBxmRNA表达 ;细胞周期分析结果表明 ,QGY/HBx细胞的细胞周期进程明显加快 ,并可明显抵制阿霉素的凋亡诱导作用。【结论】HBx基因可加速肝癌细胞周期的进程 ,并增强肝癌细胞的抗凋亡能力。

Studies on Characteristics of Proliferation and Apoptosis Resistance in Hepatocellular Carcinoma Cells with HBx Gene Transferred

Objective To study the effects of transferred HBx gene on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells for investigation of the molecular basis of malignancy behavior of HCC. Methods QGY7701 HCC cells were infected with recombinant retrovirus carrying HBx gene and then selectively cultured with G418 and identified by PCR and RT-PCR techniques. The cell cycle and apoptosis induced by adriamycin (ADM) were determined by flow cytometer (FCM) respectively. Results QGY7701 cells with HBx gene transferred were selectively cultured with G418 and the positive clones QGY/HBx obtained in 4 to 6 weeks. A HBx gene integration was detected by PCR in the genomic DNA of QGY/HBx cells and a steady expression of HBx mRNA by RT-PCR. The process of cell cycle of QGY/HBx cells were faster than QGY7701 cells. The QGY/HBx cells could resist the apoptosis induced by adriamycin. Conclusions HBx gene could improve the proliferation ability of HCC cells. HCC cells modified by HBx gene could resist the apoptosis induced by adriamycin.