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目的 探讨乙型肝炎病毒x(HBx)基因对肝癌细胞增殖活性的影响。方法 将携带HBx基因的表达质粒 pHA HBx转染HepG2 肝癌细胞 ,G4 18筛选阳性细胞克隆 ,RT PCR鉴定HBx基因的整合及表达 ;通过细胞计数 ,观察HBx基因对肝癌细胞生长曲线和倍增时间的影响 ;流式细胞术检测细胞周期的变化 ;3 H TdR掺入法检测细胞增殖活性 ;裸鼠接种观察HBx基因在体内对肝癌细胞增殖的影响。结果 HBx基因在体内外对HepG2 细胞的增殖活性均有明显影响 ,细胞生长曲线左移 ,倍增时间缩短 ;G0 /G1期细胞减少 ,S期和G2 /M期细胞增多 ;转染后的细胞3 H TdR掺入率较对照组增高 ;转HBx基因的裸鼠移植瘤的生长速度较对照组细胞明显加快。结论 HBx基因在体内外均可提高肝癌细胞的增殖活性 ,增加肝癌细胞的恶性表型 ,加速肿瘤生长。
Objective To investigate the effect of hepatitis B virus (HBx) gene on the proliferation of hepatoma cells. Methods HBx gene was transfected into HepG2 hepatocellular carcinoma cells with pHA HBx expression plasmid. Positive cells were screened by G418, and the integration and expression of HBx gene were identified by RT PCR. The effects of HBx gene on growth curve and doubling time of hepatocellular carcinoma cells were observed by cell counting The changes of cell cycle were detected by flow cytometry. The cell proliferation activity was detected by 3 H TdR incorporation method. The nude mouse was inoculated to observe the effect of HBx gene on the proliferation of hepatocellular carcinoma cells in vivo. Results The HBx gene had a significant effect on the proliferation of HepG2 cells both in vitro and in vivo. The cell growth curve shifted to the left and the doubling time was shortened. The number of cells in G0 / G1 phase decreased and the number of cells in S phase and G2 / M phase increased. H TdR incorporation rate higher than the control group; HBx gene transplanted tumor growth rate in nude mice significantly faster than the control group cells. Conclusion The HBx gene can enhance the proliferation of hepatoma cells in vitro and in vivo, increase the malignant phenotype of hepatocellular carcinoma cells and accelerate the growth of the tumor.