乙型肝炎病毒X基因在体内外对肝癌细胞增殖活性的影响

HBx gene facilitates the proliferation activity of hepatoma cells in vitro and in vivo

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 effects of HBx gene on proliferation activity of hepatoma cells in vitro and in vivo. Methods The plasmid pHA-HBx carrying HBx gene was transfected into HepG_2 cells,and the positive clones were screened and identified with G418 and RT-PCR,respectively.The growth curve and population doubling time were calculated,and the cell cycle was analyzed by flow cytometry (FCM).The proliferation activity of transformed cells was measured with 3H-TdR incorporation rate and nude mice model in vitro and in vivo. Results The result of RT-PCR indicated that HBx gene was integrated into the genome DNA of HepG_2 cells and transcripted.The growth curve and population doubling time showed a high proliferation activity of transformed cells.The amount of cells at stage S and G_2/M were significantly higher,and cells at stage G_0/G_1 were lower than those in control group.The tumors developed from transfected cells grew much quicker than those developed from HepG_2 cells in nude mice model.Conclusion HBx gene can facilitate the proliferation of hepatoma cells both in vitro and in vivo.