HBV X基因转染对HepG2肝癌细胞凋亡的影响及其机制

Effect of the Hepatitis B Virus X Gene Transfection on Apoptosis of HepG2 and Its Mechanism

目的 :研究HBVX基因转染对HepG2肝癌细胞凋亡及凋亡相关因子表达的影响。 方法 :用脂质体转染法将HBx真核表达载体pcDNA3 /HBx瞬时转入HepG2细胞 ,以未转染的HepG2细胞及转染空载体 pcDNA3 的细胞为对照。RT PCR法检测HBx基因的表达 ;MTT法检测各组细胞的增殖活性 ;TUNEL法检测各组的凋亡情况。β actin为内参 ,半定量RT PCR法检测凋亡相关基因Bax、Bcl xL、c myc的表达量变化。结果 :pcDNA3 X转染HepG2细胞后 ,RT PCR扩增出HBVX片段 ,空质粒组及正常对照组均未扩增出相应片段。HepG2细胞转染HBx基因后 ,相对于对照组细胞增殖能力明显下降 ,凋亡增多 ,差异有显著性意义 (P <0. 0 1) ;转染HBx的细胞Bax、Bcl xL、c mycmRNA相对表达量较转染空质粒组和未转染质粒组明显增高 ,差异有显著性意义 (P <0 . 0 5 )。结论 :成功将HBx基因转染入HepG2细胞 ,并在细胞中表达 ,转染HBx基因可同时上调Bax、Bcl xL、c mycmRNA表达 ,促进HepG2细胞凋亡 ,抑制增殖。

Objective: To investigate HBxs effect on apoptosis of HepG2 and its effect on expression of apoptosis factors. Methods: The HBV X gene eukaryon expression vector pcDNA 3 X was transiently transfected into HepG2 cell by lipid media transfection. Untrasfected HepG2 and HepG2 transfected with pcDNA 3 were used as control. The expression of HBx in HepG2 was identified by RT PCR. MTT and TUNEL were employed to detect proliferation and apoptosis of three groups. Semi quantified RT PCR was used to evaluate the expression of Bax, Bcl xL, c myc in three groups, and β actin was used as report gene.Results: Proliferative capacity in HepG2/pcDNA 3 X was obviously decreased, accompanied with high rate of apoptosis. Compared with HepG2 and HepG2/pcDNA 3 cell,expression of Bax, Bcl xL, c myc in HepG2/pcDNA 3 X were up regulated.Conclusion:HBx can up regulate either Bax, c myc and Bcl xL. This effection results in cell apoptosis and impairs proliferative capacity of cell.