论文部分内容阅读
目的检测外源基因HBx转入HepG2细胞后对PPARγ表达定量及定位的影响。方法以HBx重组腺病毒感染HepG2细胞,分别用RT-PCR和总蛋白Western blot检测PPARγ表达量的改变,免疫细胞化学检测其定位改变,Western blot分别检测细胞质和细胞核中PPARγ蛋白量的表达改变,MTT法检测HBx对曲格列酮抑制HepG2增殖效率的影响。结果外源基因HBx转入HepG2细胞后,RT-PCR、总蛋白Western blot显示PPARγ表达差异无统计学意义(P>0.05),免疫细胞化学及细胞质和细胞核中PPARγ蛋白检测提示PPARγ在细胞核内表达降低而在细胞质中出现积聚,曲格列酮对HepG2细胞增殖的抑制效率降低(P<0.05)。结论外源基因HBx对HepG2细胞中PPARγ的表达量无明显影响,但影响其向核内转移。HBx降低曲格列酮对HepG2细胞增殖的抑制效率。
Objective To detect the effect of exogenous HBx gene transfer into HepG2 cells on the quantitative and localization of PPARγ expression. Methods HepG2 cells were infected with HBx recombinant adenovirus. The expression of PPARγ was detected by RT-PCR and Western blot respectively. The localization of PPARγ was detected by immunocytochemistry. The expression of PPARγ in cytoplasm and nucleus was detected by Western blot. Effect of HBx on the Inhibition of HepG2 Proliferation by. Results After transfection of HBx into HepG2 cells, there was no significant difference in PPARγ expression between RT-PCR and Western blot (P> 0.05). Immunocytochemistry and PPARγ protein detection in cytoplasm and nucleus suggested that PPARγ was expressed in nucleus Decreased and accumulated in the cytoplasm. The inhibitory effect of troglitazone on HepG2 cell proliferation decreased (P <0.05). Conclusion The exogenous gene HBx has no obvious effect on the expression of PPARγ in HepG2 cells, but it affects the metastasis to the nucleus. HBx reduces the inhibitory effect of troglitazone on the proliferation of HepG2 cells.