乙型肝炎病毒P22e蛋白经核因子-κB抑制HepG2细胞凋亡

目的 探讨核因子-κB(NF-κB)在乙型肝炎病毒P22e蛋白抑制HepG2细胞凋亡中的作用.方法 用含HBV P22e基因的重组pEGFP-C2HBVP22e质粒的肝癌细胞HepG2,以放线菌素-D(Act-D)、肿瘤坏死因子(TNF)α诱导该细胞凋亡,采用激光共聚焦显微镜、核蛋白电泳迁移率等技术,观察在HBV P22e抑制TNFα诱导HepG2EGFP-C2HBVP22e细胞的凋亡过程中NF-κB的核转移、活化等情况.用NF-κB抑制剂ALLN抑制其信号通路,检测以Act-D、TNFα诱导的HepG2、HepG2EGFP-C2HBVP22e细胞凋亡率的变化.对实验结果的数据分析用秩和检验和t检验. 结果激光共聚焦显微镜及电泳迁移率实验观察到HepG2EGFP-C2HBVP22e细胞在发生凋亡前后,有明显的NF-κB向核内迁移活化现象.NF-κB抑制剂ALLN可使以Act-D、TNF α诱导HepG2EGFP-C2HBVP22e细胞的凋亡率明显升高(6.19%±1.58%与39.99%±7.620/0,t=7.515,P<0.01).结论 在HBV P22e蛋白抑制肝癌细胞凋亡过程中,NF-κB信号途径起着重要作用.

Hepatitis B virus P22e inhibit hepatocyte apoptosis via nuclear factor kappa B

Objective To test whether nuclear factor kappa B plays an important role in the apoptosis-inhibitory effect of hepatitis B virus (HBV) P22e protein. Methods HepG2 cells were transfeeted with recom-bination plasmid pEGFP-HBVP22e. The Aet-D and TNF a were used to induce apoptosis. NF-κ B inhibitor ALLN were used to inhibit the signaling pathway. The activation of NF-κB was EMSA, and the nulear translo-cation of NF-κB was determined by immtmo-staining. Results Laser scanning eonfocai microscopy and EMSA indicated that HBV P22e protein enhanced the nuclear translocation ofNF-κB after apoptosis induction. ALLN treatment inhibited the nuclear translocation of NF-κB, and blocked the apoptosis-inhibiting effect of HBV P22e protein. Conclusion This study indicates that HBV P22e protein inhibits apoptosis ofhepatocyte via the NF-κB signaling pathway.