Objective To investigate whether NF-kB signaling pathway is involved in the up-regulation of multi-drug resistant associated protein lung resistance-related protein(LRP)by hepatitis B virus X protein(HBx). Methods A cell line L02/HBx stably transfected with HBx was established. The NF-kB inhibitor pyrollidine dithiocarbamate(PDTC) was used to block the NF-kB signaling pathway. The activation and inactivation of NF-kB signaling pathway were examined by laser scanning confocal microscope(LSCM)before and after PDTC treatment. The levels of mRNA and protein expression of LRP were detected by u-sing real-time PCR and Western blot in non-transfected cell line L02 and stably transfected cell line L02/HBx when treated with or without PDTC. Results The NF-kB signaling pathway was activated after stable HBx gene transfection,and the expression levels of LRP mRNA and protein were increased by (2. 32 + 0. 31) and (4. 62 + 0. 72) times as compared with the control(P< 0. 05). When the NF-kB signaling pathway of L02/HBx cells was blocked by PDTC, the expression levels of LRP mRNA and protein were reduced by (1. 75 + 0. 19) and (2. 39 + 0. 54) times as compared with the control(P<0. 05). Conclusion NF-kB signaling pathway might be involved in the mechanism of HBx-mediated multi-drug resistance of hepatic carcinoma and the up-regulation of multi-drug resistance gene LRP.%目的 研究核因子-κB(nuclear factor-kappa B,NF-κB)信号通路是否是乙肝病毒X蛋白(hepatitis B virus X protein,HBx)上调多药耐药基因肺耐药相关蛋白(lung resistance-related protein,LRP)的途径之一.方法 建立稳定转染HBx基因的L02(L02/HBx)细胞系,用NF-κB信号通路阻断剂吡咯二硫代氨基甲酸酯(pyrollidine dithiocarbamate,PDTC)阻断NF-κB信号通路.采用荧光双标激光扫描共聚焦显微镜观察转染前后及PDTC加入前后NF-κB信号通路的激活失活情况,同时用Real-time PCR和Western blot检测转染前后及PDTC加入前后LRP的表达情况.结果 转染HBx基因后NF-κB信号通路被激活,LRP的mRNA和蛋白表达水平明显上调,分别为对照组的(2.32±0.31)倍和(4.62±0.72)倍(均P＜0.05);PDTC加入后NF-κB信号通路被阻断,LRP的mRNA和蛋白表达水平下调,分别为对照组的(1.75±0.19)倍和(2.39±0.54)倍(均P＜0.05).结论 NF-κB信号通路可能是HBx介导肝癌多药耐药,上调多药耐药基因LRP的途径之一.
展开▼
