摘要
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 Xprotein,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。
Objective To investigate whether NF-κB 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-κB inhibitor pyrollidine dithiocarbamate（PDTC）was used to block the NF-κB signaling pathway.The activation and inactivation of NF-κB 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 using 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-κB 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（P0.05）.When the NF-κB 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（P0.05）.Conclusion NF-κB 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.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期54-58,63,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省科技计划资助项目(No.2009CDB117)
关键词
乙型肝炎病毒X蛋白
核因子-ΚB
肺耐药相关蛋白
肝癌
Hepatitis B virus X protein
nuclear factor-kappa B
lung resistance-related protein
hepatic carcinoma
