乙肝病毒x蛋白对肝癌细胞中PUMA表达的影响  

The effects of hepatitis B virus x protein on the expression of PUMA in hepatocellular carcinoma cells

作　　者：彭松林[1] 戴朝六[1] 姚殿波[1] Peng Songlin;Dai Chaoliu;Yao Dianbo(Department of Hepatobiliary Surgery,the Affiliated Shengjing Hospital of China Medical University,Liaoning Shenyang 110004,China.)

机构地区：[1]中国医科大学附属盛京医院肝胆脾外科,辽宁沈阳110004

出　　处：《现代肿瘤医学》2018年第11期1657-1662,共6页Journal of Modern Oncology

基　　金：沈阳市科学技术计划项目(编号:F13-221-9-24)

摘　　要：目的:探讨乙肝病毒x蛋白(hepatitis B virus x protein,HBx)对肝癌细胞中PUMA表达的影响及其机制。方法:利用质粒转染技术对p53基因不同状态的肝癌细胞转染野生型HBx基因,用RT-PCR检测转染后肝癌细胞中HBx mRNA表达和荧光定量PCR检测NFκB mRNA表达,Western blot检测HBx、p53、NFκB、PUMA蛋白表达。结果:转染后随着HBx的表达,在p53野生型肝癌细胞BEL-7402和p53突变型肝癌细胞Huh-7中NFκB mRNA和蛋白上调而p53蛋白表达下降,BEL-7402细胞PUMA蛋白表达下降而Huh-7细胞中PUMA蛋白表达略有下降。加入NFκB抑制剂PDTC后并随着PDTC浓度逐渐增加,BEL-7402细胞NFκB蛋白表达逐渐下降而p53蛋白和PUMA蛋白表达逐渐上升;但在Huh-7细胞中随着NFκB蛋白表达的抑制,p53蛋白逐渐上升而PUMA蛋白上升不明显。结论:结果提示转染HBx基因可以抑制肝癌细胞中PUMA蛋白表达,在p53野生型肝癌细胞这种抑制可能与HBx通过NFκB的激活有关。Objective：To investigate the effects of HBx protein on the expression of PUMA in hepatocellular carcinoma（ HCC） cells and its mechanism.Methods：Wild HBx plasmid was transfected into HCC cell line with different types of p53 gene.HBx mRNA expression was detected by RT-PCR.Fluorogenic quantitative PCR and Western blot were respectively administrated to the detection of nuclear factor kappa B（ NFκB） mRNA and proteins of HBx,p53,NFκB and PUMA.Results：The mRNA and protein of NFκB were upregulated and the p53 protein was down-regulated in BEL-7402 with p53-wild and in Huh-7 with p53-mutation following the transfection of HBx plasmid,and the expression of PUMA protein was down-regulated.In addition,the p53 protein and PUMA protein increased all following the inactivitation of NFκB by PDTC in BEL-7402.But the HBx-mediated PUMA inhibition was not removed following the inactivitation of NFκB in Huh-7.Furthermore,it was observed that HBx induced the increase of mRNA and protein of NFκB and reduced p53 protein in BEL-7402 and Huh-7 simultaneously.But p53 protein was increased simultaneously following the inactivitation of NFκB by PDTC.Conclusion：These data show that HBx induces the down-regulation of PUMA in HCC cells,which might be related to the inhibition of p53 via NFκB signaling in p53-wild HCC cells.

关 键 词：肝细胞癌 乙肝病毒X蛋白 p53上调凋亡调控因子 核因子ΚB P53 

分 类 号：R735.7[医药卫生—肿瘤]