HBx通过Wnt/β-catenin通路促进HepG2细胞增殖  

Hepatitis B Virus x Protein (HBx) Enhanced the Proliferation of HepG2 Cells Through Wnt/β-catenin Pathway

作　　者：郑碧云[1] 方雪芬 陈治新[1] 高雯宇 李丹[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化科,福州350001

出　　处：《福建医科大学学报》2016年第6期354-358,364,共6页Journal of Fujian Medical University

基　　金：国家自然科学基金青年项目(81300321);福建省财政专项经费(闽财指[2015]1297号);福建省自然科学基金青年项目(2016J05189);福建省卫计委中青年骨干重点项目(2014-ZQN-ZD-9);福建省临床医学重点专科资助项目(闽卫科教[2012]149号)

摘　　要：目的探讨Wnt/β-catenin通路在稳定表达乙型肝炎病毒x蛋白(HBx)的HepG2肝癌细胞增殖中的作用机制。方法验证前期构建的HepG2/HBx细胞株能稳定表达HBx蛋白。CCK-8法检测HepG2/HBx,HepG2/mock及HepG2 3组细胞增殖情况,RT-PCR及Western-blot法分别检测上述3种细胞中β-catenin mRNA和蛋白表达水平;最后检测β-catenin选择性抑制剂XAV939(20μmol/L)对各组细胞增殖水平的影响。结果前期构建的HepG2/HBx细胞株能稳定表达HBx蛋白。细胞增殖实验表明,HepG2/HBx组细胞增殖能力强于对照组(P<0.05)。RT-PCR及Western-blot检测结果显示,HepG2/HBx组细胞中β-catenin的mRNA及蛋白表达水平均高于对照组(P<0.05),而HepG2/mock及HepG2组细胞之间则无明显差别。XAV939能够抑制各组细胞的增殖能力,HepG2/HBx组细胞在加入20μmol/L XAV939作用后的增殖速度较加入相同浓度DMSO的HepG2/HBx组细胞下降(P<0.05)。XAV939对HepG2/HBx组细胞增殖的抑制强于对照组。结论 HBx蛋白可以上调肝细胞β-catenin表达,激活Wnt/β-catenin通路,促进肝癌细胞增殖。选择性β-catenin抑制剂可部分逆转该作用。Objective To observe the effect of Wnt/β-catenin pathway on the proliferation of HepG2 cells expressing hepatitis B virus x protein（HBx）stably. Methods We verified that HepG2/HBx cell lines constructed previously in our lab could stably express HBx protein. Then,cell-counting Kit-8（CCK-8）assay was used to detect the proliferation of HepG2/HBx,HepG2/mock and HepG2 cells. RT-PCR and Western-bolt were used to examine the level ofβ-catenin mRNA and protein expression in cells of the three cell groups. Finally,the level of cell proliferation was measured respectively after treated with 20μmol/Lβ-catenin selective inhibitor XAV939 for 24h. Results HepG2/HBx cell lines constructed previously in our lab could stably expressing HBx protein. CCK-8assay displayed that the proliferation rate of HepG2/HBx was higher than that of HepG2/mock and HepG2 cells.The level ofβ-catenin mRNA and protein expression was greater in HepG2/HBx cells compared with that of HepG2/mock and HepG2cells（P〈0.05）. β-catenin selective inhibitor XAV939 suppressed cell growth of the three groups. HepG2/HBx cells with XAV939 showed lower proliferation rate than that with DMSO. The proliferation inhibition rate of HepG2/HBx cells was significantly higher than that of other two groups at three time points（P〈0.05）. Conclusion The results indicate that HBx up-regulatesβ-catenin expression to activate wnt/β-catenin pathway and then promotes HepG2 cells proliferation.

关 键 词：病毒蛋白质类 乙型肝炎病毒X蛋白 细胞增殖 反式激活因子类 原癌基因 

分 类 号：R512.62[医药卫生—内科学]