HBV对HOXA10表达的影响及其机制研究  

Influence and mechanism of hepatitis B virus on the expression of HOXA10

作　　者：宋惠[1] 杨兰 徐莉敏[1] 沈振华[1] 刘倩倩 刘兴晖[1] SONG Hui;YANG Lan;XU Limin;SHEN Zhenhua;LIU Qianqian;LIU Xinghui(Department of Clinical Laboratory,Shanghai Gongli Hospital,Shanghai 200135,China)

机构地区：[1]上海市浦东新区公利医院检验科,上海200135

出　　处：《检验医学》2021年第11期1172-1176,共5页Laboratory Medicine

基　　金：上海市浦东新区卫生系统领先人才培养计划(PWRl2018-05);浦东新区卫生系统重点学科群建设资助项目(PWZxq2017-15)。

摘　　要：目的探讨乙型肝炎病毒(HBV)对同源框蛋白A10(HOXA10)表达的影响及其机制。方法选取慢性乙型肝炎(CHB)患者20例(CHB组)、体检健康者23名(正常对照组),分离其外周血单个核细胞(PBMC)。采用实时荧光定量聚合酶链反应(RT-qPCR)和免疫印迹法检测肝癌细胞系HepG2、HepG2.2.15及CHB组、正常对照组PBMC中HOXA10mRNA和蛋白的表达。将HBV单个基因的表达质粒(pCMV-S、pCMV-E、pCMV-C、pCMV-X、pCMV-P)分别与含有HOXA10基因启动子的荧光素酶报告基因pGL3-HOXA10共转染HepG2细胞,并测定荧光素酶活性。结果HepG2.2.15细胞中HOXA10mRNA和蛋白的相对表达量均显著高于HepG2细胞(P=0.001)。HBV患者PBMC中HOXA10 mRNA的相对表达量显著高于正常对照组(P=0.001)。转染pCMV-X的HepG2细胞的荧光素酶活性显著高于转染对照载体pCMV-tag2B的HepG2细胞(P=0.001),转染其他质粒的HepG2细胞之间荧光素酶活性差异均无统计学意义(P>0.05)。转染pCMV-X的HepG2细胞的HOXA10 mRNA和蛋白的相对表达量显著高于转染空载体pCMV-tag2B的HepG2细胞(P=0.001)。结论HBV可能通过其X基因上调HOXA10的表达。Objective To investigate the influence of hepatitis B virus(HBV)on the expression of homeobox protein A10(HOXA10)and its mechanism.Methods Totally,20 patients with chronic hepatitis B(CHB)(CHB group)and 23 healthy subjects(healthy control group)were enrolled,and peripheral blood mononuclear cells(PBMC)were isolated.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and western blotting were used to determine the expression of HOXA10 mRNA and protein in HepG2.2.15 cells,CHB group PBMC and healthy control group PBMC.The expression plasmids of HBV single gene(pCMV-S,pCMV-E,pCMV-C,pCMV-X and pCMV-P)were co-transfected with pGL3-HOXA10(a luciferase reporter gene which harbored HOXA10 gene promoter)into HepG2 cells,respectively,and the changes in luciferase activity were determined.Results The expression levels of HOXA10 mRNA and protein in HepG2.2.15 cells were higher than those in HepG2 cells(P=0.001).The expression of HOXA10 mRNA in the PBMC of HBV patients was higher than that of healthy control group(P=0.001).The luciferase activity of HepG2 cells transfected with pCMV-X was higher than that of HepG2 cells transfected with control vector pCMV-tag2 B(P=0.001).There was no statistical significance in luciferase activity between HepG2 cells transfected with other plasmids(P>0.05).The expression levels of HOXA10 mRNA and protein in HepG2 cells transfected with pCMV-X were higher than those in HepG2 cells transfected with control vector pCMV-tag2 B(P=0.001).Conclusions HBV might up-regulate the expression of HOXA10 through its X gene.

关 键 词：同源框蛋白A10 乙型肝炎病毒 X基因 

分 类 号：R446.1[医药卫生—诊断学]