乙型肝炎病毒X蛋白通过miR-122调节肝癌细胞增殖及细胞周期的研究  

Hepatitis B Virus X Protein Regulates Proliferation and Cell Cycle of Hepatocellular Carcinoma Cells Through miR-122

作　　者：杨杰[1] 王永辉[1] 叶璟[1] 吴银霞[1] 虞作春[1] YANG Jie;WANG Yonghui;YE Jing;WU Yinxia;YU Zuochun(Lishui Central Hospital,Lishui 323000,China)

机构地区：[1]丽水市中心医院,丽水323000

出　　处：《病毒学报》2020年第1期70-76,共7页Chinese Journal of Virology

摘　　要：乙型肝炎病毒中的功能蛋白乙肝病毒X蛋白(Hepatitis B virus X protein,HBx)在促进肝细胞恶性改变中起到重要作用,但目前HBx调控肝癌细胞生长的具体机制仍未完全阐明。miR-122是具有抑癌特性的一类miR,在乙肝相关肝癌中表达减少。为了研究HBx通过微小RNA(microRNA,miR)-122调节肝癌细胞增殖及细胞周期的作用,本研究培养肝癌HepG2细胞株并进行分组,NC组转染NC慢病毒载体、HBx组转染HBx慢病毒载体、HBx+NC模拟物组转染HBx慢病毒载体及NC模拟物、HBx+miR-122模拟物组转染HBx慢病毒载体及miR-122模拟物、NC模拟物组转染NC模拟物、miR-122模拟物组:转染miR-122模拟物。通过MTS法检测细胞增殖活力,流式细胞术检测细胞周期,PCR检测miR-122表达量,western blot检测细胞周期蛋白G1(CyclinG1)、X连锁凋亡抑制蛋白(XIAP)、β-连环蛋白(β-catenin)的表达量。结果显示HBx组细胞的OD值、细胞周期G2/M期比例及细胞中CyclinG1、XIAP、β-catenin的表达量均明显高于NC组(P<0.05),细胞周期G0/G1期、S期比例及细胞中miR-122表达量均明显低于NC组(P<0.05);HBx+miR-122模拟物组细胞的OD值、细胞周期G2/M期比例及细胞中CyclinG1、XIAP、β-catenin的表达量均明显低于HBx+NC模拟物组(P<0.05),细胞周期G0/G1期、S期比例及细胞中miR-122表达量均明显高于HBx+NC模拟物组(P<0.05);miR-122模拟物组CyclinG1、XIAP、β-catenin荧光素酶报告基因的荧光活性明显低于NC模拟物组(P<0.05)。本研究结果充分说明HBx能够增强肝癌细胞的增殖活力及明显加速细胞周期,且该作用部分由miR-122的下调所介导。本研究首次阐明了HBx调节肝癌细胞生长的分子机制,也初步探明了具有抑癌活性的miR-122在肝癌细胞中可能靶向CyclinG1、XIAP、β-catenin等基因。Hepatitis B virus X protein(HBx),a functional protein of hepatitis B virus,plays an important role in promoting the malignant changes of liver cells. However,the mechanism of HBx in regulating the growth of liver cancer cells has not been fully elucidated. microRNA(mi-R)-122 is a kind of miR with anti-tumor properties,and its expression is decreased in hepatitis B-related liver cancer. To study the role of HBx in regulating the proliferation and cell cycle of hepatocellular carcinoma cells(HCCs)through miR-122,HepG2 cell lines were cultured and grouped. The NC group was transfected with the negative control(NC)lentivirus vector. The HBx group was transfected with the HBxlentivirus vector. The HBx+NC mimic group was transfected with the HBxlentivirus vector and NC mimic. The HBx+miR-122 mimic group was transfected with the HBxlentivirus vector and miR-122 mimic. The NC mimic group was transfected with the NC mimic.The miR-122 mimic group was transfected with the miR-122 mimic. Then cell proliferation was detected by MTS,cell cycle was detected by flow cytometry,expression of miR-122 was detected by PCR,cyclin G1,Xlinked inhibitor of apoptosis protein(XIAP)and beta-catenin were detected by Western Blot. Results showed that the optical density(OD),ratio of the G2/M phase of the cell cycle and expression of cyclinG1,XIAP,β-catenin in cells of the HBx group were significantly higher than those of the NC group. The ratio of the G0/G1 phase to S phase and expression of miR-122 in cells of the HBx group were significantly lower than those of the NC group. The OD,ratio of G2/M phase of the cell cycle and expression of cyclinG1,XIAP and β-catenin in cells of the HBx+mR-122 mimic group were significantly lower than those of the HBx+NC mimic group. The ratio of the G0/G1 phase to S phase and expression of miR-122 in cells of the HBx+mR-122 mimic group were significantly higher than those of the HBx+NC mimic group. The fluorescence activity of cyclin G1,XIAP andβ-catenin luciferase reporter genes in the miR-122 mimic group

关 键 词：肝癌 乙型肝炎病毒X蛋白(HBx) 微小RNA(miR)-122 增殖 细胞周期 

分 类 号：R373.2[医药卫生—病原生物学]