乙型肝炎病毒通过miR⁃154⁃5p/STAT3轴促进肝癌细胞的增殖和迁移  

Hepatitis B Virus Promotes the Proliferation and Migration of Liver Cancer Cells Through the MiR⁃154⁃5p/STAT3 Axis

作　　者：高瑞娜[1] 赵晓燕[1] 杨文静[1] 李鹏丽 GAO Ruina;ZHAO Xiaoyan;YANG wenjing;LI Pengli(Shuozhou Vocational Technical College,Shuozhou 036002,China;Taiyuan Central Hospital,Taiyuan 030002,China)

机构地区：[1]朔州职业技术学院,朔州036002 [2]北京大学第一医院太原医院,太原030002

出　　处：《病毒学报》2024年第4期750-756,共7页Chinese Journal of Virology

摘　　要：探讨乙型肝炎病毒(Hepatitis B virus,HBV)通过miR⁃154⁃5p/STAT3轴促进肝癌细胞增殖和迁移的影响。用含有1.3倍超长HBV基因组的腺病毒感染HepG2细胞,以正常HepG2作为对照(NC),记为HBV组和NC组。采用实时荧光定量PCR(qRT⁃PCR)检测HBV组、NC组、稳定表达HBV的人肝癌细胞HepG2.2.15及人肝癌细胞HepG2中miR⁃154⁃5p表达水平。按照脂质体法将miR⁃NC模拟物、过表达miR⁃154⁃5p、过表达miR⁃154⁃5p+空载体pcDNA、过表达miR⁃154⁃5p+过表达STAT3转染至HepG2.2.15细胞中,记为miR⁃NC组、miR⁃154⁃5p组、miR⁃154⁃5p+pcDNA组、miR⁃154⁃5p+STAT3组。细胞计数试剂盒(CCK8)、克隆形成实验检测细胞增殖;伤口愈合实验检测细胞迁移;蛋白免疫印迹(Western blot)法检测信号转导与转录因子3(STAT3)、增殖细胞核抗原(PCNA)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)蛋白表达。荧光素酶实验检测miR⁃154⁃5p和STAT3的关系。与NC组相比,HBV组内miR⁃154⁃5p表达水平降低;与HepG2细胞相比,HepG2.2.15细胞中miR⁃154⁃5p表达水平明显降低。上调miR⁃154⁃5p明显降低细胞活性、克隆形成数、迁移率,并抑制了PCNA、MMP2和MMP9蛋白表达。miR⁃154⁃5p靶向负调控STAT3的表达,且过表达STAT3可以逆转上调miR⁃154⁃5p对HepG2.2.15细胞增殖、迁移的抑制作用。乙型肝炎病毒通过降低miR⁃154⁃5p表达上调STAT3,进而促进肝癌细胞的增殖和迁移。To investigate the effect of hepatitis B virus(HBV)on the proliferation and migration of liver cancer cells through the miR⁃154⁃5p/STAT3 axis.HepG2 cells were infected with adenovirus containing 1.3⁃fold overlength HBV genome,and normal HepG2 was used as control(NC),which was denoted as HBV group and NC group.Quantitative real⁃time PCR(qRT⁃PCR)was used to detect the expression level of miR⁃154⁃5p in HBV group,NC group,human hepatocellular carcinoma cell HepG2.2.15 with stable HBV expression and human hepatocellular carcinoma cell HepG2.Transfected miR⁃NC mimic,overexpression miR⁃154⁃5p,overexpression miR⁃154⁃5p+empty vector pcDNA,overexpression miR⁃154⁃5p+overexpression STAT3 into HepG2.2.15 cells according to the liposome method,denoted as miR⁃NC group,miR⁃154⁃5p group,miR⁃154⁃5p+pcDNA group,miR⁃154⁃5p+STAT3 group.Cell counting kit(CCK8),clone formation test to detect cell proliferation;wound healing test to detect cell migration;Western blot method to detect signal transduction and transcription factor 3(STAT3),proliferating cell nuclear antigen(PCNA),matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)protein expression.The luciferase experiment detects the relationship between miR⁃154⁃5p and STAT3.Compared with the NC group,the expression level of miR⁃154⁃5p in the HBV group was reduced;compared with the HepG2 cells,the expression level of miR⁃154⁃5p in the HepG2.2.15 cell was significantly reduced.Up⁃regulation of miR⁃154⁃5p significantly reduced cell viability,number of clones,migration rate,and inhibited PCNA,MMP2,and MMP9 protein expression.miR⁃154⁃5p targets and negatively regulates the expression of STAT3.Overexpression of STAT3 can reverse the inhibiting effect of miR⁃154⁃5p up⁃regulation on the proliferation and migration of HepG2.2.15 cells.HBV inhibits miR⁃154⁃5p expression to upregulate STAT3,thereby promoting the proliferation and migration of liver cancer cells.

关 键 词：乙型肝炎病毒 miR⁃154⁃5p STAT3 肝癌细胞 增殖 迁移 

分 类 号：R735.7[医药卫生—肿瘤]