半乳糖凝集素-3结合蛋白作为正向调节因子参与IFN-γ介导的抗HBV作用  

LGALS3BP Acts as a Positive Regulator and is Involved in IFN-γMediated Anti-HBV Process

作　　者：孟诗敏 陈忠伟 李涵泺 魏艳红 胡康洪 MENG Shi-min;CHEN Zhong-wei;LI Han-luo;WEI Yan-hong;HU Kang-hong(School of Bioengineering and Food Science,Hubei Provincial Key Laboratory ofIndustrial Microbiology,Sino-German Biomedical Center,National'111'Center for Cellular Regulation and Molecular Pharmaceutics,Hubei University of Technology,Wuhan,Hubei,430068,China)

机构地区：[1]湖北工业大学生物工程与食品学院中德生物医学中心工业微生物湖北省重点实验室国家外专局/教育部细胞调控与分子药物"111"引智基地,湖北武汉430068

出　　处：《现代生物医学进展》2021年第23期4441-4449,4535,共10页Progress in Modern Biomedicine

基　　金：湖北工业大学高层次人才启动基金项目(337203)。

摘　　要：目的:研究半乳糖凝集素-3结合蛋白(LGALS3BP)在IFN-γ介导的抗HBV中的作用。方法:采用qRT-PCR及Western Blot分别检测HepG2细胞瞬转过表达质粒及干扰质粒后LGALS3BP转录水平和翻译水平的变化及检测IFN-γ对HepG2细胞内源性LGALS3BP转录水平和翻译水平的影响;采用CCK-8评价LGALS3BP表达量的变化对HepG2细胞增殖的影响及确定IFN-γ处理HepG2和HepG2.215的最适浓度;采用ELISA检测当LGALSEBP表达量发生变化时,IFN-γ刺激后对细胞外分泌的HBs Ag和HBeAg的影响;采用q-PCR和RT-PCR分别检测细胞内HBV核衣壳中DNA和细胞内HBV RNA水平。结果:40ng/m L的IFN-γ处理HepG2细胞48小时,LGALS3BP m RNA表达量与对照组相比提高2.87倍,但LGALS3BP表达量的变化对细胞增殖无显著作用;当过表达LGALS3BP时,在IFN-γ介导的抗HBV过程中,与对照组相比,HepG2.215细胞分泌的HBs Ag、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为46%、67%、59%、49%;与对照组相比,转染pCH9-3091的HepG2细胞分泌的HBs Ag、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为67%、53%、60%、29%;而当干扰LGALS3BP时,在IFN-γ介导的抗HBV过程中,与对照组相比,HepG2.215细胞分泌的HBs Ag、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为46%、67%、59%、49%;与对照组相比,转染pCH9-3091的HepG2细胞分泌的HBs Ag、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为67%、77%、67%、45%。结论:LGALS3BP作为正向调节因子参与IFN-γ介导的抗HBV过程。Objective:This study was to investigate the role of LGALS3 BP in the IFN-γmediated anti-HBV process in human hepatocytes in vitro.Methods:HepG2 and HepG2.215 cell lines were transiently transfected with LGALS3 BP overexpression plasmid and its interfering plasmid,and levels of LGALS3 BP transcription and translation were detected by means of qRT-PCR and Western Blot.CCK-8 was used to determine the optimal concentration of IFN-γtreatment for HepG2 and HepG2.215 cells,and evaluate the cell proliferation.The effects of IFN-γtreatment on transcription and translation of endogenous LGALS3 BP in HepG2 cells were investigated using q RT-PCR and Western Blot.When changing LGALS3 BP expression,the effect of IFN-γstimulation on the exocrine HBs Ag and HBeAg were examined using ELISA,and levels of DNA in HBV nucleocapsid and intracellular RNA were evaluated using q-PCR and RT-PCR,respectively.Results:After treated with 40 ng/ml IFN-γfor 48 hours,the level of LGALS3 BP m RNA in HepG2 cells was 2.87 folds higher than that in the control group,whereas the change of LGALS3 BP expression had no significant effect on the cell proliferation.In case that LGALS3 BP was overexpressed in HepG2.215 cells during the IFN-γmediated anti-HBV process,compared with the control group,it was found the relative decrease rates of extracellular HBs Ag,HBeAg as well as of the relative decrease rates of intracellular HBV DNA-and RNA levels were 46%,67%,59%and 49%,respectively;For HepG2 cells that were transiently transfected with pCH9-3091,compared with the control group,the relative decrease rates of extracellular HBs Ag,HBeAg as well as the intracellular viral DNA and RNA were 67%,53%,60%and 29%,respectively.In contrast,in case that LGALS3 BP was down-regulated by si RNA interfering-plasmid SHLG,in the process of IFN-γmediated anti HBV for HepG2.215,compared with the control group,the relative increase rates of secreted HBs Ag,HBeAg as well as of intracellular viral DNA-and RNA levels were 46%,67%,59%and 49%,respectively;For HepG2 that tra

关 键 词：乙型肝炎病毒 干扰素Γ 半乳糖凝集素-3结合蛋白 增殖 

分 类 号：R-33[医药卫生] R373.21