丙型肝炎病毒NS4B蛋白调控Hep3B细胞非折叠蛋白质反应研究  

Regulation of Unfolded Protein Response in Hep3B Cells by HCV NS4B

作　　者：张艳妮[1] 张庆华[1] 陈瑶[1] 刘好桔[1] 刘志文[1] 孔令保[1] 

机构地区：[1]江西农业大学生物科学与工程学院,江西南昌330045

出　　处：《江西农业大学学报》2012年第1期165-169,共5页Acta Agriculturae Universitatis Jiangxiensis

基　　金：江西省科技支撑计划重点项目(2009BNA09400);江西省自然科学基金项目(2010GZY0059);江西农业大学校基金项目(2950)

摘　　要：研究表达的丙型肝炎病毒(HCV)NS4B对Hep3B细胞非折叠蛋白质反应的影响。NS4B重组真核表达质粒pcDNA3.1(-)NS4B通过脂质体转染Hep3B细胞,G418筛选和Western blot鉴定稳定转染细胞;RT-PCR检测稳定转染细胞内XBP1 mRNA剪接,Western blot鉴定ATF6蛋白切割,荧光素酶试验检测稳定转染细胞内GRP78和XBP1启动子活性。G418筛选和Western Blot鉴定证实获得稳定表达NS4B的Hep3B细胞;在该细胞内,XBP1 mRNA剪接、ATF6切割、XBP1和Grp78启动子激活均被检测到。NS4B在Hep3B的稳定表达诱导了非折叠蛋白质反应。To study the effect of hepatitis C virus NS4B on unfolded protein response in Hep3B cells. The recombinant vector pcDNA3. 1 （ - ） NS4B with NS4B gene was transfected to Hep3B cells using Lipo- fectamine 2000. Stably transfected cells were selected by G418 and then confirmed by Western blot analysis; XBP1 mRNA splicing, ATF6 cleavage, GRP78 and XBP1 promoter activation were analyzed in stably trans- fected Hep3B cells using RT - PCR, Western blot or luciferase assays. G418 seclection and Western blot demonstrated that Hep3B cells stably expressing NS4B were obtained successfully; XBP1 mRNA splicing, ATF6 cleavage,GRP78 and XBP1 promoter activation were observed in Hep3B cells stably expressing NS4B. Stable expression of NS4B in Hep3B cells induces unfolded protein response.

关 键 词：丙型肝炎病毒 NS4B HEP3B 非折叠蛋白质反应 

分 类 号：R373[医药卫生—病原生物学]