丙型肝炎病毒抑制DNA损伤修复相关基因生长阻滞和DNA损伤诱生蛋白45α的表达  

Hepatitis C virus strain JFH1 down-regulates expression of growth arrest and DNA damage-inducible gene 45a in human hepatoma Huh7.5.1 cells

作　　者：成镀[1] 蒋永芳[1] 肖新强[1] 龚国忠[1] 

机构地区：[1]中南大学湘雅二医院感染科中南大学肝病研究所,长沙410011

出　　处：《中华肝脏病杂志》2012年第11期807-810,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30471531）;国家传染病“十一五”重大专项（2008ZX10002-013）

摘　　要：目的,探讨HCV感染对DNA损伤修复相关基因生长阻滞和DNA损伤诱生蛋白45仅（GADD45α）表达的影响。方法建立全基因HCVJFH1感染的Huh7．5．1细胞模型。用相对荧光定量PCR和Westernblot分别检测HCVJFHl感染和未感染的Huh7．5．1细胞中GADD45仅mRNA和蛋白质的表达水平。组间数据比较用单因素方差分析。结果HCVJFHl感染的Huh7．5．1细胞内有HCVRNA高水平复制及HCVNS5A蛋白质和核心蛋白的表达。与未感染Huh7．5．1细胞相比，HCVJFH1感染72h的细胞内GADD450α mRNA和蛋白质相对表达量均明显降低，分别为0．57±0．09比1．00±0．11和0．28±0．03比1．00±0．07，差异均有统计学意义（F值分别为75．407和560．04，P值均〈0．01）。结论HCV下调GADD45α的转录和蛋白质表达，影响DNA损伤修复，这可能是HCV感染致肝癌发生的机制之一。Objective To investigate the effect of hepatitis C virus （HCV） strain JFH1 on expression of the human gene, growth arrest and DNA damage-inducible gene 45α （GADD45α）, in infected hepatoma cells. Methods HCV JFH1 RNA-containing supematants were used to infect the human hepatoma cell line, Huh7.5.1; infection was confirmed by Western blot detection of the HCV-encoded non-structural 5A （NS5A） protein and core protein. Infection-induced changes in GADD45α mRNA and protein expressions were measured by real time PCR using SYBR Green and Western blotting, respectively. Significance of differences between the levels detected in JFHl-infected or uninfected Huh7.5.1 cells was analyzed by single factor analysis of variance testing. Results The HCV infection system was successfully established, as evidenced by expression of NS5A protein and core protein. The GADD45α mRNA and protein levels were significantly down- regulated in JFHl-infected Huh7.5.1 cells, by 0.57 ± 0.09 and 0.28 ±0.03, respectively, as compared to levels in uninfected Huh7.5.1 cells （F values were 75.407 and 560.04, respectively; P 〈 0.01）. Conclusion HCV inhibits the mRNA transcription and protein expression of host GADD45α, which may contribute to the pathogenesis of hepatocellular carcinoma caused by HCV infection.

关 键 词：肝炎病毒 DNA修复 生长阻滞和DNA损伤诱生蛋白45α 

分 类 号：R51[医药卫生—内科学]