HBx基因及其缺失突变体(HBx-d382)对L02细胞microRNA表达谱的影响  

Influence of HBx gene and its deletion mutation(HBx-d382) on microRNA expression profile in L02 cells

作　　者：胡志亮[1] 谭德明[1] 侯周华[1] 刘国珍[1] 谢萍[1] 欧阳奕[1] 刘菲[1] 刘洪波[1] 

机构地区：[1]中南大学湘雅医院感染科,湖南长沙410008

出　　处：《中国现代医学杂志》2010年第17期2561-2566,2569,共7页China Journal of Modern Medicine

基　　金：湖南省自然科学基金(No:10JJ5034);湖南省科技厅科技计划资助项目(No:2010SK3093)

摘　　要：目的研究转染HBx基因(hepatitis Bvirus Xgene)及其缺失突变体(HBx-d382)后L02细胞Mi-croRNA表达谱的变化。方法通过脂质体转染和G418筛选获得L02/HBx、L02/HBx-d382阳性克隆,逆转录聚合酶链式反应(RT-PCR)和western blot鉴定目的基因表达,利用MicroRNA芯片技术检测其对L02细胞MicroRNA表达的影响。结果 RT-PCR及Western blot表明在L02/HBx和L02/HBx-d382细胞株中存在目的基因表达。MicroRNA芯片发现与L02细胞相比,L02/HBx-d382细胞有7个MicroRNA表达上调,5个MicroRNA表达下调,L02/HBx细胞有4个MicroRNA表达上调,12个MicroRNA表达下调。结论成功构建HBx基因及其缺失突变体(HBx-d382)的真核表达模型,该类病毒基因会影响L02肝细胞的MicroRNA表达谱。ObjectiveTo study the microRNA expression profile in L02 cells transfected with HBx gene or its deletion mutation（HBx-d382）.MethodsPositive clones of the L02/HBx and L02/HBx-d382 cells were obtained by liposome transfection and G418 selection.Expression of the HBx or HBx-d382 was confirmed by reverse transcription PCR（RT-PCR） and Western blot.MicroRNA expression profiles were analyzed by MicroRNA microarray.ResultsRT-PCR and Western blot showed that L02/HBx and L02/HBx-d382 cells had target gene expression.Based on the microRNA microarray,7 up-regulated and 5 down-regulated microRNAs were observed in L02/HBxd382 cells;and 4 up-regulated and 12 down-regulated microRNAs were observed in L02/HBx,compared with L02 cells.ConclusionL02 cell line stably expressing HBx or HBx deletion mutation is successfully established.This virus gene can influence the microRNA expression profile in L02 cells.

关 键 词：HBX 突变 微小RNA 微小RNA芯片 

分 类 号：R735.7[医药卫生—肿瘤]