HBx基因缺失突变体(HBx-d382)对L02细胞cyclinD1、cyclinG1和E2F1表达的影响  

Influence of HBx gene deletion mutation(HBx-d382) on cyclin D1,cyclinG1 and E2F1 expression in L02 cells

作　　者：胡志亮[1] 侯周华[1] 符小玉[1] 陈莉[1] 谢萍[1] 欧阳奕[1] 杨永峰[2] 谭德明[1] 

机构地区：[1]中南大学湘雅医院感染科,湖南长沙410008 [2]东南大学附属南京市第二医院感染科,江苏南京210003

出　　处：《中国现代医学杂志》2011年第33期4114-4117,4121,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(No:30872228);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：目的前期的研究成功构建了稳定表达HBx基因及其缺失突变体(HBx-d382)的L02细胞,分别命名为L02/HBx和L02/HBx-d382,并证实其可导致肝细胞恶性转化。该研究进一步探讨HBx-d382对L02细胞G1期细胞周期调控相关基因cyclinD1,cyclinG1和E2F1表达的影响。方法实验分为L02/pcDNA3.0(稳定转染pcDNA3.0)、L02/HBx和L02/HBx-d382(分别稳定转染质粒pcDNA3.0/HBx和pcDNA3.0/HBx-d382)3组。通过实时定量PCR以及wester-blot检测转染HBx基因及其缺失突变体HBx-d382后L02细胞cyclinD1、cyclinG1和E2F1表达的改变。方法实时定量PCR以及wester-blot结果表明稳定表达HBx基因或HBx-d382的L02细胞cyclinD1、cyclinG1和E2F1表达上调,表达HBx-d382的L02细胞上调最为明显。结论 HBx-d382可能通过上调cyclinD1、cyclinG1和E2F1从而影响细胞G1期调控,进一步导致肝细胞恶性转化。[Objective] Previously we had established L02 cell lines stably expressing HBx gene or its deletion mutant （HBx-d382）, which were shown to be implicated in hepatoeytes malignant transformation, named L02/HBx and L02/HBx-d382 respectively. This study further investigated the influence of HBx-d382 on cyclin D1, cyelin G1 and E2F1 expression associated with G1 cell cycle control in I1）2 cells. [ Methods] Cell lines LO2/pcDNA3, L02/ HBx and L02/HBx-d382 （stably transfeeted with plasmid pcDNA3.0, pcDNA3.0/HBx and pcDNA3.0/HBx-d382, respectively） were evaluated in this study. The mRNA and protein expression of eyclin D1, cyclin G1, E2F1 were detected by Real-time Quantitative PCR and Western blot respectively. [ Results ] Real-time Quantitative PCR and Western blot showed that the expression of cyclin D1, cyclin G1 and E2F1 were up-regulated in L02/HBx and L02/ HBx-d382 cell lines ,which was more significant in L02/HBx-d382 group. [ Conclusion ] HBx-d382 up-regulates the expression of cyclin D1, cyclin G1 and E2F1 in L02 cells which influences the G1 cell cycle control and may further contribute to liver carcinogenesis.

关 键 词：HBX HBx-d382 细胞周期 CYCLIN 

分 类 号：R361.2[医药卫生—病理学]