摘要
目的:研究乙型肝炎病毒复制子pHY106-BHBV转染肝癌细胞系基因表达谱的差异,分析参与胆固醇代谢的差异表达基因.方法:应用包含20000条人类全长基因的寡聚核苷酸芯片ImaGene3.0检测乙型肝炎病毒复制子pHY106-BHBV转染肝癌细胞系基因表达谱差异,并选择其中参与胆固醇代谢的表达下调基因7-脱氢胆固醇还原酶和NADH-细胞色素b5还原酶,用RT-PCR方法进行验证分析.结果:基因芯片筛选出差异表达的基因,其中包括转录因子、细胞周期素、细胞因子相关蛋白、糖脂类物质代谢相关酶及细胞信号转导相关因子等,RT-PCR验证参与胆固醇代谢的几种表达差异的基因,其中7-脱氢胆固醇还原酶和NADH-细胞色素b5还原酶表达明显下调,与基因芯片分析结果一致.结论:乙型肝炎病毒复制子pHY106-BHBV转染肝癌细胞系存在基因表达差异,乙型肝炎病毒感染可能通过抑制参与胆固醇代谢的7-脱氢胆固醇还原酶和NADH-细胞色素b5还原酶从而进一步影响机体胆固醇代谢.
AIM：To investigate the changes in gene expression profile of HepG2 cells transfected with HBV replicon pHY106-BHBV by cDNA microarray,and to identify differential genes involved in cholesterol metabolism.METHODS：A cDNA microarray （Affymetrix） that contains 20 000 gene sequences was used to examine gene expression between HepG2 cells transfected with HBV replicon pHY106-BHBV and control plasmid.Differentially expressed genes that may contribute to cholesterol metabolism were subjected to real-time reverse transcription-polymerase chain reaction （RT-PCR） analysis.RESULTS：The differentially expressed genesidentified mostly encode transcription factors,cytokines,and molecules that are involved in signal transduction,glucose metabolism and lipid metabolism.Real-time RT-PCR results indicated that cholesterol metabolism-related genes 7-dehydrocholesterol reductase （dhcr7） and NADH-cytochrome b5 reductase （cyb5r） were down-regulated in HepG2 cells transfected with HBV replicon pHY106-BHBV.CONCLUSION：Differential gene expression profile is detected between HepG2 cells transfected with HBV replicon pHY106-BHBV and control plasmid.The expression of cholesterol metabolism-related genes dhcr7 and cyb5r may be suppressed by HBV infection.
出处
《世界华人消化杂志》
CAS
北大核心
2010年第10期1041-1045,共5页
World Chinese Journal of Digestology
