【摘要】 目的探讨丙型肝炎病毒(HCV)核心蛋白对转录因子ETF和TxRE活性的调控作用。方法对于瞬时转染p IRES-Core/Neo载体的L02细胞采用双荧光素酶报告基因系统检测核心蛋白对转录因子ETF和TxRE活性影响,并提取2株细胞的核蛋白和RNA,分别以Western blot试验和实时荧光定量PCR检测2种转录因子的表达量。结果双荧光素酶报告基因检测在L02-Core和L02-Neo细胞中转录因子ETF和TxRE的活性比值分别为2.981和3.063。Western blot试验灰度值分析2株细胞的核蛋白提取物中这2种蛋白表达量的比值分别为1.510和2.717。实时荧光定量PCR检测这2株细胞中ETF和TxRE的核酸水平比值分别为4.123和3.703。结论在表达HCV核心蛋白的L02细胞中,转录因子ETF和TxRE转录表达水平及转录活性均较高,HCV核心蛋白可能具有增强转录因子ETF和TxRE转录活性的作用。更多还原

【Abstract】 Objective To investigate the regulation of the hepatitis C virus( HCV) core protein on the activities of transcription factors ETF and TxRE. Methods The effect of HCV core protein on the activities of transcription factors ETF and TxRE was detected by dual luciferase reporter gene system and Western blot experiments. Real-time quantitative PCR was applied to detect the expression of the two transcription factors by transient transfection with p IRES-Core / Neo vector in L02 cells. Results The dual luciferase reporter gene assay results showed that the ratios of activities of ETF and TxRE were approximately 2. 981 and 3. 063 in the L02-Core and L02-Neo cells. The results of Western blot showed that the expression of ETF and TxRE in L02-Core was higher than that in L02-Neo and the ratios were about 1. 510 and 2. 717. Real-time PCR results showed that the mRNA ratios of ETF and TxRE in the two cell lines were around 4. 123 and 3. 703,respectively. Conclusion The transcription expression and activity of transcription factors ETF and TxRE in L02 cells expressing HCV core protein were higher than those in the corresponding control cell. Thus,the hepatitis C virus core protein can enhance the transcriptional activity of ETF and TxRE in the human liver cell line L02.更多还原