HCV核心蛋白在HepG2细胞中对COX-2表达的影响

The effect of HCV core protein on the expression of cyclooxygenase 2 (COX-2) in HepG2 cells

目的研究丙型肝炎病毒(HCV)核心蛋白是否影响环氧化酶-2(COX-2)的表达。方法用PCR从含HCVH77株全长基因组序列质粒中扩增HCV核心蛋白基因,并克隆至pcDNA3.1载体中,构建HCV核心蛋白基因的真核表达载体HCV-C/pcDNA3.1。用HCV-C/pcDNA3.1和含COX-2启动子的荧光素酶报告载体COX2pro1.5kb/luc瞬时共转染HepG2细胞,测定萤火虫荧光素酶的活性,并用Westernblot检测COX-2蛋白的表达水平。结果成功地构建了HCV-C/pcD-NA3.1重组质粒。瞬时转染后的HepG2细胞中,COX2启动子荧光素酶的活性显著增强。Westernblot检测,发现COX-2的表达明显升高。结论HCV核心蛋白在HepG2细胞中激活COX-2启动子,并且明显诱导COX-2的表达,为进一步研究COX-2与HCV致病性的关系提供了新的实验依据。

AIM： To investigate the effect of Hepatitis C virus （HCV） core protein on the expression of cyclooxygenase 2 （COX-2）. METHODS： The genes encoded HCV core protein were amplified from plasmid containing full length genome of HCV strain 1-177 using PCR, and were cloned into eukaryotic expression vector pcDNA3. 1, The recombinant HCV-C/pcDNA3. 1 was transiently co-transfected into HepG2 cells with luciferase reporter vector containing COX-2 promotor （COX2pro1. 5 kb/luc）, The luciferase activity and COX-2 protein expression were detected. RESULTS： The reombinants HCV-C/pcDNA3.1 have been constructed successfully. The luciferase activity of COX-2 promotor was activated by the expressed HCV core, and the increased protein expression of COX-2 in transfected HepG2 cells was detected by Westem blot. CONCLUSION： HCV core protein can activate the COX-2 promotor and induce its expression, which provides a new experimental basis for further reseach on relationship between COX-2 and HCV pathogenesis.