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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。
Objective To investigate whether hepatitis C virus (HCV) core protein affects the expression of cyclooxygenase-2 (COX-2). Methods The HCV core protein gene was amplified by PCR from the full-length HCVH77-containing plasmid and cloned into pcDNA3.1 vector to construct HCV core protein HCV-C / pcDNA3.1. The HepG2 cells were co-transfected with the luciferase reporter vector COX2pro1.5kb / luc containing the COX-2 promoter and HCV-C / pcDNA3.1. The firefly luciferase activity was measured and the expression of COX-2 protein was detected by Western blot Level. Results The HCV-C / pcD-NA3.1 recombinant plasmid was successfully constructed. In transiently transfected HepG2 cells, the COX2 promoter luciferase activity was significantly enhanced. Western blot analysis showed that the expression of COX-2 was significantly increased. Conclusion HCV core protein activates COX-2 promoter in HepG2 cells and obviously induces the expression of COX-2, which provides a new experimental basis for further study on the relationship between COX-2 and the pathogenesis of HCV.