目的 探讨HCV核心蛋白对低氧诱导因子1α【(HIF-1α)和血管内皮生长因子(VEGF)表达的影响。方法将HCV核心蛋白基因的真核表达载体flag2B—core和HIF-1αsiRNA转染Huh7.5.1细胞;采用RT-PCR和Westernblot法分别检0n,0HIF~1α、VEGF在mRNA和蛋白水平的表达变化,采用酶联免疫吸附试验检测细胞上清液中VEGF的含量。对各组间数据进行t检验。结果Huh7.5.1细胞转染flag2B-core后,HIF-1α和VEGF的mRNA和蛋白表达水平升高,细胞上清液中VEGF含量明显高于对照组[(654.5±43.7)pg/ml与(365.9±26.8)pg/ml,t=653.1%,P〈0.01)];Huh7.5.1细胞共转染flag2B—core和HIF-1asiRNA后,HIF-1a和VEGF的mRNA和蛋白表达水平降低,细胞上清液VEGF含量明显低于对照组[(389.2±29.6)pg/ml与(768.8±47.3)pg/ml,t=1330.22,P〈0.01)。结论HCV核心蛋白能够上调HIE-1α和VEGF的表达;HCV可能通过核心蛋白来调节HIF-1α和VEGF的表达。
Objective To investigate the effect of core protein of hepatitis C virus (HCV) on the expression of hypoxia-inducible factor-1α (HIF-1α and vascular endothelial growth factor (VEGF). Methods HuhT.5.1 cells were transfected with plasmid flag2B-core carrying HCV core gene, expression of HIF-1α and VEGF were measured by reverse transcription-polymorphism chain reaction (RT-PCR) and western blot. Enzyme link immunobsorbent assay (ELISA) were used to detect the level of VEGF in the supernatants. Results The expression of HIF-1α and VEGF mRNA and protein were upregulated after flag2B-core was transfected into HuhT.5.1 cells, and VEGF level in the supematant was significant elevated as compared to controls [(654.5±43.7) pg/ml vs (365.9±26.8) pg/ml, t = 653.1%, P〈0.01]. The expression of HIF-1a and VEGF mRNA and protein were downregulated after flag2B-core and HIF-1a siRNA were co-transfected into Huh7.5.1 cells, and VEGF level in the supernatant was significanty reduced as compared to controls [(389.2±29.6) pg/ml vs (768.8±47.3)pg/ml, t = 1330.22, P〈0.01]. Conclusions HCV core protein enhances the expression of HIF-1a and VEGF. HCV may regulate the expression of HIF-1a and VEGF via the core protein.