HepG2.2.15细胞系中HBV复制相关抑制因素的变化

Changes of HBV replication-associated inhibitory factors in HepG2.2.15 cells

目的:研究HBV对其自身复制相关抑制因素的影响,为今后探索提高抗病毒治疗的疗效提供基础。方法:采用实时荧光定量RT-PCR技术检测原蛋白转化酶(PCs)、肝细胞核因子(HNF)3β和转化生长因子(TGF)β1等HBV复制相关抑制因素的mRNA表达,采用Western印迹技术检测部分PCs的蛋白表达,通过比较模型细胞HepG2.2.15和其前体细胞HepG2的表达差别来判断HBV对其自身复制相关抑制因素的影响。结果:HepG2.2.15和HepG2细胞均相对高表达furin和PACE4,但HepG2细胞还高表达PC1/2。与HepG2细胞相比,HepG2.2.15细胞的PC1/3和PC7/8在mRNA水平显著被下调(均P<0.05),且PC1/3在蛋白水平的表达也显著减少。HepG2.2.15细胞的HNF3β和TGFβ1的mRNA表达水平也发生了显著改变,分别增加到5.22倍和降低到0.35倍。结论:HBV的存在对其自身复制相关抑制因素可产生显著影响。下调部分PCs和TGFβ1表达可能有利于HBV自身复制,但上调HNF3β的意义值得深入研究。

Objective To investigate the effects of HBV on its replication-associated inhibitory factors, and to provide information to promote antiviral efficacy in the future. Methods Expressions of proprotein convertases （PCs）, hepatoeyte nuclear factor （HNF） 3β, and transforming-growth factor （TGF） β1 mRNA were detected by quantitative real-time RT-PCR technique and protein of some PCs were detected by western blot analysis. The effects of HBV on these replication-associated inhibitory factors were obtained by comparing the difference between HepG2.2.15 cell line and its precursor HepG2 cell line. Results Furin and PACE4 mRNA were highly expressed in both HepG2.2.15 and HepG2 cells, and PC1/3 was highly expressed in HepG2 cells. As compared with those in HepG2 cells, PC1/3 and PC7/8 mRNA were down-regulated in HepG2.2.15 cells （P 〈 0.05）, and PC1/3 protein was also down-regulated. HNF313 and TGFβ1 mRNA in HepG2.2.15 cells were up-regulated to 5.22 times and down-regulated to 0.35 times, respectively. Conclusions HBV exerts significant effects on its replication-associated inhibitory factors. The down-regulated expressions of some PCs and TGFβ1 might be favorable for HBV replication, but the importance of the up-regulated expression of HNF3β urges for more studies in the future.