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目的探讨干扰素β启动子刺激因子1(IPS-1)不同单核苷酸多态性(SNP)基因型对HBV感染后干扰素应答作用机制。方法利用IPS-1野生型和rs17857295、rs7262903、rs7269320三种SNP基因型慢病毒表达载体,转染目的细胞HepG2、HepG2.2.15细胞,qPCR检测HBV感染和IPS-1不同SNP基因型对目的基因表达情况的影响,检测细胞转染后IFN-βmRNA表达水平,探讨IPS-1不同SNP基因型对HBV感染后细胞干扰素应答的机制。结果 IPS-1rs17857295和rs7262903两种SNP基因型转染HepG2.2.15细胞后,IPS-1基因表达水平显著低于野生型转染组(451.77比712.53,498.71比712.53,P<0.01);但IFN-βmRNA表达水平显著高于野生型转染组(分别为5.33比0.98和3.59比0.98,P<0.01);rs7269320 SNP基因型过表达后,IPS-1基因表达水平显著低于野生型转染组(290.63比712.53,P<0.01),但IFN-βmRNA表达水平无明显变化(0.74比0.98)。另外,rs17859295基因型分别转染HepG2和HepG2.2.15细胞后,后者IFN-βmRNA表达水平显著高于前者(5.33比2.25,P<0.01)。结论 IPS-1 rs17857295和rs7262903两种SNP基因型HBV感染者,尤其rs17857295 SNP基因型,清除病毒的能力可能强于野生型的感染者;而IPS-1野生型和rs7269320 SNP基因型HBV感染者可能更易形成病毒感染的慢性化。
Objective To investigate the mechanism of IFN-β response to hepatitis B virus infection by different single nucleotide polymorphisms (SNPs) of interferon β promoter stimulator 1 (IPS-1). Methods IPS-1 wild-type and rs17857295, rs7262903 and rs7269320 lentiviral expression vectors were transfected into HepG2 and HepG2.2.15 cells. QPCR was used to detect the expression of target gene in HBV infection and IPS-1 SNP The effect of different SNP genotypes of IPS-1 on the IFN-βmRNA expression level after transfection was examined. Results The expression of IPS-1 gene in HepG2.2.15 cells transfected with two SNP genotypes IPS-1rs17857295 and rs7262903 was significantly lower than that in wild-type transfected cells (451.77 vs 712.53, 498.71 vs 712.53, P <0.01) β mRNA was significantly higher than that of wild-type group (5.33 vs 0.98 and 3.59 vs 0.98, respectively, P <0.01). The over-expression of rs7269320 SNP gene was significantly lower than that of wild-type group 290.63 vs 712.53, P <0.01), but the level of IFN-βmRNA did not change significantly (0.74 vs 0.98). In addition, rs17859295 genotype transfected HepG2 and HepG2.2.15 cells, the latter IFN-βmRNA expression was significantly higher than the former (5.33 vs 2.25, P <0.01). Conclusion The two SNP genotypes of rs17857295 and rs7262903 in IPS-1 infected patients, especially rs17857295 SNP genotype, may be able to clear the virus more than wild-type ones. However, both the IPS-1 wild-type and rs7269320 SNP genotype HBV carriers may Easier to form a chronic viral infection.