miR-122对乙型肝炎病毒抗原表达的影响

摘要

Objective; To investigate the effect of miR-122 on the expression of hepatitis B virus (HBV) proteins. Methods: Anti-sense oligodeoxynucleotide ( ASODN) of two different sequences against miR-122, anti-miR-122 and LNA-antimiR-122 (Locked nucleic acid), human miR-122 ( has -miR-122), or the negative control anti-GFP were designed and synthesized, then transfected into HepG2. 2.15 cells. After 24 h and 48 h, the levels of HBsAg and HBeAg in the supernatant were determined with a time-resolved immunofluorometric assay (TRFIA). HBV DNA in supernatant and miR -122 in cells were measured by quantitative real-time PCR. Results; After 48 h expressions of miR-122 in the LNA-antimiR-122 and anti-miR-122 groups were significantly suppressed and lower than those in the negative control (P <0.001), while the level of miR-122 in the has-miR-122 group was higher than that in the negative control (P <0. 001). The expression of HBeAg and HBsAg in has-miR-122 group was lower than that in the negative control (P<0.01) 24 h and 48 h after transfection. The expression of HBeAg and HBsAg in the anti-miR-122 group and LNA-antimiR-122 group was significantly lower than that in negative control (P < 0. 001). The levels of viral DNA at both time-points in the various test groups were not significantly different from those of negative control group (P >0. 05). Conclusion; miR-122 may regulate HBV antigens and potentially affect the progress of pathogenesis, which might be the new targets for treatment of HBV infection.%目的:探讨miR-122对乙型肝炎病毒HBsAg、HBeAg表达的影响.方法:设计合成2务miR-122的反义寡核苷酸(anti-miR-122,LNA-antimiR-122)、hsa-miR-122以及阴性对照anti-GFP,脂质体介导转染HepG2.2.15细胞.取细胞转染24h、48 h后的培养液,时间分辨荧光免疫法检测anti-miR-122组、LNA-antimiR-122组、hsa-miR-122组以及anti-GFP组HBsAg和HBeAg的表达,定量PCR检测各组HBV DNA的表达.转染48 h后,提取细胞内总RNA,Taqman技术实时荧光定量PCR检测各组miR-122的表达.结果:①转染48 h后anti-miR-122组、LNA-antimiR-122组miR-122表达明显受抑制,低于阴性对照组(P ＜0.001).hsa-miR-122组miR-122水平较阴性对照组升高(P＜0.001).②hsa-miR-122组HBsAg、HBeAg表达均低于阴性对照组(P ＜0.001)；anti-miR-122组、LNA-antimiR-122组HBsAg、HBeAg表达均高于阴性对照组(P ＜0.001).③转染24 h与48 h后各组HBV DNA表达与阴性对照组比较无统计学意义(P＞0.05).结论:miR-122可调节乙型肝炎病毒抗原表达.