miR-122对乙型肝炎病毒抗原表达的影响

Effects of miR-122 on expression of hepatitis B virus proteins

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条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24 h、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。

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(hsa-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 hsa-miR-122 group was higher than that in the negative control(P<0.001).The expression of HBeAg and HBsAg in hsa-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.