Effects Of MiR-122 On Antigen Expression Of Hepatitis B Virus

Objective To investigate the effect of miR-122 on the expression of corresponding protein of HBV:HBsAg and HBeAg.Methods miR-122 antisense oligodeoxynucleotide(ASODN)of two different sequences anti-miR-122 and LNA-antimiR-122(Locked nucleic acid-anti-miR-122 GFP;hsa-miR-122(human miR-122),GFP(GFP antisense oligodeoxynucleotide)were designed and synthesized.Anti-miR-122,GFP,LNA-antimiR-122,hsa-miR-122 with the final concentration of 400nm/L,400nm/L,40nm/L,400nm/L were delivered into HepG2.2.15 cells by Oligofectamine^TMreagent,respectively.GFP group and normal control group were set as negative control.After 24h and 48h,the levels of HBsAg and HBeAg expression in the supernatant were determined with Timeresolved Immunofluorom-etric Assay kit(TRFIA),respectively.After 48h,the total RNA of all cells was extracted,and here we developed and evaluated a rapid and reliable real-time quantitative reverse transcription polymerase chain reaction(real time RT-PCR)approach using TaqMan(?)MicroRNA Assays technology for detection of the expression level of miR-122.Results①After 24 h,the expression level of HBsAg:The expression level of HBsAg in hsa-miR-122 group was lower than that in negative control(p＜0.05)and that in anti-miR-122group(p＜0.01).The expression level of HBsAg in anti-miR-122 was higher than that in GFP group(p＜0.01).②After 24 h,the expression level of HBeAg:The expression level of HBeAg in anti-miR-122 group was higher than that in negative control(p＜0.01),the expression level of HBeAg in LNA-antimiR-122 group was higher than that in negative control(p＜0.01),the expression level of HBeAg in hsa-miR-122 group was lower than that in anti-miR-122 group(p＜0.01),the expression level of HBeAg in GFP group is lower than that in anti-miR-122 group(p＜0.01).③After 48h,the expression level of HBsAg:The expression level of HBsAg in GFP group is equal to that in negative control(p=1.000),the expression level of HBsAg in anti-miR-122 group is higher than that in negative control(p＜0.05)and that in GFP group(p＜0.05),the expression level of HBsAg in hsa-miR-122 group was lower than that in anti-miR-122 group(p＜0.01) and that in LNA-antimiR-122 group(p＜0.05).④After 48h,the expression level of HBeAg:The expression level of HBeAg in LNA-antimiR-122 group was higher than that in negative control(p＜0.05),the expression level of HBeAg in hsa-miR-122 group was lower than that in LNA-antimiR-122 group(p＜0.05),the expression level of HBeAg in GFP group is lower than that in LNA-antimiR-122 group(p＜0.05).⑤The expression level of miR-122:After 48h,The results of real time RT—PCR demonstrated that the expression level of miR-122 in LNA-antimiR-122 group and anti-miR-122 group was lower than that in negative control,the expression level of miR-122 in hsa-miR-122 group was higher than that in negative control,the expression level of miR-122 in GFP group was equal to that in negative control. Conclusion These results show that there is an inverse correlation between level of miR-122 and HBsAg.When miR-122 of HepG2.2.15 was depressed,the expression of HBeAg was up-regulated prominently.