The Roles Of ADAR1 In The Anti-HBV Mechanisms Of Interferon

Objective: To investigate the roles of ADAR1 in the anti-HBV mechanisms of interferon in HCC cells.Methods:1.RT-qPCR and Western blot were used to detect the expression of ADAR1 in HepG2 cell lines treated with different concentrations of interferon,type ? IFN(IFN-?: 0,200,500,1000,2000 U/ml and IFN-?: 0,200,1000 U/ml)and type ? IFN(IFN-?: 0,200,1000 U/ml)under different time(0,2,4,8,16h)of IFN-? treatment.2.Huh7 cell lines were transfected with HBV replication plasmids then treated with IFN-?.Southern blot analysis was carried out to detect the changes in HBV DNA level while ELISA was employed to detect the expression of HBsAg levels.3.The expression of ADAR1 in Huh7 cell lines transfected with HBV replication plasmids was knocked down by siRNA followed by IFN-?,Southern blot was used to detect the changes in HBV DNA level while Western blot was used to detect the expression of ADAR1.4.The change of genes expression related to ADAR1 wereinvestigated and verified in Huh7.5.1 cells prior and post ADAR1-knockout through transcriptome sequencing analysis.Results:1.mRNA levels of ADAR1(P110 and P150)were increased when treated with type ? interferon(IFN-?,IFN-?),and the increase was IFN-?dose-dependent,type ? interferon(IFN-?)did not significantly affected the expression of ADAR1 in transcription level.2.The protein levels of ADAR1 including P110 and P150 were remarkably increased after the treatment of type ? interferon(IFN-?,IFN-?),and the expression was IFN-? time-dependent.Type ? interferon(IFN-?)did not significantly affected the expression of ADAR1 in protein level.3.ADAR1 may play a role in this process of IFN-? treatment by replication inhibition of HBV DNA in Huh7 HCC cells.4.Based on the RNA-seq data,2946 differential expression genes and related signaling pathways were screened in Huh7.5.1 cells and Huh7.5.1g cells and 25 hub genes screened were verified.Conclusion:ADAR1 expression on protein and mRNA levels were stimulated by type ? interferon.And ADAR1 may be involved in the inhibition of HBV replication by interfion.