Association Of Hepatocellular Carcinoma With S-gene Additional N-glycosylation Mutation Of Hepatitis B Virus In HBs Ag And Anti-HBs Coexistent Patients

Objective To investigate the association of additional N-glycosylation mutation in major hydrophilic region(MHR)of hepatitis B virus(HBV)S gene with the risk of hepatocellular carcinoma(HCC)in HBsAg and anti-HBs coexistent patients.Methods A total of 288 HBsAg/anti-HBs coexistent patients and 490 single HBsAg-positive patients were enrolled,who were admitted in 302 Hospital of PLA.from July 2009 to June 2016.HBV DNA was extracted from serum samples and subjected to nested PCR for full-length S-gene sequencing.The association of MHR additional N-glycosylation mutation and clinical parameters with HCC occurrence risk was analyzed.Specifically,the additional N-glycosylation mutation was dynamically analyzed prior and post HCC occurrence for 18 patients.Results The frequency escalated by an order of single HBsAg-positive non-HCC(1.6%),single HBsAg-positive HCC(6.0%),HBsAg/anti-HBs coexistent non-HCC(8.0%),and HBsAg/anti-HBs coexistent HCC(20.8%).Twelve kinds of mutations/mutation patterns were detected,five of which have not been reported.Multivariate analysis showed that Age >40 years,HBsAg >median,HBeAg negativity,and additional N-glycosylation mutation in MHR were associated with HCC risk for the HBsAg and anti-HBs coexistent patients(OR = 4.281,95%CI 1.843-9.941,P = 0.001;OR = 3.146,95% CI 1.633-6.060,P = 0.001;OR = 2.097,95% CI 1.010-4.357,P = 0.047;and OR = 4.381,95% CI 1.842-10.417,P = 0.001).In contrast,ALT,anti-HBs,anti-HBe,and HBV DNA levels had no significant association with HCC occurrence.Dynamical analysis showed that the additional N-glycosylation mutation already developed 1-4 years prior HCC occurrence in the eight of 18 observed patients.Conclusion Additional N-glycosylation mutation in MHR of HBV S gene had close association with HCC occurrence in HBsAg and anti-HBs coexistent patients,suggesting that HBsAg and anti-HBs coexsistence and the additional N-glycosylation mutation together could serve as a predictive indicator for HCC occurrence risk in chronic HBV-infected patients.