The stimulator of interferon genes (STING) plays a crucial role in the recognition of a viral infection and subsequent stimulation of an immune response. However, it is unclear whether methylation of the STING promoter affects STING transcription and response to antiviral therapy. The present study determined the methylation status of the STING promoter in patients with chronic hepatitis B (CHB).This study included 198 participants, of which 159 participants had CHB and 39 were healthy controls (HCs). Methylation-specific polymerase chain reaction was performed to detect the methylation status of the STING promoter. Reverse transcription-quantitative polymerase chain reaction was performed to determine STING mRNA level in peripheral blood mononuclear cells.The methylation frequency of the STING promoter was significantly higher and STING mRNA level was lower in the patients with CHB than in the HCs. Presence of hepatitis B virus (HBV) DNA was independently correlated with an increased risk of STING promoter methylation. Virological response frequency was higher in the patients with CHB receiving entecavir (ETV) than in those receiving adefovir (ADV). In the ETV group, the virological response frequency was evidently lower in the patients with CHB having methylated STING promoters than in those having unmethylated STING promoters. However, there was no significant difference in the virological response frequency between ADV-treated patients having methylated and unmethylated STING promoters.These results indicate that the hypermethylation of the STING promoter and thus the transcriptional repression of STING weaken the effect of STING in inhibiting HBV replication and decreases the effectiveness of antiviral therapy.