The aims of this study were to evaluate the efficacy of long‐term interferon (IFN) monotherapy on hepatocellular carcinoma (HCC) in patients who showed no virological response to the first course of IFN therapy, define predictive factors for HCC in patients on long‐term IFN monotherapy, and evaluate the clinical impact of amino acid (aa) substitutions in the hepatitis C virus (HCV)‐1b core region on HCC rate. This retrospective study included 494 consecutive treatment‐naive patients infected with HCV‐1b who failed to achieve sustained virological response after ≥24‐week IFN monotherapy. Of 494 patients, 113 (22.9%) received another course of ≥48‐week IFN monotherapy (additional‐IFN group), while the remaining 381 (77.1%) received no such therapy (no‐additional‐IFN group), and 10 years have elapsed since the end of the first IFN monotherapy. The cumulative HCC rate was significantly higher in the no‐additional‐IFN group than additional‐IFN group, and in those with aa substitutions in the core region of Gln70(His 70) and Met 91 than those with Arg 70 and/or Leu 91. Multivariate analysis identified stage of liver fibrosis, liver enzymes, age, treatment group, aa substitution in the core region, low‐density lipoprotein cholesterol (LDL‐cholesterol), and gender as determinants of HCC, and that additional IFN treatment significantly lowered the cumulative rate of HCC, even in patients with cirrhosis. In conclusion, long‐term IFN monotherapy reduces the risk of HCC, even in patients with cirrhosis. Substitution of aa at position 70 and/or 91 in the core region and lipid metabolism are important predictors of HCC in long‐term IFN monotherapy. J. Med. Virol. 84: 1199–1207, 2012. © 2012 Wiley Periodicals, Inc.