Epidemiologic studies have suggested that cigarette smoking is a risk factor for the development of hepatocellular carcinoma (HCC). To further investigate this relation, the authors measured levels of 4-aminobiphenyl-DNA adducts by an immunoperoxidase method in surgical liver tissues obtained between 1984 and 1995 from 105 Taiwanese patients with histologically confirmed HCC and 37 Taiwanese patients with metastatic liver tumors or intrahepatic stones. Information on clinicopathologic characteristics, cigarette smoking, and alcohol drinking was abstracted from hospital charts. Mean relative staining intensity for 4-aminobiphenyl-DNA was slightly higher in tumor tissues than in nontumor tissues obtained from HCC patients. Both mean intensities were significantly higher than the mean intensity of control tissues taken from non-HCC patients. However, no difference in mean relative staining intensity was found between smokers and nonsmokers in tissues obtained from non-HCC patients, or in tumor or nontumor tissues taken from HCC cases. After stratification of the relative staining intensities of 4-aminobiphenyl-DNA adduct levels into tertiles according to the total numbers of control tissues analyzed, there was a monotonically increasing risk of HCC. Odds ratios were 4.14 (95% confidence interval (CI) 1.15-15.50) and 9.71 (95% CI 2.82-34.86) for medium and high adduct levels compared with low adduct levels, respectively. The linear relation between adduct levels in liver tissue and HCC risk was also significant after adjustment for covariates, including hepatitis B surface antigen (HBsAg) status. The multivariate adjusted odds ratios were 3.41 (95% CI 0.82-14.25) and 6.48 (95% CI 1.59-26.50) for medium and high adduct levels, respectively. Moreover, there were monotonically increasing HCC risks for higher adduct levels in both HBsAg carriers and noncarriers. The increased risk ratios were more pronounced in noncarriers than in carriers. However, because of the small numbers of subjects, especially controls positive for HBsAg, the interaction between HBsAg status and 4-aminobiphenyl-DNA adduct level was not significant. Among HCC cases, none of the clinicopathologic characteristics were associated with relative staining intensity. These results indicate that 4-aminobiphenyl exposure, which is primarily a result of cigarette smoking, plays a role in the development of HCC in humans.