CYP1B1 is a P450 enzyme which is involved in the activation of pro-carcinogens to carcinogens as well as estrogen metabolism. We hypothesized that genetic variants in CYP1B1 may modify individual susceptibility to hepatocellular carcinoma (HCC).To test this hypothesis, we evaluated the associations of three CYP1B1 single nucleotide polymorphisms (SNPs) and HCC risk in a case-control study of 468 HCC cases and 515 cancer-free controls in a Chinese population. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry method and direct DNA sequencing were performed to detect these polymorphisms.In overall analysis, we found that only the variant G allele of rs1056836 was associated with a significantly increased risk of HCC among the three SNPs (rs10012, rs1056836 and rs1800440). Moreover, we found that the variant genotypes containing the G allele of rs1056836 were associated with a significantly increased risk of HCC among HbsAg-positive individuals (adjusted OR=2.13, 95% CI=1.18, 3.86), but not among HbsAg-negative individuals. When stratifying by smoking status, we found that the variant GG genotype increased a 13.97-fold (95% CI=1.28, 152.94) risk of HCC among smokers. Furthermore, high risk for liver cirrhosis-positive clinical status was exhibited in HCC patients with rs1056836 CG and GG genotypes as compared with CC homozygotes. For the other two SNPs, we did not find any significant evidence of association with HCC risk in any subgroup.This study suggests that CYP1B1 rs1056836 polymorphism may be an important factor contributing to increased susceptibility and pathological development of HCC in Chinese population.