Hepatocellular carcinoma (HCC) is one of the most common solid tumors worldwide. Epigenetic changes in gene expression, including DNA methylation and histone modifications, may contribute to the development of HCC. Polymorphisms of the DNA methyltransferase 3B (DNMT3B) gene may affect the activity of this enzyme and increase the susceptibility to several types of cancer, including HCC. To confirm this hypothesis, we investigated the association between single-nucleotide polymorphisms-149C>T (rs2424913) and -579G>T (rs1569686) in the promoter region of DNMT3B and the risk of HCC. DNMT single-nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction-restriction fragment length polymorphism in 108 HCC patients and 240 healthy controls matched for age, gender and ethnicity. The DNMT3B-149 TT genotype was not significantly associated with an increased risk of HCC. The frequency of DNMT3B-149C was 0.46% in HCC patients and 1.39% in healthy individuals, whereas the frequency of DNMT3B-579G was 8.33% in HCC patients and 10.42% in healthy individuals. No significant differences were observed in the genotype or allelic distribution between HCC patients and controls. In conclusion, DNMT3B-149C>T and -579G>T polymorphisms are not significantly associated with an increased risk of HCC. These results demonstrated that these particular SNPs may not be used as biomarkers to predict susceptibility to HCC.