Our study has limitations. First, we failed to amplify Navitoclax supplier the two HBV fragments from all of HBV-infected population, resulting in a possible preponderance of missing data. Failure to amplify the HBV region might be due to low viral concentration in sera or HBV mutations in the primer binding sites. Second, other environmental exposures such as alcohol consumption and family history of cancer were incomplete and thus not included in the analyses. Third, healthy controls were selected from one of the hospitals from which cases were recruited. This may cause potential selection bias. Fourth, the study design is cross-sectional in nature. Future prospective studies are needed to confirm the synergistic or antagonistic effect of the SNPs with the HBV mutations in hepatocarcinogenesis and define the HBV-infected subjects who are more likely to develop HCC and need specific interventions.

In conclusion, our study suggested that pri-miR-34b/c rs4938723 was associated with a significant increased risk of HCC, especially in women. Although pre-miR-196a2 rs11614913 was not statistically associated with HCC risk, it may enhance the effect of pri-miR-34b/c rs4938723 in women. rs4938723 CC genotype and rs11614913 TC genotype might predispose the host to immune selection of T1674C/G, and G1896A, respectively. The rs4938723 effect on HCC risk can be seriously affected by the HBV mutations. In light of our results, well-designed prospective studies with ethnically diverse HBV-infected populations and functional studies are warranted to elucidate the interaction of the SNPs with the HBV mutations in hepatocarcinogenesis.

Supporting Information Figure S1 Schematic diagram of altered binding sites of transcription factors in the putative promoter region of pri-miR-34b/c due to the T-to-C change at rs4938723. (TIF) Click here for additional data file.(78K, tif) Table S1 The primers, probes, and PCR program for genotyping the polymorphisms. (DOC) Click here for additional data file.(37K, doc) Table S2 Age, gender, and HBV infection-related parameters of the subjects enrolled in this study. (DOC) Click here for additional data file.(48K, doc) Table S3 The associations of the polymorphisms with HCC-free chronic HBV infection, LC, abnormal ALT and high viral load. (DOC) Click here for additional data file.(73K, Brefeldin_A doc) Table S4 Association of multiplicative interaction of pri-miR-34b/c rs4938723 and pre-miR-196a2 rs11614913 with HCC risk in multivariate regression analyses. (DOC) Click here for additional data file.(44K, doc) Acknowledgments Authors thank Drs. Rong Zhang, Jiaqi Zhang, Wu Ni, Xinyan Sun, Chengzhong Li, Qian Zhang, Huafen Wang, Lei Han for their help in the recruitment of the study subjects.