Polymorphism analyses of hepatitis B virus X gene in hepatocellular carcinoma patients from southern China

Chronic hepatitis B virus(HBV)infection is one of the major causes of hepatocellular carcinoma(HCC),and the HBV X(HBx)gene plays a critical role in the molecular pathogenesis of HBV-related HCC.We have investigated whether there are particular HBx gene mutations associated with HCC in patients from southern China.The HBx gene was examined in 51 paraffin-embedded tumor tissue samples from patients with HCC and 25 serum samples from the HBV carrier by nested polymerase chain reaction(PCR),single-stranded conformational polymorphism and heteroduplex analysis.The HBx genes with potentially important mutations from tumor tissue samples were cloned,sequenced and aligned with the published HBx gene sequence.HBV genotypes in tumor tissue samples were analyzed by nested PCR. Analyses of HBx gene polymorphism showed that 31.3% of HBx gene fragments in tumor tissue samples had a special pattern.A common deletion at nt 382-400 of the HBx gene accompanied by 29 point mutations was detected in four randomly selected tumor tissue samples with this pattern which caused a frame-shift in the HBx open reading frame with a new stop codon at nt 1818,resulting in an HBx polypeptide chain truncated at the C end in these cases.Among the four randomly selected samples,three were HBV genotype B,and one was not detected by our present assay.In another tumor tissue sample,amplification of the full-length HBx gene yielded a shorter fragment.Sequencing of this fragment revealed a 264 bp deletion between nt 1577 and 1840 of the HBV gene.These results suggest that HBx gene mutation occurs frequently in HCC samples,and the deletion at nt 382-400 of the HBx gene might play a role in carcinogenesis of HCC in southern China.