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目的分析肝细胞癌组织中乙肝病毒HBx-d382突变体S基因和前S基因,了解其变异情况。方法采用巢式PCR和琼脂糖凝胶电泳的方法对16份肝细胞癌组织中乙肝病毒S基因和前S基因进行扩增和分析。选择HBxd382突变体S基因和前S基因目的片段进行回收、纯化、克隆和测序,并与HBV参考序列进行比对。结果16份肝癌组织中有9份存在HBx-d382突变体,HBx-d382突变体肝癌组织S基因和前S基因检出率分别为11.11%(1/9)、44.44%(4/9),且50.00%的肝癌组织前S基因PCR扩增产物出现了多条带。HBx-d382突变体S基因和前S基因目的片段与HBV参考序列(AF282917)比对,一致性介于98.08%至98.97%。结论 HBx-d382突变体肝癌组织中部分检出了S基因和前S基因,前S基因比S基因更容易发生突变。
Objective To analyze the S gene and pre-S gene of hepatitis B virus HBx-d382 mutant in hepatocellular carcinoma and to understand its variation. Methods The nested PCR and agarose gel electrophoresis were used to amplify and analyze hepatitis B virus S gene and pre-S gene in 16 hepatocellular carcinoma tissues. HBxd382 mutant S gene and the pre-S gene target fragment was recovered, purified, cloned and sequenced, and compared with the HBV reference sequence. Results HBx-d382 was found in 9 of 16 HCC tissues. The positive rate of S gene and pre-S gene in HBx-d382 mutant hepatocellular carcinoma was 11.11% (1/9) and 44.44% (4/9), respectively. And 50.00% of HCC pre-S gene PCR amplification products appeared in multiple bands. HBx-d382 mutant S gene and pre-S gene fragment and HBV reference sequence (AF282917) alignment, the consistency of 98.08% to 98.97%. Conclusion The S gene and pre-S gene were partially detected in the HBx-d382 mutant hepatocellular carcinoma. The pre-S gene was more likely to mutate than the S gene.