原发性肝癌患者乙型肝炎病毒基因型与突变的地区性差异分析

目的 研究原发性肝癌患者乙型肝炎病毒(HBV)基因型、亚型和突变的地区性差异.方法 HBV阳性的原发性肝癌组织标本80例,其中广西地区20例,上海、浙江、江苏(长三角地区)60例.应用聚合酶链反应(PCR)扩增患者携带的HBV DNA序列,PCR产物直接纯化测序.用NCBI在线的软件viral genotyping tool确定HBV基因型,MEGA(Molecular Evolutionary Genetics Analysis,version 3.1)软件构建系统进化树和进行亚型分型及突变分析.结果 80例肝癌标本中,17例为B基因型(21.3%),57例为C基因型(71.3%),6例为B、C基因型混合感染(7.5%).对17例B基因型和57例C基因型的亚型分型,其中B基因型的亚型全部为B2型,C基因型的亚型有C1、C2、C5等三型,其中有12例为C1型(21.1%),44例为C2型(77.2%),1例为C5型(1.8%).广西地区的C1亚型35%(7/20)明显高于长三角地区的C1亚型8.5%(5/60),而广西地区的C2亚型20%(4120)明显低于长三角地区的C2亚型66.7%(40160).两地区HBV基因亚型分布在统计学上有显著差异.比较两地区的HBV序列还发现,广西地区B基因型CP区T1762/A1764呈现更高的突变性,有6例(100%)存在突变,而长三角地区只有2例(16.7%).结论 两地区原发性肝癌患者的HBV亚型分布和基因型B的CP区T1762/A1764突变率有显著差异.

Genotyping and mutation analysis of hepatitis B virus in hepatocellular carcinoma patients from different areas in China

Objective To study hepatitis B virus (HBV) genotype and mutation in hepatocellular carcinoma (HCC) cases in Guangxi area and Changsanjiao area.Methods we used PCR to detect HBV genotype of 80 HBV-infected HCC cases from two areas including 20 cases from Guangxi area and 60 cases from Changsanjiao area (Shanghai, Zhejiang, Jiangsu). Data were analyzed by NCBI online software viral genotyping tool and phylogenetic tree of HBV sequences constructed by MEGA (version 3.1) software.Results Among 80 HBV-infected HCC cases, the prevalence of genotype B,C, and B/C were 21.3%(17/80),71.3% (57/80) and 7.5% (6/80), respectively. In genotype C, 12 cases were subtype C1(21.1%), 44 cases were subtype C2 (77.2%), 1 case was subtype C5 (1.8%). In genotype B, all cases were B2. In Guangxi area, the prevalence of subtype C1,C2 were 35% (7/20) and 20% (4/20). In Changsanjiao area, the prevalence of subtype C1,C2 were 8.5% (5/60) and 66.7% (40/60). The HBV subtype C1, C2 distribution of two area exhibited significant differences in the HBV-infected HCC cases. In addition, the T1 762/A1 764 mutation of two areas exhibited significant differences in genotype B, in Guangxi area, the prevalence of mutation in genotype B was 100% (6/6), and in Changsanjiao area, the prevalence was 16.7% (2/11).Conclusion HBV subtypes B2, C1, C2, C5 distribution and BCP mutation in genotype B showed significant differences between HBV-infected HCC cases of two areas.