【摘要】 目的通过分析山东省青岛地区HBV核心启动子基因序列的突变特征,探讨其与乙型肝炎相关原发性肝癌的相关性。方法收取慢性乙型肝炎患者和乙型肝炎相关原发性肝癌患者的血清标本各60例,然后从中提取HBV DNA,采用聚合酶链反应(PCR)扩增、纯化、克隆后测序,根据S基因区的编码序列确定患者的基因型和血清型;分别将HBV核心启动子区的各序列结果与GeneBank中的HBV标准株做对比,用DNAMAN软件对基因序列进行突变分析。采用SPSS17.0软件进行统计学分析。结果 120例标本,HBV株均为B或C基因型,以C基因型为主,其中,HBV组的C基因型所占比率为83.33%,HCC组为90.00%(χ2=0.65,P=0.42);血清型均为adw2或adrq+。HBV基因组核心启动子区常见的点突变为C1653T、T1753V、C1754T及A1762T/G1764A,发生率分别为42.73%、86.36%、71.82%、38.18%。与慢性乙型肝炎患者相比,肝癌患者中发生率比较高的突变位点为C1653T(78.85%,χ2=52.58,P<0.001)及A1762T/G1764A(73.08%,χ2=50.88,P<0.001)。结论山东青岛地区HBV基因组常见为B、C基因型,其中以C型为主;核心启动子区突变发生率高,其中,T1753V与HCC发生无关,C1653T、A1762T和G1764A位点无论单独或是联合突变均与HCC发生密切相关。更多还原

【Abstract】 Objective To explore the relationship between HBV basal core promoter(BCP) mutations and primary liver cancer by analyzing the mutating features of HBV BCP in Qingdao,Shandong Province.Methods Serum specimens of patients with chronic hepatitis B(n=60) and hepatitis B related hepatocellular carcinoma(n=60) were collected.From each serum sample,HBV DNA was extracted.HBV S gene and BCP gene were amplified by polymerase chain reaction(PCR) and were cloned.HBV genotype and serotype were identified base on S gene sequence.The mutations were analysed with DNAMAN software by comparing the results with the standard strains of HBV in the GeneBank.SPSS17.0 software was used for statistical analysis.Rusults Among the samples(n=120),HBV strains could be categorized into genotype B or C,of which the major genotype was C-type,and serotypes was mainly adw2 and adrq +.The incidence of genotype C in patients with chronic hepatitis B and hepatitis B related hepatocellular carcinoma were 83.33% and 90.00%,respectively(χ2=0.65,P=0.42).The point mutations of T1753V(42.73%),C1653T(86.36%),C1754T(71.82%) and A1762T/G1764A(38.18%) were the major mutations in the HBV BCP.Compared with patients with chronic hepatitis B,point mutations C1653T(78.85%,χ2=52.58,P<0.001) and A1762T/G1764A(73.08%,χ2=50.88,P<0.001)in patients with liver cancer had a higher incidence.Conclusion HBV genotype B and C are the most prevalent genotypes,of which the major genotype is C-type;There is a high incidence of mutationin BCP gene,such as T1753V,may have no correlation with liver cancer and C1653T,A1762T,G1764A alleles alone or double mutantions might have a relationship with HCC in Qingdao,Shandong Province.更多还原