肝癌组织中HBV核心基因启动子突变的研究

The Discovery of HBV Core Promoter Mutations in Tumour Tissue of Patients with Hepatocellular Carcinoma

为深入了解乙肝病毒 (HBV)致癌机理 ,用套式PCR对广西 14例血清HBVDNA阳性的原发性肝癌患者癌组织、10例乙型病毒性肝炎患者血清及 10例乙肝病毒无症状携带者血清HBV核心基因启动子进行扩增 ,阳性者用Sanger氏双脱氧法做序列分析 ,发现肝癌组织 10例PCR阳性 ,阳性率 71.4 % ,所有PCR阳性标本的整合体均有乙肝病毒核心基因启动子双突变序列 (nt 176 2A→T ,176 4G→A) ,并且在其周围各序列都有不同部位的点突变 ,标本C14核苷酸的缺失突变高达10个。乙肝患者 6例PCR阳性 ,其中 3例乙肝病毒核心基因启动子发生双突变。无症状携带者中 4例PCR阳性 ,其中仅 1例发生双突变。结果提示乙肝病毒核心基因启动子双突变在肝癌患者中较常见 ,肝炎患者次之 ,无症状携带者居最后。

In order to understand the mechanisms of hepatitis B virus (HBV) in the development of hepatocellular carcinoma, the nested PCR was used to amplify HBV core promoter from tumour tissues of 14 HBV DNA positive patients with hepatocellular carcinoma, and 10 serum samples of hepatitis B patients and asymptomatic carriers respectively. The positive samples were sequenced by Sanger method. Ten of the fourteen tumour tissue samples (71.4%) were positive for HBV core promoter. Each sequence of all positive samples had the double mutations at nt 1762(A→T), 1764(G→A) in HBV core promoter and some point mutations around them. The case C14 had more extensive mutations, including deletion of 10 nucleotides. 6 of 10 serum samples of hepatitis B patients were positive for HBV core promoter and 3 of them had the double mutations. four of the ten serum samples of asymptomatic carriers were PCR positive while only 1 of them had the double mutations. These data implicate that the double mutations is most common in HCC patients, the next is hepatitis B patients and the last is asymptomatic carriers.