Study of HBV- X gene mutation among patients with HBV- related chronic hepatitis, liver cirrhosis, and primary liver cancer
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摘要: 目的研究慢性HBV感染者,如慢性乙型肝炎(CHB)、乙型肝炎肝硬化(LC)、原发性肝癌(PLC)患者血清中HBV-X基因序列的突变与肝癌发生的关系。方法收集2011-2013年间于重庆医科大学附属第二医院就诊的慢性HBV感染者血清共89例,从血清中提取HBV DNA,扩增全长HBV-X基因序列,经测序后与已知HBV-X基因相应序列比较该患者体内HBV-X基因变异位点以及变异形式,并用卡方检验、单因素方差分析处理数据,NCBI的genotype工具测定基因型。结果所有患者均属于B/C基因型,HBeAg阳性患者中B基因型占46.2%,C基因型占53.8%;HBeAg阴性患性中B基因型占81.2%,C基因型占18.8%(P=0.001)。在PLC组中,启动子(BCP)区的突变显著高于CHB、LC(69.2%vs34.4%和61.3%,P<0.05),且nt1821位点存在明显的T碱基的缺失(88.5%vs 53.1%和71%,P=0.014)。在CHB、LC中,C基因型BCP的双突变率显著高于B基因型(61.5%vs 15.8%,P=0.007;83.3%vs 47.4%,P=0.04...Abstract: Objective To study the relationship between hepatocarcinogenesis and the mutation in X gene among patients with chronic hepatitis B virus ( HBV) infection, such as chronic hepatitis B ( CHB) , liver cirrhosis ( LC) and primary liver cancer ( PLC) . Methods The serum samples from 89 patients with chronic HBV infection who visited the Second Affiliated Hospital of Chongqing Medical University from2011 to 2013 were collected. PCR was used to amplify the X gene of HBV DNA extracted from the serum samples. After sequencing, the HBV- X genome was compared with those reported in GenBank to find the variable sites and variant forms. Chi- square and one- way ANOVA were used for the statistical analysis afterwards, whereas genotypes were determined by the genotyping tool of the National Center for Biotechnology Information. Results All patients were genotype B or C. Among HBeAg- positive patients, 46. 2% were genotype B, and53. 8% were genotype C; among HBeAg- negative patients, 81. 2% were genotype B, and 18. 8% were genotype C ( P = 0. 001) . PLC patients had a significantly higher risk of mutation in the basic core promoter ( BCP) region than the CHB and LC groups ( 69. 2% vs 34. 4%and 61. 3%, P < 0. 05) ; in addition, an evident T- base deficiency was observed at nt1821 site ( 88. 5% vs 53. 1% and 71%, P =0. 014) . Among CHB and LC patients, those with genotype C had a significantly higher risk of BCP double mutation than those with genotype B ( 61. 5% vs 15. 8%, P = 0. 007; 83. 3% vs 47. 4%, P = 0. 045) . The incidence of BCP double mutation was significantly higher in the low- viral load group ( ≤106copies /ml) than in the high- viral load group ( > 106copies /ml) ( 81. 3% vs 47. 9%, P = 0. 015) .Conclusion The BCP double mutation and T- base deficiency at nt1821 site may play important roles in the development of PLC.
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Key words:
- hepatitis B, chronic /
- liver cirrhosis /
- carcinoma, hepatocellular /
- hepatitis B virus /
- point mutation
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