论文部分内容阅读
目的:为探讨丙型肝炎病毒(HCV)感染在肝癌发生中的作用。方法:从HCV第二代抗体阳性的肝癌和慢性肝炎患者血清中提取HCV-RNA,经随机引物逆转录合成cDNA,再以病毒5’-末端至E1区间的多对引物进行巢式PCR扩增,其最终产物(425bp和621bp)分别连接p-GEM-T载体,在大肠杆菌中表达后进行测序。结果:所有患者均为HCV-Ⅱ型感染,完整的核心蛋白基因(573bp)与已报道的其它株比较,其核苷酸同源性为:慢性肝炎株89.0%~95.5%,肝癌株88.7%~92.3%;氨基酸同源性为:前者91.1%~96.6%,后者87.4%~92.1%。且发现该段基因中核苷酸置换肝癌患者较明显地高于慢性肝炎患者。结论:本文资料提示核心蛋白基因的变异,可能与HCV的慢性持续感染及肝癌的发生有关。
Objective: To investigate the role of hepatitis C virus (HCV) infection in the development of hepatocellular carcinoma. METHODS: HCV-RNA was extracted from the serum of HCV second-antibody positive liver cancer patients and chronic hepatitis patients. cDNA was reverse transcribed by random primers and nested PCR amplification was performed using multiple pairs of primers from the 5′-end to the E1 interval of the virus. The final products (425 bp and 621 bp) were respectively ligated into the p-GEM-T vector and expressed in E. coli and then sequenced. Results: All patients were infected with HCV-II, and the complete core protein gene (573 bp) was compared with other reported strains. The nucleotide homology was: chronic hepatitis strains 89.0%-95.5%, liver cancer strains 88.7% to 92.3%; the amino acid homology was 91.1% to 96.6% in the former and 87.4% to 92.1% in the latter. It was found that the patients with nucleotide replacement liver cancer in this segment of the gene were significantly higher than those with chronic hepatitis. Conclusion: The data in this paper suggest that the variation of the core protein gene may be related to the chronic persistent infection of HCV and the occurrence of liver cancer.