慢性丙型肝炎患者HCV核心蛋白基因及其肽链的序列分析  

SEQUENCE ANALYSES OF CORE GENES OF HEPATITIS C VIRUS GENOME AND THEIR PEPTIDE CHAINS IN PATIENTS WITH CHRONIC HEPATITIS C

作　　者：姚登福[1,2] 孟宪镛[1,2] 黄介飞[1,2] 蒋道荣 倪润洲[1,2] 清水一郎 伊东进[1,2] 

机构地区：[1]南通医学院附属医院临床分子生物学中心 [2]日本国德岛大学第二内科

出　　处：《南通医学院学报》1998年第2期135-138,共4页ACTA Academiae Medicinae Nantong

基　　金：日本国文部省资助

摘　　要：为探讨丙型肝炎病毒（ＨＣＶ）感染在肝癌发生中的作用，从３５例ＨＣＶ－ＲＮＡ阳性的非甲非乙型肝病患者血清中提取ＨＣＶ－ＲＮＡ，经随机引物逆转录合成ｃＤＮＡ，在病毒的Ｃ基因区进行分型，再于病毒５＇－末端至Ｅ１区间以多对引物进行巢式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分析。ＨＣＶ基因型表现为８２％Ⅱ型，９％Ⅲ型及９％Ⅱ＋Ⅲ混合型。核心蛋白基因（５７３ｂｐ）与已报道的其他株比较，其核苷酸同源性为：慢性肝炎株９３．０±２．４％，肝癌株９１．４±１．５％；氨基酸同源性为：前者９３．９±０．８％，后者９１．０±１．６％；核心蛋白基因中核苷酸置换在肝癌株较明显地高于慢性肝炎株。研究结果提示，ＨＣＶ核心蛋白基因的错义突变，与ＨＣＶ的慢性持续感染及肝癌发生的关系密切。To explore the role of hepatitis C virus (HCV) in the causation of hepatocellular carcinoma (HCC), HCV-RNAs extracted from the sera of 35 patients with positive for HCV-RNA were converted to cDNA by reverse transcription with random primer and genotyped in the core gene region of HCV genome, PCR assay was performed with the primers between 5'-NC and E1 region, Amplified products of 425 bp and 621 bp were expressed in E. coli with p-GEM-T plasmis vector, and their nucleotide sequences were deterimined by dideoxynucleotide chain-termination method. GenotypeⅡ, Ⅲ and Ⅱ/Ⅲ was 82%, 9% and 9%, respectively. The comparison of the core gene sequences (573bp) with other isolates reported revealed that the homology is 93.0±2.4% in CH, 93.9±0.8% in HCC for nucleotide sequences and 91.4±1.5% in CH, 91.0±1.6% in HCC for amino acid sequences, respectively. The results showed a tendency for more nucleotide substitions in the core gene in HCC patients than in those with CH. The present suggests that missense mutations in the core gene region are closely related with HCV chronic persistent infection and progression of chronic liver disease to HCC.

关 键 词：丙型肝炎病毒 C基因 序列分析 基因表达 肽链 

分 类 号：R373.21[医药卫生—病原生物学] R512.630.2[医药卫生—基础医学]