肝癌病人血清中丙型肝炎病毒E2／NS1基因区cDNA的克隆及序列分析

CLONING AND SEQUENCING OF cDNA OF E2/NS1 GENE OFHEPATITIS C VIRUS FROM A PATIENT WITHHEPATOCELLULAR CARCINOMA

从一个抗丙型肝炎病毒（ＨＣＶ）阳性的肝细胞癌（ＨＣＣ）病人血清中提取ＲＮＡ，随机引物逆转录为ｃＤＮＡ后，用ＨＣＶ特异引物进行聚合酶链反应。将扩增产物７８０ｂｐ插入ｐＵＣ１８和ｐＵＣ１９质粒载体，双脱氧链末端终止法测定其序列，与慢性丙型肝炎病人或携带者血清中的ＨＣＶ序列比较，核苷酸同源性介乎６９．２３％－８９．１０％，氨基酸同源性介乎７４，５９％－９０．５７％、分析表明，此序列属于１组Ⅱ型。本文结果有助于了解ＨＣＶ在ＨＣＣ发生中的作用。

epatitis C virus (HCV) RNA extracted from the serum of a patient with hepatocellularcarcinoma (HCC)which contained anti- HCV was converted to cDNA by reverse transcriptionwith random primer.Polymerase chain reaction was performed with the primers which spanthe HCV E2/NS1 region.Amplified products of 780bp was ligated to pUC18 and pUC19plasmid vector,and their nucleotide sequences detemined by dideoxy-mediated chaintermination method.A comparison of the sequence with several previously reported isolatesderived from patients with chromic non-A non-B hepatitis or asymptomatic carriers showsthat the homology is 69.23%- 89.10% for nucleotide sequence and 74.59%- 90.57% foramino acid sequence.The isolate we obtained was identified as genotype Ⅱof group 1 .Thisstudy may be helpful in clarifying the role of HCV in the causation of HCC.