Shirakawa H
First Department of Surgery, Hokkaida University, School of Medicine, Sapporo, Japan.
Hokkaido Igaku Zasshi. 1996 Nov;71(6):677-88.
Chronic hepatitis C virus (HCV) infection has been clearly established as a major risk factor in the development of hepatocellular carcinoma. In the present study we have attempted to identify the HCV genotypes associated with hepatocellular carcinoma in patients from the USA and Japan. RNAs from tumorous and non-tumorous tissues from 11 HCV seropositive Japanese patients, and plasma from 4 American patients were analysed by reverse transcription-polymerase chain reaction (RT-PCR) methods employing primers specific for the 5'UTR, the NS5 and E2/NS1 regions. Amplified products were cloned and compared by nucleotide sequencing and phylogenetic analysis. The 5'UTR region could be successfully amplified and sequenced from all samples, and phylogenetic analysis of the nucleotide sequences demonstrated with the exception of two of Japanese viruses were closely related to HCV type 1. Type 2 was detected in these two cases. In addition, two of the Japanese patients who were found to have cholangiocarcinoma were also found to be infected with type 1. HCV amplification of the NS5 was successful in 7 of the Japanese and 1 USA sample and clearly demonstrated that genotype 1b was predominant. Amplification of the E2/NS1 regions proved to be extremely difficult and was unsuccessful in all HCC patients despite the fact that these regions could be consistently amplified in samples from patients with both acute and chronic HCV infection. These findings might suggest that with long term persistent HCV infection, there may be marked heterogeneity in both the structural and non-structural regions of the virus, and/or possibly that the viral genomes may be defective.
慢性丙型肝炎病毒(HCV)感染已被明确认定为肝细胞癌发生的主要危险因素。在本研究中,我们试图确定美国和日本患者中与肝细胞癌相关的HCV基因型。采用针对5'UTR、NS5和E2/NS1区域的特异性引物,通过逆转录-聚合酶链反应(RT-PCR)方法对11例HCV血清阳性日本患者的肿瘤组织和非肿瘤组织的RNA以及4例美国患者的血浆进行分析。将扩增产物克隆,并通过核苷酸测序和系统发育分析进行比较。所有样本的5'UTR区域均可成功扩增并测序,核苷酸序列的系统发育分析表明,除两株日本病毒外,其余均与1型HCV密切相关。在这两例中检测到2型。此外,还发现两名患有胆管癌的日本患者也感染了1型。NS5在7例日本样本和1例美国样本中成功扩增,明确显示1b型为主。尽管在急性和慢性HCV感染患者的样本中可以持续扩增E2/NS1区域,但在所有肝癌患者中,E2/NS1区域的扩增极其困难且未成功。这些发现可能表明,长期持续的HCV感染可能导致病毒的结构和非结构区域存在明显的异质性,和/或病毒基因组可能存在缺陷。