Leon M, Kew M C
Medical Research Council/Witwatersrand University, Department of Medicine, Johannesburg, South Africa.
Anticancer Res. 1995 May-Jun;15(3):859-61.
Paired samples of hepatocellular carcinoma and non-tumorous liver tissue from 12 southern African blacks were examined for mutations in codons 12, 13, and 61 of the three ras proto-oncogenes (H-, K-, and N-ras). Deoxyribonucleic acid was isolated from carcinoma and non-tumorous tissues and amplified with the polymerase chain reaction. Using the single-stranded conformational polymorphisms method, products of the polymerase chain reaction amplification of codons 12, 13, and 61 of H-, K-, and N-ras were analysed for mutations. Mobility shifts were not detected except in one tumour in the region of codon 61 of K-ras. By sequencing the relevant polymerase chain reaction products, this sequence of deoxyribonucleic acid was proved to be normal, indicating that the single-stranded conformational polymorphisms result was an artifact of the polymerase chain reaction. Thus, no mutations were detected in the regions of interest in any of the tumours studied. These results indicate that activation of ras proto-oncogenes by mutations in codons 12, 13, and 61 does not play an important role in hepatocellular carcinogenesis in southern African Blacks despite the fact that dietary exposure to aflatoxin B1 is a risk factor in this population.
对12名南非黑人的肝细胞癌组织和非肿瘤性肝组织配对样本进行检测,以查找三种原癌基因(H-ras、K-ras和N-ras)第12、13和61密码子的突变情况。从癌组织和非肿瘤组织中分离出脱氧核糖核酸,并采用聚合酶链反应进行扩增。利用单链构象多态性方法,对H-ras、K-ras和N-ras第12、13和61密码子的聚合酶链反应扩增产物进行突变分析。除了在一个肿瘤中检测到K-ras第61密码子区域存在迁移率改变外,未检测到其他迁移率改变。通过对相关聚合酶链反应产物进行测序,证实该脱氧核糖核酸序列正常,这表明单链构象多态性结果是聚合酶链反应的假象。因此,在所研究的任何肿瘤的感兴趣区域均未检测到突变。这些结果表明,尽管黄曲霉毒素B1的膳食暴露是该人群的一个危险因素,但第12、13和61密码子的突变激活ras原癌基因在南非黑人肝细胞癌发生过程中并不起重要作用。