Menegazzi M, Scarpa A, Libonati M
Istituto di Chimica Biologica, Facoltà di Medicina e Chirurgia, Università di Verona.
Ital J Biochem. 1988 Mar-Apr;37(2):104-10.
To determine the methylation pattern of c-Ha-ras oncogene in prostatic tissue and to identify possible changes of methylation associated with cancer, high molecular weight DNA was extracted from 7 normal and 6 carcinomatous human prostates. Analysis of the samples was performed by cleaving DNA with the restriction endonucleases Msp I, Hpa II and Cfo I, and by Southern hybridizing the DNA digests with the 32P-labelled c-Ha-ras (pT24-C3) probe. Several discrete fragments were obtained with Hpa II and Cfo I digestion while the Msp I pattern showed fewer and smaller bands. Therefore, c-Ha-ras appears to be partially methylated. While a considerable polymorphism of the sequence 5'-CCGG-3' was observed at several Msp I sites in all cases, no significant differences could be evidenced in the methylation patterns of normal and neoplastic prostatic DNA samples extracted and purified from each patient.
为了确定前列腺组织中c-Ha-ras癌基因的甲基化模式,并识别与癌症相关的甲基化可能发生的变化,从7例正常人类前列腺和6例癌性人类前列腺中提取了高分子量DNA。通过用限制性内切酶Msp I、Hpa II和Cfo I切割DNA,并将DNA消化产物与32P标记的c-Ha-ras(pT24-C3)探针进行Southern杂交来对样本进行分析。用Hpa II和Cfo I消化得到了几个离散的片段,而Msp I模式显示的条带更少且更小。因此,c-Ha-ras似乎部分甲基化。虽然在所有病例的几个Msp I位点均观察到5'-CCGG-3'序列存在相当大的多态性,但从每位患者提取和纯化的正常和肿瘤性前列腺DNA样本的甲基化模式中未发现显著差异。