Kitchener H C, Neilson L, Macnab J C
Aberdeen Maternity Hospital, Foresterhill, U.K.
Eur J Cancer. 1993;29A(2):252-5. doi: 10.1016/0959-8049(93)90186-j.
Normal and abnormal biopsies of the uterine cervix, to a total of 124 samples, have been analysed for the detection of oestrogen receptor (ER) mRNA. The tough fibrous nature of the cervix proved very resistant to disaggregation in the first instance and subsequently, it was difficult to extract good high molecular weight message. This necessitated a systematic study of methodological technique, including two methods of tissue disaggregation and five techniques of extraction of ER mRNA, which in total involved the use of 124 cervical biopsies. The most successful method involved chopping the tissue, then digesting the cells with proteinase K and extracting the nucleic acids in salt and sodium dodecyl sulphate. Using the perfected technique, 16 cervical biopsies obtained at serial intervals from four women did not show any differences in ER mRNA in cervical biopsies either in the presence of oral contraception or histological abnormality. The successful method described will prove valuable for the detection of ER message in human tumours and other tissues of similar nature.
对总共124份子宫颈正常和异常活检样本进行了雌激素受体(ER)mRNA检测分析。子宫颈坚韧的纤维性质最初被证明很难分散,随后,很难提取到高质量的高分子量信息。这就需要对方法技术进行系统研究,包括两种组织分散方法和五种ER mRNA提取技术,总共涉及124份宫颈活检样本。最成功的方法是将组织切碎,然后用蛋白酶K消化细胞,并在盐和十二烷基硫酸钠中提取核酸。使用完善的技术,从四名女性身上按连续间隔获取的16份宫颈活检样本,在口服避孕药或组织学异常的情况下,宫颈活检样本中的ER mRNA均未显示出任何差异。所描述的成功方法对于检测人类肿瘤及其他类似性质组织中的ER信息将具有重要价值。