Oestrogen receptor message in premalignant and normal cervical cells: a methodological study.
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.
Kitchener
H C
HC
Aberdeen Maternity Hospital, Foresterhill, U.K.
Neilson
L
L
Macnab
J C
JC
D002584:Cervix Uteri / Q000737:chemistry; D019286:Endopeptidase K; D005260:Female; D006801:Humans; D011230:Precancerous Conditions / Q000737:chemistry*; D012333:RNA, Messenger / Q000032:analysis*; D011960:Receptors, Estrogen / Q000032:analysis*; D012697:Serine Endopeptidases; D002583:Uterine Cervical Neoplasms / Q000737:chemistry*
D012333:RNA, Messenger; D011960:Receptors, Estrogen; D012697:Serine Endopeptidases; D019286:Endopeptidase K