Vollmer G, Bindewald I, Meyn U, Wünsche W O, Arnholdt H, Knuppen R
Institut für Biochemische Endokrinologie, Medizinische Universität, Lübeck, F.R.G.
J Steroid Biochem. 1988 Dec;31(6):899-907. doi: 10.1016/0022-4731(88)90331-7.
An improved radioreceptor assay of unfixed cryostat sections of human target tissues has been developed. Sections collected on glass coverslips were immediately incubated with 5 nM concentrations of either tritiated estradiol-17 beta for estrogen receptor (ER) or ORG 2058 for progesterone receptor (PR) determination. For quantitation, receptor-bound and free hormone were separated by isoelectric focusing (IEF). The assay allows the determination of steroid hormone receptors and comparative histological examinations in immediately neighbouring serial sections of a single piece of tissue. Biochemically, the validity of the assay procedure was evidenced by Scatchard analysis, by ligand and tissue specificities, by the linear relations of receptor and protein concentrations and the number of sections per test tube. Diagnostically, we compared the routine (6 point DCC-Scatchard) procedure for breast cancer analysis with the section method. A good correlation for ER and a less pronounced correlation for PR was found. Statistically, the precision of the method was verified by low deviations of duplicate determinations, low day-to-day variations and low inter-assay variations.
已开发出一种用于人靶组织未固定低温切片的改进型放射受体测定法。收集在玻璃盖玻片上的切片立即与5 nM浓度的用于雌激素受体(ER)测定的氚化雌二醇-17β或用于孕激素受体(PR)测定的ORG 2058一起孵育。为了进行定量,通过等电聚焦(IEF)分离受体结合的激素和游离激素。该测定法允许在同一块组织的紧邻连续切片中测定类固醇激素受体并进行比较组织学检查。从生化角度来看,该测定程序的有效性通过Scatchard分析、配体和组织特异性、受体与蛋白质浓度的线性关系以及每个试管中的切片数量得到了证明。在诊断方面,我们将乳腺癌分析的常规(六点DCC-Scatchard)程序与切片法进行了比较。发现ER的相关性良好,而PR的相关性不太明显。从统计学角度来看,该方法的精密度通过重复测定的低偏差、低日间变化和低批间变化得到了验证。