Hawkins R A, Hill A, Freedman B
Clin Chim Acta. 1975 Oct 15;64(2):203-10. doi: 10.1016/0009-8981(75)90202-8.
A simple method for the determination of oestrogen receptor activity in breast tumours and other tissues is described. By competition between radioactive and non-radioactive oestradiol-17beta for binding to a tissue extract prepared with centrifugation at low speed, the presence or absence of receptors is decided from the unmanipulated data. The concentration of receptors (P0) and dissociation constant of binding (Kd) are calculated by Scatchard analysis. In two relatively homogeneous tissues, assay precision was of the order of 16% for receptor concentration and 21-33% for the dissociation constant of binding: in heterogeneous breast tumors, much poorer precision can be expected. Sensitivity determined empirically is approximately 0.1 fmol/mg tissue. The method has been applied to the determination of receptor concentrations in human breast cancers, benign breast tumours, non-malignant breast, gynaecomastic breast, and in various tissues of the rat. Receptor activity was detected in 70% of the human breast cancers examined, and in lower amounts, in 21% of the benign human breast tissues. Activity was also detected in rat uterus and mammary gland, and in low amounts in 82% of the rat mammary tumours analysed. Dissociation constant of binding was generally of the order of 0.5 x 10(-10) M. The advantages of the method are discussed.
本文描述了一种测定乳腺肿瘤及其他组织中雌激素受体活性的简单方法。通过放射性和非放射性17β-雌二醇竞争结合经低速离心制备的组织提取物,根据未处理的数据判断受体的有无。受体浓度(P0)和解离常数(Kd)通过Scatchard分析计算得出。在两种相对均质的组织中,受体浓度的测定精度约为16%,结合解离常数的测定精度为21%-33%:在异质性乳腺肿瘤中,预期精度会差得多。根据经验确定的灵敏度约为0.1 fmol/mg组织。该方法已应用于测定人类乳腺癌、良性乳腺肿瘤、非恶性乳腺、男性乳腺增生症乳腺以及大鼠各种组织中的受体浓度。在所检测的人类乳腺癌中,70%检测到受体活性,在21%的良性人类乳腺组织中活性较低。在大鼠子宫和乳腺中也检测到活性,在82%分析的大鼠乳腺肿瘤中活性较低。结合解离常数一般约为0.5×10⁻¹⁰ M。文中讨论了该方法的优点。