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Standardization of Steroid Receptor Analysis in Breast Cancer Biopsies: EORTC Receptor Group

  • A. Koenders
  • T. J. Benraad
Part of the Recent Results in Cancer Research book series (RECENTCANCER, volume 91)

Abstract

In 1972 several members of the EORTC Breast Cancer Cooperative Group agreed on the potential value of estrogen receptor (ER) determinations in human mammary carcinoma for therapeutic guidance [3]. During that workshop standards for the assessment of these assays were proposed. The charcoal adsorption technique was recommended as a first standard procedure, and the receptor concentration was to be calculated according to Scatchard. In 1978 the EORTC Breast Cancer Cooperative Group decided that the ER content of the tumor would determine the entry of breast cancer patients into some of the future clinical trials. To achieve adequate and uniform methodology of the ER assay, a workshop was held in 1979 [4]. The purpose of this workshop was to discuss practical aspects of techniques of measuring ER and moreover to decide upon a single standardized receptor assay. Some of the most important steps agreed upon with regard to the receptor binding assay can be summarized as follows:
  1. 1.

    The composition of the extraction buffer (10 mM HPO4/H2PO4, 1.5 mM EDTA, 3 mM NaN3, 10 mM monothioglycerol, 10% v/v glycerol; pH7.5).

     
  2. 2.

    High-speed centrifugation, resulting in particle-free extracts, is preferred over low-speed centrifugation.

     
  3. 3.

    The cytosol protein concentration should not be less than 1 mg/ml.

     
  4. 4.

    The minimal purity of the radioactive ligands should be 95%.

     
  5. 5.

    The recommended ligands for the ER determination are estradiol and moxestrol, while R-5020 or Org-2058 should preferentially be used for the progesterone receptor (PgR) assay.

     
  6. 6.

    The nonspecific binding is to be determined in the presence of a 100-fold excess of unlabeled diethylstilbestrol for the ER assay and R-5020 or 0rg-2058 for the PgR assay.

     
  7. 7.

    The charcoal adsorption technique combined with Scatchard analysis was to be retained as the standard procedure.

     
  8. 8.

    The receptor concentration is expressed as fmol/mg protein. Protein is measured according to Bradford [2], using Kabi human albumin as a protein standard. The Coomassie-blue reagent is obtained from Bio-Rad.

     

Keywords

Scatchard Analysis Uterine Tissue Receptor Assay Protein Result Tissue Powder 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Benraad T, Koenders A (1980) Estradiol receptor activity in lyophilized calf uterus and human breast tumor tissue. Cancer 46: 2762–2764PubMedCrossRefGoogle Scholar
  2. 2.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72: 248–254PubMedCrossRefGoogle Scholar
  3. 3.
    EORTC Breast Cancer Cooperative Group (1973) Standards for the assessment of estrogen receptors in human breast cancer. Eur J Cancer 9: 379–381CrossRefGoogle Scholar
  4. 4.
    EORTC Breast Cancer Cooperative Group (1980) Revision of the standards for the assessment of hormone receptors in human breast cancer. 2nd EORTC workshop, 16-17 March, 1979. Eur J Cancer 16: 1513–1515CrossRefGoogle Scholar
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    Koenders AJ, Geurts-Moespot J, Kho KH, Benraad TJ (1978) Estradiol and progesterone receptor activities in stored lyophilised target tissue. J Steroid Biochem 9: 947–950PubMedCrossRefGoogle Scholar
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    Raam S, Gelman R, Cohen JL (1981) Estrogen receptor assay: Interlaboratory and intralaboratory variations in the measurement of receptor using dextran-coated charcoal technique: a study sponsored by ECOG. Eur J Cancer 17: 643–649PubMedCrossRefGoogle Scholar
  7. 7.
    Wagner RK, Jungblut PW (1980) Quality control in steroid hormone receptor analyses. Cancer 46: 2950–2952PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin · Heidelberg 1984

Authors and Affiliations

  • A. Koenders
    • 1
  • T. J. Benraad
    • 1
  1. 1.Department of Experimental and Chemical Endocrinology, Medical FacultyUniversity of NijmegenNijmegenThe Netherlands

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