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. 1982 Apr;35(4):401–406. doi: 10.1136/jcp.35.4.401

Biochemical assessment of histochemical methods for oestrogen receptor localisation.

J C Underwood, E Sher, M Reed, J A Eisman, T J Martin
PMCID: PMC497669  PMID: 7042761

Abstract

The biochemical validity of published labelled-ligand and ligand-antibody methods for the histological localisation of oestrogen receptors (ER) in breast carcinomas was assessed by measuring specific and non-specific binding of 3H-oestradiol to pretreated MCF-7 monolayers; the monolayers had been either frozen and thawed, then air-dried or acetone-fixed, or fixed in formaldehyde to reproduce the conditions in published methods. The results indicate that most of the ER in a frozen section is likely to be lost by diffusion into the supernatant incubation medium. Fixation substantially restricts diffusion but reduces specific oestradiol binding and may increase non-specific binding; acetone fixation is particularly deleterious. Preincubation in oestradiol fails to protect the receptor during fixation. We conclude that the published histochemical methods for ER localisation have doubtful validity and should not yet be used in preference to conventional biochemical assays.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barrows G. H., Stroupe S. B., Riehm J. D. Nuclear uptake of a 17 beta-estradiol-fluorescein derivative as a marker of estrogen dependence. Am J Clin Pathol. 1980 Mar;73(3):330–339. doi: 10.1093/ajcp/73.3.330. [DOI] [PubMed] [Google Scholar]
  2. Castaneda E., Liao S. The use of anti-steroid antibodies in the characterization of steroid receptors. J Biol Chem. 1975 Feb 10;250(3):883–888. [PubMed] [Google Scholar]
  3. Chamness G. C., Mercer W. D., McGuire W. L. Are histochemical methods for estrogen receptor valid? J Histochem Cytochem. 1980 Aug;28(8):792–798. doi: 10.1177/28.8.7003005. [DOI] [PubMed] [Google Scholar]
  4. Fishman J., Fishman J. H., Nisselbaum J. S., Menendez-Botet C., Schwartz M. K., Martucci C., Hellman L. Measurement of the estradiol receptor in human breast tissue by the immobilized antibody method. J Clin Endocrinol Metab. 1975 Apr;40(4):724–727. doi: 10.1210/jcem-40-4-724. [DOI] [PubMed] [Google Scholar]
  5. Ghosh L., Ghosh B. C., Das Gupta T. K. Immunocytological localization of estrogen in human mammary carcinoma cells by horseradish--anti-horseradish peroxidase complex. J Surg Oncol. 1978;10(3):221–224. doi: 10.1002/jso.2930100308. [DOI] [PubMed] [Google Scholar]
  6. Greene G. L., Nolan C., Engler J. P., Jensen E. V. Monoclonal antibodies to human estrogen receptor. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5115–5119. doi: 10.1073/pnas.77.9.5115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horwitz K. B., Costlow M. E., McGuire W. L. MCF-7; a human breast cancer cell line with estrogen, androgen, progesterone, and glucocorticoid receptors. Steroids. 1975 Dec;26(6):785–795. doi: 10.1016/0039-128x(75)90110-5. [DOI] [PubMed] [Google Scholar]
  8. Kurzon R. M., Sternberger L. A. Estrogen receptor immunocytochemistry. J Histochem Cytochem. 1978 Oct;26(10):803–808. doi: 10.1177/26.10.363929. [DOI] [PubMed] [Google Scholar]
  9. Lee S. H. Cytochemical study of estrogen receptor in human mammary cancer. Am J Clin Pathol. 1978 Aug;70(2):197–203. doi: 10.1093/ajcp/70.2.197. [DOI] [PubMed] [Google Scholar]
  10. Lippman M. E., Bolan G. Oestrogen-responsive human breast cancer in long term tissue culture. Nature. 1975 Aug 14;256(5518):592–593. doi: 10.1038/256592a0. [DOI] [PubMed] [Google Scholar]
  11. Mercer W. D., Lippman M. E., Wahl T. M., Carlson C. A., Wahl D. A., Lezotte D., Teague P. O. The use of immunocytochemical techniques for the detection of steroid hormones in breast cancer cells. Cancer. 1980 Dec 15;46(12 Suppl):2859–2868. doi: 10.1002/1097-0142(19801215)46:12+<2859::aid-cncr2820461425>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  12. Morrow B., Leav I., Delellis R. A., Raam S. Use of polyestradiol phosphate and anti-17 beta estradiol antibodies for the localization of estrogen receptors in target tissues: a critique. Cancer. 1980 Dec 15;46(12 Suppl):2872–2879. doi: 10.1002/1097-0142(19801215)46:12+<2872::aid-cncr2820461427>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  13. Nenci I., Beccati M. D., Piffanelli A., Lanza G. Detection and dynamic localisation of estradiol-receptor complexes in intact target cells by immunofluorescence technique. J Steroid Biochem. 1976 Jun-Jul;7(6-7):505–510. doi: 10.1016/0022-4731(76)90194-1. [DOI] [PubMed] [Google Scholar]
  14. Pertschuk L. P. Detection of estrogen binding in human mammary carcinoma by immunofluorescence: a new technic utilizing the binding hormone in a polymerized state. Res Commun Chem Pathol Pharmacol. 1976 Aug;14(4):771–774. [PubMed] [Google Scholar]
  15. Pertschuk L. P., Gaetjens E., Carter A. C., Brigati D. J., Kim D. S., Fealey T. E. An improved histochemical method for detection of estrogen receptors in mammary cancer. Comparison with biochemical assay. Am J Clin Pathol. 1979 May;71(5):504–508. doi: 10.1093/ajcp/71.5.504. [DOI] [PubMed] [Google Scholar]
  16. Pertschuk L. P., Tobin E. H., Brigati D. J., Kim D. S., Bloom N. D., Gaetjens E., Berman P. J., Carter A. C., Degenshein G. A. Immunofluorescent detection of estrogen receptors in breast cancer. Comparison with dextran-coated charcoal and sucrose gradient assays. Cancer. 1978 Mar;41(3):907–911. doi: 10.1002/1097-0142(197803)41:3<907::aid-cncr2820410318>3.0.co;2-v. [DOI] [PubMed] [Google Scholar]
  17. Rao B. R., Fry C. G., Hunt S., Kuhnel R., Dandliker W. B. A fluorescent probe for rapid detection of estrogen receptors. Cancer. 1980 Dec 15;46(12 Suppl):2902–2906. doi: 10.1002/1097-0142(19801215)46:12+<2902::aid-cncr2820461432>3.0.co;2-o. [DOI] [PubMed] [Google Scholar]
  18. Taylor C. R., Cooper C. L., Kurman R. J., Goebelsmann U., Markland F. S., Jr Detection of estrogen receptor in breast and endometrial carcinoma by the immunoperoxidase technique. Cancer. 1981 Jun 1;47(11):2634–2640. doi: 10.1002/1097-0142(19810601)47:11<2634::aid-cncr2820471119>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  19. Walker R. A., Cove D. H., Howell A. Histological detection of oestrogen receptor in human breast carcinomas. Lancet. 1980 Jan 26;1(8161):171–173. doi: 10.1016/s0140-6736(80)90660-1. [DOI] [PubMed] [Google Scholar]

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