Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1988 Jun;41(6):623–632. doi: 10.1136/jcp.41.6.623

Quantification of oestrogen receptors in breast cancer: radiochemical assay on cytosols and cryostat sections compared with semiquantitative immunocytochemical analysis.

H M Scheres 1, A F De Goeij 1, M J Rousch 1, G G Hondius 1, D D Willebrand 1, A H Gijzen 1, F T Bosman 1
PMCID: PMC1141542  PMID: 3384997

Abstract

A radiochemical oestrogen receptor assay on cytosol was correlated with a radiochemical and an immunohistochemical oestrogen receptor assay using cryostat sections from 50 breast cancer specimens. Oestrogen receptors were reliably quantitated in 6 micron cryostat sections with Scatchard analysis using radiolabelled oestradiol, and a good quantitative and qualitative relation with cytosol oestrogen receptor assay was found. Parallel sections were used for routine histological tissue verification and for direct comparison with immunohistochemistry for oestrogen receptor. Specific immunoperoxidase staining with a rat monoclonal antibody was scored by semiquantitative evaluation of the staining intensity of cancer cell nuclei. Oestrogen receptor scoring was highly reproducible when performed by the same observer. The semiquantitative immunohistochemical oestrogen receptor score correlated significantly better with the radiochemical assay on sections than with cytosol assay. Oestrogen receptor in breast cancer can be reliably assayed by semiquantitative evaluation of cryostat sections immunostained for oestrogen receptor, but only if the procedure is adequately standardised. The results underline the importance of cellular heterogeneity as a cause of variation in oestrogen receptor assay evaluation in breast cancer.

Full text

PDF
626

Selected References

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

  1. Allegra J. C., Lippman M. E., Thompson E. B., Simon R., Barlock A., Green L., Huff K. K., Do H. M., Aitken S. C., Warren R. Estrogen receptor status: an important variable in predicting response to endocrine therapy in metastatic breast cancer. Eur J Cancer. 1980 Mar;16(3):323–331. doi: 10.1016/0014-2964(80)90348-5. [DOI] [PubMed] [Google Scholar]
  2. Andersen J., Orntoft T., Poulsen H. S. Semiquantitative oestrogen receptor assay in formalin-fixed paraffin sections of human breast cancer tissue using monoclonal antibodies. Br J Cancer. 1986 May;53(5):691–694. doi: 10.1038/bjc.1986.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benraad T., Koenders A. Estradiol receptor activity in lyophilized calf uterus and human breast tumor tissue. Cancer. 1980 Dec 15;46(12 Suppl):2762–2764. doi: 10.1002/1097-0142(19801215)46:12+<2762::aid-cncr2820461403>3.0.co;2-o. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Davis B. W., Zava D. T., Locher G. W., Goldhirsch A., Hartmann W. H. Receptor heterogeneity of human breast cancer as measured by multiple intratumoral assays of estrogen and progesterone receptor. Eur J Cancer Clin Oncol. 1984 Mar;20(3):375–382. doi: 10.1016/0277-5379(84)90084-1. [DOI] [PubMed] [Google Scholar]
  6. De Goeij A. F., Volleberg M. P., Hondius G. G., Bosman F. T. Radiochemical determination of estrogen receptors in cryostat sections of target tissues. J Steroid Biochem. 1984 Aug;21(2):127–134. doi: 10.1016/0022-4731(84)90372-8. [DOI] [PubMed] [Google Scholar]
  7. De Goeij T. F., Bosman F. T., Berns E. M. Determination of steroid hormone-dependency of tumours utilizing tissue sections. Survey of histochemical techniques and their application in surgical pathology. J Pathol. 1986 Jul;149(3):163–172. doi: 10.1002/path.1711490302. [DOI] [PubMed] [Google Scholar]
  8. DeSombre E. R., Thorpe S. M., Rose C., Blough R. R., Andersen K. W., Rasmussen B. B., King W. J. Prognostic usefulness of estrogen receptor immunocytochemical assays for human breast cancer. Cancer Res. 1986 Aug;46(8 Suppl):4256s–4264s. [PubMed] [Google Scholar]
  9. Geyer H., Eberl M., Hornberger B. Influence of tissue heterogeneity on the determination of steroid receptors in breast cancer. J Cancer Res Clin Oncol. 1985;110(2):141–144. doi: 10.1007/BF00402728. [DOI] [PubMed] [Google Scholar]
  10. Giri D. D., Dangerfield V. J., Lonsdale R., Rogers K., Underwood J. C. Immunohistology of oestrogen receptor and D5 antigen in breast cancer: correlation with oestrogen receptor content of adjacent cryostat sections assayed by radioligand binding and enzyme immunoassay. J Clin Pathol. 1987 Jul;40(7):734–740. doi: 10.1136/jcp.40.7.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Giri D. D., Lonsdale R. N., Dangerfield V. J., Harris S. C., Parsons M. A., Underwood J. C. Clinicopathological significance of intratumoural variations in elastosis grades and the oestrogen receptor status of human breast carcinomas. J Pathol. 1987 Apr;151(4):297–303. doi: 10.1002/path.1711510410. [DOI] [PubMed] [Google Scholar]
  12. Greene G. L., Sobel N. B., King W. J., Jensen E. V. Immunochemical studies of estrogen receptors. J Steroid Biochem. 1984 Jan;20(1):51–56. doi: 10.1016/0022-4731(84)90188-2. [DOI] [PubMed] [Google Scholar]
  13. Hawkins R. A., Sangster K., Krajewski A. Histochemical detection of oestrogen receptors in breast carcinoma: a successful technique. Br J Cancer. 1986 Mar;53(3):407–410. doi: 10.1038/bjc.1986.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heubner A., Beck T., Grill H. J., Pollow K. Comparison of immunocytochemical estrogen receptor assay, estrogen receptor enzyme immunoassay, and radioligand-labeled estrogen receptor assay in human breast cancer and uterine tissue. Cancer Res. 1986 Aug;46(8 Suppl):4291s–4295s. [PubMed] [Google Scholar]
  15. Howanitz P. J., Howanitz J. H., Skrodzki C. A., Woloszyn T. Protein method influences on calculation of tissue receptor concentration. Am J Clin Pathol. 1986 Jan;85(1):37–42. doi: 10.1093/ajcp/85.1.37. [DOI] [PubMed] [Google Scholar]
  16. Hull D. F., 3rd, Clark G. M., Osborne C. K., Chamness G. C., Knight W. A., 3rd, McGuire W. L. Multiple estrogen receptor assays in human breast cancer. Cancer Res. 1983 Jan;43(1):413–416. [PubMed] [Google Scholar]
  17. Jonat W., Maass H., Stegner H. E. Immunohistochemical measurement of estrogen receptors in breast cancer tissue samples. Cancer Res. 1986 Aug;46(8 Suppl):4296s–4298s. [PubMed] [Google Scholar]
  18. Jordan V. C., Zava D. T., Eppenburger U., Kiser A., Sebek S., Dowdle E., Krozowski Z., Bennett R. C., Funder J., Holdaway I. M. Reliability of steroid hormone receptor assays: an international study. Eur J Cancer Clin Oncol. 1983 Mar;19(3):357–363. doi: 10.1016/0277-5379(83)90133-5. [DOI] [PubMed] [Google Scholar]
  19. King W. J., DeSombre E. R., Jensen E. V., Greene G. L. Comparison of immunocytochemical and steroid-binding assays for estrogen receptor in human breast tumors. Cancer Res. 1985 Jan;45(1):293–304. [PubMed] [Google Scholar]
  20. King W. J., Greene G. L. Monoclonal antibodies localize oestrogen receptor in the nuclei of target cells. Nature. 1984 Feb 23;307(5953):745–747. doi: 10.1038/307745a0. [DOI] [PubMed] [Google Scholar]
  21. Koenders A., Thorpe S. M. Standardization of steroid receptor assays in human breast cancer--IV. Long-term within- and between-laboratory variation of estrogen and progesterone receptor assays. Eur J Cancer Clin Oncol. 1986 Aug;22(8):945–952. doi: 10.1016/0277-5379(86)90061-1. [DOI] [PubMed] [Google Scholar]
  22. Leclercq G., Bojar H., Goussard J., Nicholson R. I., Pichon M. F., Piffanelli A., Pousette A., Thorpe S., Lonsdorfer M. Abbott monoclonal enzyme immunoassay measurement of estrogen receptors in human breast cancer: a European multicenter study. Cancer Res. 1986 Aug;46(8 Suppl):4233s–4236s. [PubMed] [Google Scholar]
  23. McCarty K. S., Jr, Miller L. S., Cox E. B., Konrath J., McCarty K. S., Sr Estrogen receptor analyses. Correlation of biochemical and immunohistochemical methods using monoclonal antireceptor antibodies. Arch Pathol Lab Med. 1985 Aug;109(8):716–721. [PubMed] [Google Scholar]
  24. Norgren A., Fernö M., Borg A. Observations on wet weight, protein and DNA as reference for steroid receptors in malignant mammary tumours. Anticancer Res. 1986 Jan-Feb;6(1):59–64. [PubMed] [Google Scholar]
  25. Osborne C. K. Heterogeneity in hormone receptor status in primary and metastatic breast cancer. Semin Oncol. 1985 Sep;12(3):317–326. [PubMed] [Google Scholar]
  26. Press M. F., Greene G. L. An immunocytochemical method for demonstrating estrogen receptor in human uterus using monoclonal antibodies to human estrophilin. Lab Invest. 1984 Apr;50(4):480–486. [PubMed] [Google Scholar]
  27. Shimada A., Kimura S., Abe K., Nagasaki K., Adachi I., Yamaguchi K., Suzuki M., Nakajima T., Miller L. S. Immunocytochemical staining of estrogen receptor in paraffin sections of human breast cancer by use of monoclonal antibody: comparison with that in frozen sections. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4803–4807. doi: 10.1073/pnas.82.14.4803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shintaku I. P., Said J. W. Detection of estrogen receptors with monoclonal antibodies in routinely processed formalin-fixed paraffin sections of breast carcinoma. Use of DNase pretreatment to enhance sensitivity of the reaction. Am J Clin Pathol. 1987 Feb;87(2):161–167. doi: 10.1093/ajcp/87.2.161. [DOI] [PubMed] [Google Scholar]
  29. Silfverswärd C., Skoog L., Humla S., Gustafsson S. A., Nordenskjöld B. Intratumoral variation of cytoplasmic and nuclear estrogen receptor concentrations in human mammary carcinoma. Eur J Cancer. 1980 Jan;16(1):59–65. doi: 10.1016/0014-2964(80)90108-5. [DOI] [PubMed] [Google Scholar]
  30. Stefanini M., De Martino C., Zamboni L. Fixation of ejaculated spermatozoa for electron microscopy. Nature. 1967 Oct 14;216(5111):173–174. doi: 10.1038/216173a0. [DOI] [PubMed] [Google Scholar]
  31. Teasdale J., Jackson P., Holgate C. S., Cowen P. N. Identification of oestrogen receptors in cells of paraffin-processed breast cancers by IGSS. Histochemistry. 1987;87(2):185–187. doi: 10.1007/BF00533404. [DOI] [PubMed] [Google Scholar]
  32. Thorpe S. M., Koenders A. Standardization of steroid receptor assays in human breast cancer--III. Selection of reference material for intra- and inter-laboratory quality control. Eur J Cancer Clin Oncol. 1986 Aug;22(8):939–944. doi: 10.1016/0277-5379(86)90060-x. [DOI] [PubMed] [Google Scholar]
  33. Underwood J. C., Dangerfield V. J., Parsons M. A. Oestrogen receptor assay of cryostat sections of human breast carcinomas with simultaneous quantitative histology. J Clin Pathol. 1983 Apr;36(4):399–405. doi: 10.1136/jcp.36.4.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Underwood J. C. Oestrogen receptors in human breast cancer: review of histopathological correlations and critique of histochemical methods. Diagn Histopathol. 1983 Jan-Mar;6(1):1–22. [PubMed] [Google Scholar]
  35. van Netten J. P., Algard F. T., Coy P., Carlyle S. J., Brigden M. L., Thornton K. R., Peter S., Fraser T., To M. P. Heterogeneous estrogen receptor levels detected via multiple microsamples from individual breast cancers. Cancer. 1985 Oct 15;56(8):2019–2024. doi: 10.1002/1097-0142(19851015)56:8<2019::aid-cncr2820560822>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES