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. 1994 Jun;69(6):1032–1037. doi: 10.1038/bjc.1994.203

Dual immunocytochemical analysis of oestrogen and epidermal growth factor receptors in human breast cancer.

A K Sharma 1, K Horgan 1, A Douglas-Jones 1, R McClelland 1, J Gee 1, R Nicholson 1
PMCID: PMC1969412  PMID: 8198966

Abstract

Recent studies have demonstrated a consistent inverse relationship between oestrogen receptor (ER) and epidermal growth factor receptor (EGFR) levels in female human breast cancer. Serial cross-section studies have suggested that separate populations of ER+/EGFR- and ER-/EGFR+ cancer cells exist in tumours deemed by immunocytochemical assay (ICA) to be positive for both. We have developed a dual ICA that is able to stain for both ER and EGFR on a single 5 microns frozen section sample of breast tissue. Twenty-two samples of female human breast cancer tissue that exhibited positivity for ER and EGFR by ER-ICA using the H222 monoclonal antibody and EGFR-ICA using the EGFR1 monoclonal antibody underwent the dual ICA. There was a significant correlation in receptor positivity between the single and dual assays for both ER (rs = 0.801, P < 0.001) and EGFR (rs = 0.831, P < 0.001). Individual cancer cells exhibited one of three staining patterns: nuclear staining only (ER+/EGFR-), membrane-associated and cytoplasmic staining only (ER-/EGFR+) or no staining (ER-/EGFR-). No cancer cells exhibited both nuclear and membrane/cytoplasmic staining. This is the first description of a simultaneous dual immunocytochemical assay system for ER and EGFR in clinical breast cancer specimens. The results suggest that ER and EGFR expression are mutually exclusive within an individual breast cancer cell in vivo with separate populations of ER+/EGFR- cells, ER-/EGFR+ cells and ER-/EGFR- cells coexisting.

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Selected References

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

  1. Bevilacqua P., Gasparini G., Dal Fior S., Corradi G. Immunocytochemical determination of epidermal growth factor receptor with monoclonal EGFR1 antibody in primary breast cancer patients. Oncology. 1990;47(4):313–317. doi: 10.1159/000226840. [DOI] [PubMed] [Google Scholar]
  2. Bilous M., Milliken J., Mathijs J. M. Immunocytochemistry and in situ hybridisation of epidermal growth factor receptor and relation to prognostic factors in breast cancer. Eur J Cancer. 1992;28A(6-7):1033–1037. doi: 10.1016/0959-8049(92)90449-c. [DOI] [PubMed] [Google Scholar]
  3. Bolufer P., Miralles F., Rodriguez A., Vazquez C., Lluch A., Garcia-Conde J., Olmos T. Epidermal growth factor receptor in human breast cancer: correlation with cytosolic and nuclear ER receptors and with biological and histological tumor characteristics. Eur J Cancer. 1990 Mar;26(3):283–290. doi: 10.1016/0277-5379(90)90223-g. [DOI] [PubMed] [Google Scholar]
  4. Cohen S., Ushiro H., Stoscheck C., Chinkers M. A native 170,000 epidermal growth factor receptor-kinase complex from shed plasma membrane vesicles. J Biol Chem. 1982 Feb 10;257(3):1523–1531. [PubMed] [Google Scholar]
  5. De Larco J. E., Todaro G. J. Sarcoma growth factor (SGF): specific binding to epidermal growth factor (EGF) membrane receptors. J Cell Physiol. 1980 Feb;102(2):267–277. doi: 10.1002/jcp.1041020218. [DOI] [PubMed] [Google Scholar]
  6. Downward J., Parker P., Waterfield M. D. Autophosphorylation sites on the epidermal growth factor receptor. Nature. 1984 Oct 4;311(5985):483–485. doi: 10.1038/311483a0. [DOI] [PubMed] [Google Scholar]
  7. Ewing T. M., Murphy L. J., Ng M. L., Pang G. Y., Lee C. S., Watts C. K., Sutherland R. L. Regulation of epidermal growth factor receptor by progestins and glucocorticoids in human breast cancer cell lines. Int J Cancer. 1989 Oct 15;44(4):744–752. doi: 10.1002/ijc.2910440432. [DOI] [PubMed] [Google Scholar]
  8. Gorski J., Welshons W. V., Sakai D., Hansen J., Walent J., Kassis J., Shull J., Stack G., Campen C. Evolution of a model of estrogen action. Recent Prog Horm Res. 1986;42:297–329. doi: 10.1016/b978-0-12-571142-5.50011-2. [DOI] [PubMed] [Google Scholar]
  9. Hainsworth P. J., Henderson M. A., Stillwell R. G., Bennett R. C. Comparison of EGFR, c-erbB-2 product and ras p21 immunohistochemistry as prognostic markers in primary breast cancer. Eur J Surg Oncol. 1991 Feb;17(1):9–15. [PubMed] [Google Scholar]
  10. Horwitz K. B., Wei L. L., Sedlacek S. M., d'Arville C. N. Progestin action and progesterone receptor structure in human breast cancer: a review. Recent Prog Horm Res. 1985;41:249–316. doi: 10.1016/b978-0-12-571141-8.50010-x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Klijn J. G., Berns P. M., Schmitz P. I., Foekens J. A. The clinical significance of epidermal growth factor receptor (EGF-R) in human breast cancer: a review on 5232 patients. Endocr Rev. 1992 Feb;13(1):3–17. doi: 10.1210/edrv-13-1-3. [DOI] [PubMed] [Google Scholar]
  14. Lee C. S., Hall R. E., Alexander I. E., Koga M., Shine J., Sutherland R. L. Inverse relationship between estrogen receptor and epidermal growth factor receptor mRNA levels in human breast cancer cell lines. Growth Factors. 1990;3(2):97–103. doi: 10.3109/08977199009108272. [DOI] [PubMed] [Google Scholar]
  15. Lee C. S., Koga M., Sutherland R. L. Modulation of estrogen receptor and epidermal growth factor receptor mRNAs by phorbol ester in MCF 7 breast cancer cells. Biochem Biophys Res Commun. 1989 Jul 14;162(1):415–421. doi: 10.1016/0006-291x(89)92013-5. [DOI] [PubMed] [Google Scholar]
  16. Lewis S., Locker A., Todd J. H., Bell J. A., Nicholson R., Elston C. W., Blamey R. W., Ellis I. O. Expression of epidermal growth factor receptor in breast carcinoma. J Clin Pathol. 1990 May;43(5):385–389. doi: 10.1136/jcp.43.5.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lippman M. E., Dickson R. B., Gelmann E. P., Rosen N., Knabbe C., Bates S., Bronzert D., Huff K., Kasid A. Growth regulation of human breast carcinoma occurs through regulated growth factor secretion. J Cell Biochem. 1987 Sep;35(1):1–16. doi: 10.1002/jcb.240350102. [DOI] [PubMed] [Google Scholar]
  18. Lippman M. E., Dickson R. B. Mechanisms of growth control in normal and malignant breast epithelium. Recent Prog Horm Res. 1989;45:383–440. doi: 10.1016/b978-0-12-571145-6.50012-1. [DOI] [PubMed] [Google Scholar]
  19. McClelland R. A., Berger U., Miller L. S., Powles T. J., Coombes R. C. Immunocytochemical assay for estrogen receptor in patients with breast cancer: relationship to a biochemical assay and to outcome of therapy. J Clin Oncol. 1986 Aug;4(8):1171–1176. doi: 10.1200/JCO.1986.4.8.1171. [DOI] [PubMed] [Google Scholar]
  20. McClelland R. A., Wilson D., Leake R., Finlay P., Nicholson R. I. A multicentre study into the reliability of steroid receptor immunocytochemical assay quantification. British Quality Control Group. Eur J Cancer. 1991;27(6):711–715. doi: 10.1016/0277-5379(91)90171-9. [DOI] [PubMed] [Google Scholar]
  21. Nicholson S., Wright C., Sainsbury J. R., Halcrow P., Kelly P., Angus B., Farndon J. R., Harris A. L. Epidermal growth factor receptor (EGFr) as a marker for poor prognosis in node-negative breast cancer patients: neu and tamoxifen failure. J Steroid Biochem Mol Biol. 1990 Dec 20;37(6):811–814. doi: 10.1016/0960-0760(90)90424-j. [DOI] [PubMed] [Google Scholar]
  22. Ro J., North S. M., Gallick G. E., Hortobagyi G. N., Gutterman J. U., Blick M. Amplified and overexpressed epidermal growth factor receptor gene in uncultured primary human breast carcinoma. Cancer Res. 1988 Jan 1;48(1):161–164. [PubMed] [Google Scholar]
  23. Roberts A. B., Frolik C. A., Anzano M. A., Sporn M. B. Transforming growth factors from neoplastic and nonneoplastic tissues. Fed Proc. 1983 Jun;42(9):2621–2626. [PubMed] [Google Scholar]
  24. Sainsbury J. R., Farndon J. R., Sherbet G. V., Harris A. L. Epidermal-growth-factor receptors and oestrogen receptors in human breast cancer. Lancet. 1985 Feb 16;1(8425):364–366. doi: 10.1016/s0140-6736(85)91385-6. [DOI] [PubMed] [Google Scholar]
  25. Toi M., Hamada Y., Nakamura T., Mukaida H., Suehiro S., Wada T., Toge T., Niimoto M., Hattori T. Immunocytochemical and biochemical analysis of epidermal growth factor receptor expression in human breast cancer tissues: relationship to estrogen receptor and lymphatic invasion. Int J Cancer. 1989 Feb 15;43(2):220–225. doi: 10.1002/ijc.2910430208. [DOI] [PubMed] [Google Scholar]
  26. Toi M., Nakamura T., Mukaida H., Wada T., Osaki A., Yamada H., Toge T., Niimoto M., Hattori T. Relationship between epidermal growth factor receptor status and various prognostic factors in human breast cancer. Cancer. 1990 May 1;65(9):1980–1984. doi: 10.1002/1097-0142(19900501)65:9<1980::aid-cncr2820650917>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  27. Travers M. T., Barrett-Lee P. J., Berger U., Luqmani Y. A., Gazet J. C., Powles T. J., Coombes R. C. Growth factor expression in normal, benign, and malignant breast tissue. Br Med J (Clin Res Ed) 1988 Jun 11;296(6637):1621–1624. doi: 10.1136/bmj.296.6637.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Walker K. J., Bouzubar N., Robertson J., Ellis I. O., Elston C. W., Blamey R. W., Wilson D. W., Griffiths K., Nicholson R. I. Immunocytochemical localization of estrogen receptor in human breast tissue. Cancer Res. 1988 Nov 15;48(22):6517–6522. [PubMed] [Google Scholar]
  29. Waterfield M. D., Mayes E. L., Stroobant P., Bennet P. L., Young S., Goodfellow P. N., Banting G. S., Ozanne B. A monoclonal antibody to the human epidermal growth factor receptor. J Cell Biochem. 1982;20(2):149–161. doi: 10.1002/jcb.240200207. [DOI] [PubMed] [Google Scholar]
  30. Watts C. K., Handel M. L., King R. J., Sutherland R. L. Oestrogen receptor gene structure and function in breast cancer. J Steroid Biochem Mol Biol. 1992 Mar;41(3-8):529–536. doi: 10.1016/0960-0760(92)90378-v. [DOI] [PubMed] [Google Scholar]

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