Skip to main content
SpringerLink
Log in

Tamoxifen aziridine labeling of the estrogen receptor — potential utility in detecting biologically aggressive breast tumors

  • Report
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Summary

Expression of estrogen receptor (ER) is a helpful predictor of response to endocrine therapy and disease free survival in breast cancer patients. The presence of variant estrogen receptors has been demonstrated at the RNA/DNA level and might represent an escape of tumors from hormonal control mechanisms. However, the demonstration that the corresponding peptides do exist is a real challenge. Denaturing polyacrylamide gel electrophoresis (SDS-PAGE) of covalently bound [3H]tamoxifen aziridine ([3H]TAZ) to ER demonstrates a specific, multiband peptide pattern recognized by anti-ER monoclonal antibodies (anti-ER Mo Abs). The native 66 kDa ER form identified through its hormone binding domain by the H-222 Mo Ab was the most prominent one followed by 50, 35, and 28 kDa forms on fluorography. Such patterns from early human breast tumors were compared to the ones of more advanced disease, namely large primary breast cancers, metastatic lymph nodes, and soft tissue relapses: in these cases, molecular forms of 43 and 35 kDa were identified with a remarkable consistency. The 43 kDa peptide was more frequently identified by the H-226 Mo Ab (which maps a region near the DNA binding domain) — albeit with low labeling intensity as compared to H-222 Mo Ab. In addition, the 43 kDa peptide was inversely correlated to ER levels. This altered ER or related peptide could potentially be a marker of biologically aggressive breast tumors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  1. McGuire WL: Hormone receptors: Their role in predicting prognosis and response to endocrine therapy. Semin Oncol 5: 428–433, 1978

    PubMed  Google Scholar 

  2. Green GL, Nolan C, Engler JP, Jensen EV: Monoclonal antibodies to human estrogen receptor. Proc Natl Acad Sci USA 77: 5115–5110, 1980

    PubMed  Google Scholar 

  3. Kumar V, Green S, Stack G, Berry M, Jin JR, Chambon P: Functional domains of the human estrogen receptor. Cell 51: 941–951, 1987

    PubMed  Google Scholar 

  4. Fuqua SAW, Fitzgerald SD, Allred DC, Elledge RM, Nawaz Z, McDonnell DP, O'Malley BW, Green GL, McGuire WL: Inhibition of estrogen receptor action by a naturally occurring variant in human breast tumors. Cancer Res 52: 483–486, 1992

    PubMed  Google Scholar 

  5. Castles C, Fuqua SAW, Klotz DM, Hill SM: Expression of a constitutively active estrogen receptor variant in the estrogen receptor negative BT-20 human breast cancer cell line. Cancer Res 53: 5934–5939, 1993

    PubMed  Google Scholar 

  6. Pfeffer U, Fecarotta E, Castagnetta L, Vidali G: Estrogen receptor variant messager RNA lacking exon 4 in estrogenresponsive human breast cancer cell lines. Cancer Res 53: 741–743, 1993

    PubMed  Google Scholar 

  7. McGuire WL, Chamness GC, Fuqua SAW: Estrogen receptor variants in clinical breast cancer. Mol Endocrinol 5: 1571–1577, 1991

    PubMed  Google Scholar 

  8. Garcia T, Lehrer S, Bloomer WD, Schachter B: A variant estrogen receptor messenger ribonucleic acid is associated with reduced levels of estrogen binding in human mammary tissues. Mol Endocrinol 2: 785–791, 1988

    PubMed  Google Scholar 

  9. Dotzlaw H, Alkhalaf M, Murphy LC: Characterization of estrogen receptor variant mRNAs from human breast cancer. Mol Endocrinol 6: 773–785, 1992

    PubMed  Google Scholar 

  10. Murphy LC, Hilsenbeck SG, Dotzlaw H, Fuqua SAW: Relationship of clone 4 estrogen receptor variant messenger RNA expression to some known prognostic variables in human breast cancer. Clinical Can Res 1: 155–159, 1995

    Google Scholar 

  11. Piccart MJ, Muquardt C, Bosman C, Pirotte P, Veenstra S, Grillo F, Leclercq G: Comparison of tritiated estradiol and tamoxifen aziridine for measurement of estrogen receptors in human breast cancer cytosols. J Natl Cancer Inst 83: 1553–1559, 1991

    PubMed  Google Scholar 

  12. Giambiagi N, Pasqualini JR: Transformation of the estrogen receptor detected by two monoclonal antibodies. J Steroid Biochem 34: 297–300, 1989

    PubMed  Google Scholar 

  13. European Organization for Research and Treatment of Cancer: Breast Cancer Co-operative Group. Revision of the standards for the assessment of hormone receptors in human breast cancer: report of the second E.O.R.T.C. Workshop, held March 16–17 in The Netherlands Cancer Institute. Eur J Cancer 16: 1513–1515, 1980

    Google Scholar 

  14. Horigome T, Ogata F, Golding TS, Korach KS: Estradiol stimulated proteolytic cleavage of the estrogen receptor in mouse uterus. Endocrinology 123: 2540–2548, 1988

    PubMed  Google Scholar 

  15. Maaroufi Y, Trivedi S, Leclercq G: Major molecular weight heterogeneity of estrogen receptor from breast cancer is not related to neoplasia. Cancer Biochem Biophys 15: 67–78, 1995

    PubMed  Google Scholar 

  16. Maaroufi Y, Leclercq G: Importance of A/B and C domains of the estrogen receptor for its adsorption to hydroxylapatite. J Steroid Biochem Mol Biol 48: 155–163, 1994

    PubMed  Google Scholar 

  17. Salomonsson M, Häggblad J, O'Malley BW, Sitbon GM: The human estrogen receptor hormone binding domain dimerizes independently of ligand activator. J Steroid Biochem Mol Biol 48: 447–452, 1994

    PubMed  Google Scholar 

  18. McCague R, Potter KA: Synthesis of 4-stannylated tamoxifen analogues; useful precursors to radiolabeled iodoxifene and aziridinyl 4-iodotamoxifen. J Lab Comp Radiopharm 34: 297–302, 1994

    Google Scholar 

  19. Rees JC, Wooge CH, Katzenellenbogen BS: Identification of two cysteines closely positioned in the ligand binding pocket of the human estrogen receptor: Roles in ligand binding and transcriptional activation. Mol Endocrinol 6: 2160–2166, 1992

    PubMed  Google Scholar 

  20. Toulas C, Barraille P, Delassus F, Bayard F, Prats H, Faye JC: Differential initiation of translation of a single estrogen receptor mRNA could explain some estradiol resistance cases. Horm Res 38: 73–77, 1992

    Google Scholar 

  21. Sluyser M: Role of estrogen receptor variants in the development of hormone resistance in breast cancer. Clin Biochem 25: 407–414, 1992

    PubMed  Google Scholar 

  22. Pakdel F, Katzenellenbogen BS: Human estrogen receptor mutants with altered estrogen and antiestrogen ligand discrimination. J Biol Chem 267: 3429–3437, 1992

    PubMed  Google Scholar 

  23. Danielian PS, White R, Hoare SA, Fawell SE, Parker MG: Identification of residues in the estrogen receptor that confer differential sensitivity to estrogen and hydroxytamoxifen. Mol Endocrinol 7: 232–240, 1993

    PubMed  Google Scholar 

  24. Borgna JL, Scali J: Differential inhibition of estrogen and antiestrogen binding to the estrogen receptor by diethylpyrocarbonate. J Steroid Biochem 31: 427–436, 1988

    PubMed  Google Scholar 

  25. Borgna JL, Scali J: Differential interactions of estrogens and antiestrogens at the 17β-hydroxy or counterpart function with the estrogen receptor. Eur J Biochem 199: 575–585, 1991

    PubMed  Google Scholar 

  26. Koeherst SKA, Jacobs HM, Thijssen JHH, Blankenstein MA: Detection of an oestrogen receptor-like protein in human meningiomas by band shift assay using a synthetic oestrogen responsive element (ERE). Br J Cancer 68: 290–294, 1993

    PubMed  Google Scholar 

  27. Sluyser M: Mutations in the estrogen receptor gene. Hum Mutations 6: 97–103, 1995

    Google Scholar 

  28. Messeri G, Tozzi P, Quercioli M, Cataliotti L, Cardona G: Alkaline phosphatases and steroid receptors in human breast cancer. J Steroid Biochem Mol Biol 37: 269–271, 1990

    PubMed  Google Scholar 

  29. Auricchio F: Phosphorylation of steroid receptors: General review. J Steroid Biochem 32: 613–622, 1989

    PubMed  Google Scholar 

  30. McNaught RW, Dayani N, Smith RG: Receptor interconversion model of hormone action. I. Purification of a factor involved in conferring estradiol binding to the estrogen receptor. Biochem 29: 2685–2690, 1990

    Google Scholar 

  31. Migliaccio A, Pagano M, De Goeij CCJ, Di Domenico M, Castoria G, Sluyser M, Auricchio F: Phosphorylation and estradiol binding of estrogen receptor in hormone-dependent and hormone-independent GR mouse mammary tumors. Int J Cancer 51: 733–739, 1992

    PubMed  Google Scholar 

  32. Graham ML, Krett NL, Miller LA, Leslie KK, Gordon DF, Wood WM, Wei LL, Horwitz KB: T47Dco cells, genetically unstable and containing estrogen receptor mutations, are a model for the progression of breast cancers to hormone resistance. Cancer Res 50: 6208–6217, 1990

    PubMed  Google Scholar 

  33. Fuqua SAW, Falette NF, McGuire WL: Sensitive detection of estrogen receptor RNA by polymerase chain reaction assay. J Natl Cancer Inst 82: 858–861, 1990

    PubMed  Google Scholar 

  34. Murphy LC: Estrogen receptor variants in human breast cancer. Mol Cell Endocrinol 74: C83-C86, 1990

    PubMed  Google Scholar 

  35. Castles CG, Klotz DM, Fuqua SAW, Hill SM: Coexpression of wild-type and variant oestrogen receptor mRNAs in a panel of human breast cancer cell lines. Br J Cancer 71: 974–980, 1995

    PubMed  Google Scholar 

  36. Foster BD, Cavener DR, Parl FF: Binding analysis of the estrogen receptor to its specific DNA target site in human breast cancer. Cancer Res 51: 3405–3410, 1991

    PubMed  Google Scholar 

  37. Scott GK, Kushner P, Vigne J, Benz CC: Truncated forms of DNA-binding estrogen receptors in human breast cancer. J Clin Invest 88: 700–706, 1991

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Sunil Trivedi

    Present address: Division of Molecular Endocrinology, The Gujarat Cancer and Research Institute, 380 016, Ahmedabad, India

Authors and Affiliations

  1. Laboratoire J.-C. Heuson de Cancérologie Mammaire, Service de Médecine, Institut Jules Bordet, Rue Heger Bordet 1, 1000, Brussels, Belgium

    Sunil Trivedi, Carl Muquardt, Nathalie Gilot, Samira Hadiy & Guy Leclercq

  2. Unité de Chimiothérapie, Service de Médecine, Institut Jules Bordet, Rue Heger Bordet 1, 1000, Brussels, Belgium

    Martine Piccart

  3. Surgical Specialities, 380 016, Ahmedabad, India

    Devendra Patel (Coordinator, Director)

  4. Division of Molecular Endocrinology, The Gujarat Cancer and Research Institute, 380 016, Ahmedabad, India

    Devendra Patel (Coordinator, Director)

Authors
  1. Sunil Trivedi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Martine Piccart
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carl Muquardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nathalie Gilot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Samira Hadiy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Devendra Patel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guy Leclercq
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Trivedi, S., Piccart, M., Muquardt, C. et al. Tamoxifen aziridine labeling of the estrogen receptor — potential utility in detecting biologically aggressive breast tumors. Breast Cancer Res Tr 40, 231–241 (1996). https://doi.org/10.1007/BF01806811

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01806811

Key words

Search

Navigation