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Influence of estrogen receptor variants on the determination of ER status in human breast cancer

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Abstract

Determination of estrogen receptor alpha (ER) status in breast cancer is an important predictive factor for clinical response to endocrine therapy. We have recently shown that discrepancies in ER status determined by immunohistochemical assay (ER-IHA) can occur between amino-terminal (1D5) and carboxyl-terminal (AER-311) targeted ER antibodies and that those tumors which demonstrate discordance are associated with increased expression of truncated ER variant mRNAs. In this study, we have explored this observation to examine if ER variant expression can exert a direct effect on ER-IHA or whether this association is attributable to the characteristics of the antibodies. ER negative cos-1 cells were transfected with expression vectors containing wild type ER (wt-ER) and/or a frequently expressed truncated variant, ER-clone-4 variant. We found that ER-IHA performed with the same N- and C-terminal targeting ER antibodies on cos-1 cells expressing wt-ER alone demonstrated no difference in signals by western blot (P>0.1). However, co-expression of wt-ER and the truncated ER-clone-4 variant, resulted in discordant IHA results with relatively higher ER-IHA H-scores from N-terminal antibodies (P<0.03). Furthermore, re-examination of a subset of breast tumors previously studied by ER-IHA showed persistent concordance in 4/5 cases and persistent differences in 3/5 cases with a different pair of ER antibodies. We conclude that the presence of truncated ER variant proteins can interfere with the interpretation of ER status determined by IHA and that this may account for some of the inconsistencies between ER status and response to endocrine therapy.

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References

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

    Google Scholar 

  2. Dotzlaw H, Leygue E, Watson PH, Murphy LC: Expression of estrogen receptor-beta in human breast tumors. J Clin Endocrinol Metab 82: 2371–2374, 1997

    Google Scholar 

  3. Murphy LC, Leygue E, Dotzlaw H, Douglas D, Coutts A, Watson PH: Oestrogen receptor variants and mutations in human breast cancer. Ann Med 29: 221–234, 1997

    Google Scholar 

  4. Leygue E, Huang A, Murphy L, Watson P: Prevalence of estrogen receptor variant mRNAs in human breast cancer. Cancer Res 56: 4324–4327, 1996

    Google Scholar 

  5. Park W, Choi JJ, Hwang ES, Lee JH: Identification of a variant estrogen receptor lacking exon 4 and its coexpression with wild-type estrogen receptor in ovarian carcinomas. Clin Cancer Res 2: 2029–2035, 1996

    Google Scholar 

  6. Desai AJ, Luqmani YA, Walters JE, Coope RC, Dagg B, Gomm JJ, Pace PE, Rees CN, Thirunavukkarasu V, Shousha S, Groome NP, Coombes R, Ali S: Presence of exon 5-deleted oestrogen receptor in human breast cancer: functional analysis and clinical significance. Brit J Cancer 75: 1173–1184, 1997

    Google Scholar 

  7. Taylor CR.: Paraffin section immunocytochemistry for estrogen receptor. Editorial Cancer 77: 2419–2422, 1996

    Google Scholar 

  8. Allred DC, Bustamante MA, Daniel CO, Gaskill HV, Cruz AJ: Immunocytochemical analysis of estrogen receptor in human breast carcinomas. Evaluation of 130 cases and review of the literature regarding concordance with biochemical assay and clinical relevance. Arch Surg 125: 107–113, 1990

    Google Scholar 

  9. Alberts SR, Ingle JN, Roche PR, Cha SS, Wold LE, Farr GH, Krook JE, Wieland HS: Comparison of estrogen receptor determinations by a biochemical ligand assay and immunohistochemical staining with monoclonal antibody ER1D5 in females with lymph node positive breast carcinoma entered on two prospective clinical trials. Cancer 78: 764–772, 1996

    Google Scholar 

  10. Dowsett M, Daffada A, Chan CMW, Johnston SRD: Oestrogen receptor mutants and variants in breast cancer. Euro J Cancer 33: 1177–1183, 1997

    Google Scholar 

  11. Huang A, Pettigrew N, Watson P: Immunohistochemical assay for estrogen receptors in paraffin wax sections of breast carcinoma using a new monoclonal antibody. J Pathol 180: 223–227, 1996

    Google Scholar 

  12. Pertschuk LP, Feldman JG, Kim YD, Braithwaite L, Schneider F, Braverman AS, Axiotis C: Estrogen receptor immunocytochemistry in paraffin embedded tissues with ER1D5 predicts breast cancer endocrine response more accurately than H222Spg in frozen sections or cytosol-based ligandbinding assays. Cancer 77: 2514–2519, 1996

    Google Scholar 

  13. Nedergaard L, Christensen L, Rasmussen BB, Jacobsen GK: Comparison of two monoclonal antibodies for the detection of estrogen receptors in primary breast carcinomas. Pathol Res Pract 192: 983–988, 1996

    Google Scholar 

  14. Huang A, Leygue E, Snell L, Murphy L, Watson P: Expression of estrogen receptor variant mRNAs and determination of estrogen receptor status in human breast cancer. Am J Pathol 150: 1827–1833, 1997

    Google Scholar 

  15. Dotzlaw H, Alkhalaf M, Murphy L: Characterization of estrogen receptor variant mRNAs from breast cancers. Mol Endocrinol 6: 773–785, 1992

    Google Scholar 

  16. 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. Clin Cancer Res 1: 155–159, 1995

    Google Scholar 

  17. Pfeffer U, Fecarotta E, Arena G, Forlani A, Vidali G: Alternate splicing of the estrogen receptor primary transcript normally occurs in estrogen receptor positive tissues and cell lines. J Steroid Biochem 56: 99–105, 1996

    Google Scholar 

  18. Gotteland M, Desauty G, Delarue JC, Liu L, May E: Human estrogen receptor messenger RNA variants in both normal and tumor tissues. Mol Cell Endocrinol 112: 1–13, 1995

    Google Scholar 

  19. Leygue E, Watson P, Murphy L: Estrogen receptor variants in normal human mammary tissue. J Natl Cancer Inst 88: 284–290, 1996

    Google Scholar 

  20. Leygue E, Murphy L, Watson P: Triple primer polymerase chain reaction: a new way to quantify truncated mRNA expression. Am J Pathol 148: 1097–1103, 1996

    Google Scholar 

  21. Castles CG, 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

    Google Scholar 

  22. Pink JJ, Wu SQ, Wolf DM, Bilimoria MM, Jordan VC: A novel 80 kDa human estrogen receptor containing a duplication of exons 6 and 7. Nucleic Acids Res 4: 962–969, 1996

    Google Scholar 

  23. Madsen MW, Reiter BE, Lykkesfeldt AE: Differential expression of estrogen receptor mRNA splice variants in the tamoxifen resistant human breast cancer cell line, MCF-7/TAMR-1 compared to the parental MCF-7 cell line. Mol Cell Endocrinol 109: 197–207, 1995

    Google Scholar 

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

    Google Scholar 

  25. Fuqua SAW, Chamness GC, McGuire WL: Estrogen receptor mutations in breast cancer. J Cell Biochem 51: 135–139, 1993

    Google Scholar 

  26. Daffada AA, Johnston SR, Smith IE, Detre S, King N, Dowsett M: Exon 5 deletion variant estrogen receptor messenger RNA expression in relation to tamoxifen resistance and progesterone receptor/pS2 status in human breast cancer. Cancer Res 55: 288–293, 1995

    Google Scholar 

  27. Villa E, Dugani A, Fantoni E, Camellini L, Buttafoco P, Grottola A, Pomper G, Santis MD, Ferrari A, Manenti F: Type of estrogen receptor determines response to antiestrogen therapy. Cancer Res 56: 3883–3885, 1996

    Google Scholar 

  28. Halachmi S, Marden E, Martin G, Mackay H, Abbondanza C, Brown M: Estrogen receptor associated proteins: possible mediators of hormone induced transcription. Science 264: 1455–1458, 1994

    Google Scholar 

  29. Yang NN, Venugopalan M, Hardikar S, Glasebrook A: Identification of an estrogen response element activated by metabolites of estradiol and raloxifene. Science 273: 1222–1225, 1996

    Google Scholar 

  30. Stierer M, Rosen H, Weber R, Hanak H, Auerbach L, Spona J, Tuchler H. Comparison of immunohistochemical and biochemical measurement of steriod receptors in primary breast cancer: Evaluation of discordant findings. Breast Cancer Res Treat 50: 125–134, 1998

    Google Scholar 

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Authors
  1. Aihua Huang
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  2. Etienne Leygue
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  3. Helmut Dotzlaw
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  4. Leigh C. Murphy
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  5. Peter H. Watson
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Huang, A., Leygue, E., Dotzlaw, H. et al. Influence of estrogen receptor variants on the determination of ER status in human breast cancer. Breast Cancer Res Treat 58, 217–223 (1999). https://doi.org/10.1023/A:1006375902601

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