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30.09.2019 | Brief Report

Optimized immunohistochemical detection of estrogen receptor beta using two validated monoclonal antibodies confirms its expression in normal and malignant breast tissues

verfasst von: John R. Hawse, Jodi M. Carter, Kirsten G. M. Aspros, Elizabeth S. Bruinsma, Justin W. Koepplin, Vivian Negron, Malayannan Subramaniam, James N. Ingle, Karen L. Rech, Matthew P. Goetz

Erschienen in: Breast Cancer Research and Treatment | Ausgabe 1/2020

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Abstract

Purpose

Significant controversy exists regarding the expression patterns of estrogen receptor beta (ERβ) in normal and diseased breast tissue. To address this issue, we have validated two ERβ antibodies, optimized the IHC protocols for both antibodies and now report the expression patterns of ERβ in normal and malignant breast tissues.

Methods

ERβ antibody specificity was determined using western blot and IHC analysis. ERβ protein expression patterns were assessed via IHC in normal breast tissue and invasive breast carcinoma. Further, we report the detailed protocol of the ERβ IHC assay developed in our CAP/CLIA certified laboratory to provide a standardized method for future studies.

Results

We have confirmed the specificity of two independent ERβ monoclonal antibodies, one that detects total (i.e., full length plus splice variants 2–5, which do not include the ligand binding domain) ERβ protein (PPZ0506) and one that detects only the full-length form, which includes the ligand binding domain, of ERβ (PPG5/10). Using these two antibodies, we demonstrate that ERβ is highly expressed in normal human breast tissue as well as in 20–30% of invasive breast cancers. Further, these two antibodies exhibited similar staining patterns across multiple different tissues and were highly concordant with regard to determining ERβ positivity in breast cancers.

Conclusions

ERβ protein was shown to be abundant in the majority of normal breast epithelial cells and is present in 20–30% of breast cancers. Use of these two antibodies, along with their standardized IHC protocols, provide a reference for future studies aimed at determining the utility of ERβ as a prognostic and/or predictive biomarker in various tissues of benign or malignant states.

Mosselman S, Polman J, Dijkema R (1996) ER beta: identification and characterization of a novel human estrogen receptor. FEBS Lett 392(1):49–53PubMed

Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA (1996) Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci USA 93(12):5925–5930PubMed

Haldosen LA, Zhao C, Dahlman-Wright K (2014) Estrogen receptor beta in breast cancer. Mol Cell Endocrinol 382(1):665–672PubMed

Nelson AW, Tilley WD, Neal DE, Carroll JS (2014) Estrogen receptor beta in prostate cancer: friend or foe? Endocr Relat Cancer 21(4):T219–T234PubMed

Guillette TC, Jackson TW, Belcher SM (2018) Duality of estrogen receptor beta action in cancer progression. Curr Opin Pharmacol 41:66–73PubMed

Warner M, Huang B, Gustafsson JA (2017) Estrogen receptor beta as a pharmaceutical target. Trends Pharmacol Sci 38(1):92–99PubMed

Wu X, Subramaniam M, Negron V, Cicek M, Reynolds C, Lingle WL, Goetz MP, Ingle JN, Spelsberg TC, Hawse JR (2012) Development, characterization, and applications of a novel estrogen receptor beta monoclonal antibody. J Cell Biochem 113(2):711–723PubMedPubMedCentral

Nelson AW, Groen AJ, Miller JL, Warren AY, Holmes KA, Tarulli GA, Tilley WD, Katzenellenbogen BS, Hawse JR, Gnanapragasam VJ et al (2017) Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity. Mol Cell Endocrinol 440:138–150PubMedPubMedCentral

Andersson S, Sundberg M, Pristovsek N, Ibrahim A, Jonsson P, Katona B, Clausson CM, Zieba A, Ramstrom M, Soderberg O et al (2017) Insufficient antibody validation challenges oestrogen receptor beta research. Nat Commun 8:15840PubMedPubMedCentral

10.

Lewandowski S, Kalita K, Kaczmarek L (2002) Estrogen receptor beta Potential functional significance of a variety of mRNA isoforms. FEBS Lett 524(1–3):1–5PubMed

11.

Moore JT, McKee DD, Slentz-Kesler K, Moore LB, Jones SA, Horne EL, Su JL, Kliewer SA, Lehmann JM, Willson TM (1998) Cloning and characterization of human estrogen receptor beta isoforms. Biochem Biophys Res Commun 247(1):75–78PubMed

12.

Poola I, Abraham J, Baldwin K, Saunders A, Bhatnagar R (2005) Estrogen receptors beta4 and beta5 are full length functionally distinct ERbeta isoforms: cloning from human ovary and functional characterization. Endocrine 27(3):227–238PubMed

13.

Weitsman GE, Skliris G, Ung K, Peng B, Younes M, Watson PH, Murphy LC (2006) Assessment of multiple different estrogen receptor-beta antibodies for their ability to immunoprecipitate under chromatin immunoprecipitation conditions. Breast Cancer Res Treat 100(1):23–31PubMed

14.

Wimberly H, Han G, Pinnaduwage D, Murphy LC, Yang XR, Andrulis IL, Sherman M, Figueroa J, Rimm DL (2014) ERbeta splice variant expression in four large cohorts of human breast cancer patient tumors. Breast Cancer Res Treat 146(3):657–667PubMedPubMedCentral

15.

Monroe DG, Getz BJ, Johnsen SA, Riggs BL, Khosla S, Spelsberg TC (2003) Estrogen receptor isoform-specific regulation of endogenous gene expression in human osteoblastic cell lines expressing either ERalpha or ERbeta. J Cell Biochem 90(2):315–326PubMed

16.

Reese JM, Suman VJ, Subramaniam M, Wu X, Negron V, Gingery A, Pitel KS, Shah SS, Cunliffe HE, McCullough AE et al (2014) ERbeta1: characterization, prognosis, and evaluation of treatment strategies in ERalpha-positive and -negative breast cancer. BMC Cancer 14:749PubMedPubMedCentral

17.

Secreto FJ, Monroe DG, Dutta S, Ingle JN, Spelsberg TC (2007) Estrogen receptor alpha/beta isoforms, but not betacx, modulate unique patterns of gene expression and cell proliferation in Hs578T cells. J Cell Biochem 101(5):1125–1147PubMed

18.

Leon-Ferre RA, Polley MY, Liu H, Gilbert JA, Cafourek V, Hillman DW, Elkhanany A, Akinhanmi M, Lilyquist J, Thomas A et al (2018) Impact of histopathology, tumor-infiltrating lymphocytes, and adjuvant chemotherapy on prognosis of triple-negative breast cancer. Breast Cancer Res Treat 167(1):89–99PubMed

19.

Jarvinen TA, Pelto-Huikko M, Holli K, Isola J (2000) Estrogen receptor beta is coexpressed with ERalpha and PR and associated with nodal status, grade, and proliferation rate in breast cancer. Am J Pathol 156(1):29–35PubMedPubMedCentral

20.

Skliris GP, Carder PJ, Lansdown MR, Speirs V (2001) Immunohistochemical detection of ERbeta in breast cancer: towards more detailed receptor profiling? Br J Cancer 84(8):1095–1098PubMedPubMedCentral

21.

Skliris GP, Munot K, Bell SM, Carder PJ, Lane S, Horgan K, Lansdown MR, Parkes AT, Hanby AM, Markham AF et al (2003) Reduced expression of oestrogen receptor beta in invasive breast cancer and its re-expression using DNA methyl transferase inhibitors in a cell line model. J Pathol 201(2):213–220PubMed

22.

Roger P, Sahla ME, Makela S, Gustafsson JA, Baldet P, Rochefort H (2001) Decreased expression of estrogen receptor beta protein in proliferative preinvasive mammary tumors. Cancer Res 61(6):2537–2541PubMed

23.

Shaaban AM, O’Neill PA, Davies MP, Sibson R, West CR, Smith PH, Foster CS (2003) Declining estrogen receptor-beta expression defines malignant progression of human breast neoplasia. Am J Surg Pathol 27(12):1502–1512PubMed

24.

Iwao K, Miyoshi Y, Egawa C, Ikeda N, Noguchi S (2000) Quantitative analysis of estrogen receptor-beta mRNA and its variants in human breast cancers. Int J Cancer 88(5):733–736PubMed

25.

Leygue E, Dotzlaw H, Watson PH, Murphy LC (1998) Altered estrogen receptor alpha and beta messenger RNA expression during human breast tumorigenesis. Cancer Res 58(15):3197–3201PubMed

26.

Leygue E, Dotzlaw H, Watson PH, Murphy LC (1999) Expression of estrogen receptor beta1, beta2, and beta5 messenger RNAs in human breast tissue. Cancer Res 59(6):1175–1179PubMed

27.

Zhao C, Lam EW, Sunters A, Enmark E, De Bella MT, Coombes RC, Gustafsson JA, Dahlman-Wright K (2003) Expression of estrogen receptor beta isoforms in normal breast epithelial cells and breast cancer: regulation by methylation. Oncogene 22(48):7600–7606PubMed

28.

Bardin A, Boulle N, Lazennec G, Vignon F, Pujol P (2004) Loss of ERbeta expression as a common step in estrogen-dependent tumor progression. Endocr Relat Cancer 11(3):537–551PubMedPubMedCentral

29.

Shaw JA, Udokang K, Mosquera JM, Chauhan H, Jones JL, Walker RA (2002) Oestrogen receptors alpha and beta differ in normal human breast and breast carcinomas. J Pathol 198(4):450–457PubMed

30.

Park BW, Kim KS, Heo MK, Ko SS, Hong SW, Yang WI, Kim JH, Kim GE, Lee KS (2003) Expression of estrogen receptor-beta in normal mammary and tumor tissues: is it protective in breast carcinogenesis? Breast Cancer Res Treat 80(1):79–85PubMed

31.

Miyoshi Y, Taguchi T, Gustafsson JA, Noguchi S (2001) Clinicopathological characteristics of estrogen receptor-beta-positive human breast cancers. Jpn J Cancer Res 92(10):1057–1061PubMedPubMedCentral

32.

Omoto Y, Inoue S, Ogawa S, Toyama T, Yamashita H, Muramatsu M, Kobayashi S, Iwase H (2001) Clinical value of the wild-type estrogen receptor beta expression in breast cancer. Cancer Lett 163(2):207–212PubMed

33.

Fuqua SA, Schiff R, Parra I, Moore JT, Mohsin SK, Osborne CK, Clark GM, Allred DC (2003) Estrogen receptor beta protein in human breast cancer: correlation with clinical tumor parameters. Cancer Res 63(10):2434–2439PubMedPubMedCentral

34.

Mann S, Laucirica R, Carlson N, Younes PS, Ali N, Younes A, Li Y, Younes M (2001) Estrogen receptor beta expression in invasive breast cancer. Hum Pathol 32(1):113–118PubMed

35.

Myers E, Fleming FJ, Crotty TB, Kelly G, McDermott EW, O’Higgins NJ, Hill AD, Young LS (2004) Inverse relationship between ER-beta and SRC-1 predicts outcome in endocrine-resistant breast cancer. Br J Cancer 91(9):1687–1693PubMedPubMedCentral

36.

Nakopoulou L, Lazaris AC, Panayotopoulou EG, Giannopoulou I, Givalos N, Markaki S, Keramopoulos A (2004) The favourable prognostic value of oestrogen receptor beta immunohistochemical expression in breast cancer. J Clin Pathol 57(5):523–528PubMedPubMedCentral

37.

Sugiura H, Toyama T, Hara Y, Zhang Z, Kobayashi S, Fujii Y, Iwase H, Yamashita H (2007) Expression of estrogen receptor beta wild-type and its variant ERbetacx/beta2 is correlated with better prognosis in breast cancer. Jpn J Clin Oncol 37(11):820–828PubMed

38.

Miller WR, Anderson TJ, Dixon JM, Saunders PT (2006) Oestrogen receptor beta and neoadjuvant therapy with tamoxifen: prediction of response and effects of treatment. Br J Cancer 94(9):1333–1338PubMedPubMedCentral

39.

Saunders PT, Millar MR, Williams K, Macpherson S, Bayne C, O’Sullivan C, Anderson TJ, Groome NP, Miller WR (2002) Expression of oestrogen receptor beta (ERbeta1) protein in human breast cancer biopsies. Br J Cancer 86(2):250–256PubMedPubMedCentral

40.

O’Neill PA, Davies MP, Shaaban AM, Innes H, Torevell A, Sibson DR, Foster CS (2004) Wild-type oestrogen receptor beta (ERbeta1) mRNA and protein expression in Tamoxifen-treated post-menopausal breast cancers. Br J Cancer 91(9):1694–1702PubMedPubMedCentral

41.

Skliris GP, Leygue E, Curtis-Snell L, Watson PH, Murphy LC (2006) Expression of oestrogen receptor-beta in oestrogen receptor-alpha negative human breast tumours. Br J Cancer 95(5):616–626PubMedPubMedCentral

42.

Jensen EV, Cheng G, Palmieri C, Saji S, Makela S, Van Noorden S, Wahlstrom T, Warner M, Coombes RC, Gustafsson JA (2001) Estrogen receptors and proliferation markers in primary and recurrent breast cancer. Proc Natl Acad Sci USA 98(26):15197–15202PubMed

43.

Poola I, Fuqua SA, De Witty RL, Abraham J, Marshallack JJ, Liu A (2005) Estrogen receptor alpha-negative breast cancer tissues express significant levels of estrogen-independent transcription factors, ERbeta1 and ERbeta5: potential molecular targets for chemoprevention. Clin Cancer Res 11(20):7579–7585PubMed

44.

Umekita Y, Souda M, Ohi Y, Sagara Y, Rai Y, Takahama T, Yoshida H (2006) Expression of wild-type estrogen receptor beta protein in human breast cancer: specific correlation with HER2/neu overexpression. Pathol Int 56(8):423–427PubMed

45.

Wang J, Zhang C, Chen K, Tang H, Tang J, Song C, Xie X (2015) ERbeta1 inversely correlates with PTEN/PI3K/AKT pathway and predicts a favorable prognosis in triple-negative breast cancer. Breast Cancer Res Treat 152(2):255–269PubMed

46.

Shanle EK, Zhao Z, Hawse J, Wisinski K, Keles S, Yuan M, Xu W (2013) Research resource: global identification of estrogen receptor beta target genes in triple negative breast cancer cells. Mol Endocrinol 27(10):1762–1775PubMedPubMedCentral

47.

Lazennec G, Bresson D, Lucas A, Chauveau C, Vignon F (2001) ER beta inhibits proliferation and invasion of breast cancer cells. Endocrinology 142(9):4120–4130PubMedPubMedCentral

48.

Thomas C, Rajapaksa G, Nikolos F, Hao R, Katchy A, McCollum CW, Bondesson M, Quinlan P, Thompson A, Krishnamurthy S et al (2012) ERbeta1 represses basal breast cancer epithelial to mesenchymal transition by destabilizing EGFR. Breast Cancer Res 14(6):R148PubMedPubMedCentral

49.

Wu X, Subramaniam M, Grygo SB, Sun Z, Negron V, Lingle WL, Goetz MP, Ingle JN, Spelsberg TC, Hawse JR (2011) Estrogen receptor-beta sensitizes breast cancer cells to the anti-estrogenic actions of endoxifen. Breast Cancer Res 13(2):27

50.

Anestis A, Sarantis P, Theocharis S, Zoi I, Tryfonopoulos D, Korogiannos A, Koumarianou A, Xingi E, Thomaidou D, Kontos M et al (2019) Estrogen receptor beta increases sensitivity to enzalutamide in androgen receptor-positive triple-negative breast cancer. J Cancer Res Clin Oncol 145(5):1221–1233PubMed

51.

Reese JM, Bruinsma ES, Monroe DG, Negron V, Suman VJ, Ingle JN, Goetz MP, Hawse JR (2017) ERβ inhibits cyclin dependent kinases 1 and 7 in triple negative breast cancer. Oncotarget 8(57):96506–96521PubMedPubMedCentral

52.

Reese JM, Bruinsma ES, Nelson AW, Chernukhin I, Carroll JS, Li Y, Subramaniam M, Suman VJ, Negron V, Monroe DG et al (2018) ERbeta-mediated induction of cystatins results in suppression of TGFbeta signaling and inhibition of triple-negative breast cancer metastasis. Proc Natl Acad Sci USA 111:E9580–E9589

53.

Bado I, Pham E, Soibam B, Nikolos F, Gustafsson JA, Thomas C (2018) ERbeta alters the chemosensitivity of luminal breast cancer cells by regulating p53 function. Oncotarget 9(32):22509–22522PubMedPubMedCentral

54.

Zhao L, Huang S, Mei S, Yang Z, Xu L, Zhou N, Yang Q, Shen Q, Wang W, Le X et al (2018) Pharmacological activation of estrogen receptor beta augments innate immunity to suppress cancer metastasis. Proc Natl Acad Sci USA 115(16):E3673–E3681PubMed

55.

Bado I, Nikolos F, Rajapaksa G, Wu W, Castaneda J, Krishnamurthy S, Webb P, Gustafsson JA, Thomas C (2017) Somatic loss of estrogen receptor beta and p53 synergize to induce breast tumorigenesis. Breast Cancer Res 19(1):79PubMedPubMedCentral

56.

Bado I, Nikolos F, Rajapaksa G, Gustafsson JA, Thomas C (2016) ERbeta decreases the invasiveness of triple-negative breast cancer cells by regulating mutant p53 oncogenic function. Oncotarget 7(12):13599–13611PubMedPubMedCentral

57.

Huang B, Warner M, Gustafsson JA (2015) Estrogen receptors in breast carcinogenesis and endocrine therapy. Mol Cell Endocrinol 418(Pt 3):240–244PubMed

58.

Esslimani-Sahla M, Simony-Lafontaine J, Kramar A, Lavaill R, Mollevi C, Warner M, Gustafsson JA, Rochefort H (2004) Estrogen receptor beta (ER beta) level but not its ER beta cx variant helps to predict tamoxifen resistance in breast cancer. Clin Cancer Res 10(17):5769–5776PubMed

59.

Fleming FJ, Hill AD, McDermott EW, O’Higgins NJ, Young LS (2004) Differential recruitment of coregulator proteins steroid receptor coactivator-1 and silencing mediator for retinoid and thyroid receptors to the estrogen receptor-estrogen response element by beta-estradiol and 4-hydroxytamoxifen in human breast cancer. J Clin Endocrinol Metab 89(1):375–383PubMed

60.

Hopp TA, Weiss HL, Parra IS, Cui Y, Osborne CK, Fuqua SA (2004) Low levels of estrogen receptor beta protein predict resistance to tamoxifen therapy in breast cancer. Clin Cancer Res 10(22):7490–7499PubMed

61.

Iwase H, Zhang Z, Omoto Y, Sugiura H, Yamashita H, Toyama T, Iwata H, Kobayashi S (2003) Clinical significance of the expression of estrogen receptors alpha and beta for endocrine therapy of breast cancer. Cancer Chemother Pharmacol 52(Suppl 1):S34–S38PubMed

62.

Murphy LC, Leygue E, Niu Y, Snell L, Ho SM, Watson PH (2002) Relationship of coregulator and oestrogen receptor isoform expression to de novo tamoxifen resistance in human breast cancer. Br J Cancer 87(12):1411–1416PubMedPubMedCentral

63.

Speirs V, Parkes AT, Kerin MJ, Walton DS, Carleton PJ, Fox JN, Atkin SL (1999) Coexpression of estrogen receptor alpha and beta: poor prognostic factors in human breast cancer? Cancer Res 59(3):525–528PubMed

64.

Speirs V, Malone C, Walton DS, Kerin MJ, Atkin SL (1999) Increased expression of estrogen receptor beta mRNA in tamoxifen-resistant breast cancer patients. Cancer Res 59(21):5421–5424PubMed

65.

Honma N, Horii R, Iwase T, Saji S, Younes M, Takubo K, Matsuura M, Ito Y, Akiyama F, Sakamoto G (2008) Clinical importance of estrogen receptor-beta evaluation in breast cancer patients treated with adjuvant tamoxifen therapy. J Clin Oncol 26(22):3727–3734PubMed

66.

Shaaban AM, Green AR, Karthik S, Alizadeh Y, Hughes TA, Harkins L, Ellis IO, Robertson JF, Paish EC, Saunders PT et al (2008) Nuclear and cytoplasmic expression of ERbeta1, ERbeta2, and ERbeta5 identifies distinct prognostic outcome for breast cancer patients. Clin Cancer Res 14(16):5228–5235PubMed

67.

Novelli F, Milella M, Melucci E, Di Benedetto A, Sperduti I, Perrone-Donnorso R, Perracchio L, Venturo I, Nistico C, Fabi A et al (2008) A divergent role for estrogen receptor-beta in node-positive and node-negative breast cancer classified according to molecular subtypes: an observational prospective study. Breast Cancer Res 10(5):R74PubMedPubMedCentral

68.

Motomura K, Ishitobi M, Komoike Y, Koyama H, Nagase H, Inaji H, Noguchi S (2010) Expression of estrogen receptor beta and phosphorylation of estrogen receptor alpha serine 167 correlate with progression-free survival in patients with metastatic breast cancer treated with aromatase inhibitors. Oncology 79(1–2):55–61PubMed

Titel: Optimized immunohistochemical detection of estrogen receptor beta using two validated monoclonal antibodies confirms its expression in normal and malignant breast tissues
verfasst von: John R. Hawse
Jodi M. Carter
Kirsten G. M. Aspros
Elizabeth S. Bruinsma
Justin W. Koepplin
Vivian Negron
Malayannan Subramaniam
James N. Ingle
Karen L. Rech
Matthew P. Goetz
Publikationsdatum: 30.09.2019
Verlag: Springer US
Erschienen in: Breast Cancer Research and Treatment / Ausgabe 1/2020
Print ISSN: 0167-6806
Elektronische ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-019-05441-3

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25.04.2024 | Nierenkarzinom | Nachrichten

Springer Medizin

Optimized immunohistochemical detection of estrogen receptor beta using two validated monoclonal antibodies confirms its expression in normal and malignant breast tissues

Abstract

Purpose

Methods

Results

Conclusions

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Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

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