Abstract
Human estrogen receptors alpha and beta are crucially involved in the regulation of mammary growth and development. Normal breast tissues display a relative higher expression of ER beta than ER alpha, which drastically changes during breast tumorogenesis. Thus, it is reasonable to suggest that a dysregulation of the two estrogen receptor subtypes may induce breast cancer development. However, the molecular mechanisms underlying the potential opposing roles played by the two estrogen receptors on tumor cell growth remain to be elucidated. In the present study, we have demonstrated that ER beta overexpression in breast cancer cells decreases cell proliferation and down-regulates ER alpha mRNA and protein content, along with a concomitant repression of estrogen-regulated genes. Transient transfection experiments, using a vector containing the human ER alpha promoter region, showed that elevated levels of ER beta down-regulated basal ER alpha promoter activity. Furthermore, site-directed mutagenesis and deletion analysis revealed that the proximal GC-rich motifs at −223 and −214 are critical for the ER beta-induced ER alpha down-regulation in breast cancer cells. This occurred through ER beta-Sp1 protein–protein interactions within the ER alpha promoter region and the recruitment of a corepressor complex containing the nuclear receptor corepressor NCoR, accompanied by hypoacetylation of histone H4 and displacement of RNA-polymerase II. Silencing of NCoR gene expression by RNA interference reversed the down-regulatory effects of ER beta on ER alpha gene expression and cell proliferation. Our results provide evidence for a novel mechanism by which overexpression of ER beta through NCoR is able to down regulate ER alpha gene expression, thus blocking ER alpha’s driving role on breast cancer cell growth.
Similar content being viewed by others
References
Hall JM, Couse JF, Korach KS (2001) The multifaceted mechanisms of estradiol and estrogen receptor signaling. J Biol Chem 276:36869–36872
Heldring N, Pike A, Andersson S et al (2007) Estrogen receptors: how do they signal and what are their targets. Physiol Rev 87:905–931
Osborne CK, Schiff R, Fuqua SA et al (2001) Estrogen receptor: current understanding of its activation and modulation. Clin Cancer Res 7:4338s–4342s (discussion 4411s–4412s)
Barone I, Brusco L, Fuqua SA (2010) Estrogen receptor mutations and changes in downstream gene expression and signaling. Clin Cancer Res 16:2702–2708 Review
Cato AC, Nestl A, Mink S (2002) Rapid actions of steroid receptors in cellular signaling pathways. Science STKE 138:re9
Nemere I, Pietras RJ, Blackmore PF (2003) Membrane receptors for steroid hormones: signal transduction and physiological significance. J Cell Biochem 88:438–445
Ordonez-Moran P, Munoz A (2009) Nuclear receptors: genomic and nongenomic effects converge. Cell Cycle 8:1675–1680
Paech K, Webb P, Kuiper GG et al (1997) Differential ligand activation of estrogen receptors ERα and ERβ at AP1 sites. Science 277:1508–1510
Webb P, Nguyen P, Valentine C et al (1999) The estrogen receptor enhances AP-1 activity by two distinct mechanisms with different requirements for receptor transactivation functions. Mol Endocrinol 13:1672–1685
Kushner PJ, Agard DA, Greene GL et al (2000) Estrogen receptor pathways to AP-1. J Steroid Biochem Mol Biol 74:311–317
Salvatori L, Pallante P, Ravenna L et al (2003) Oestrogens and selective oestrogen receptor (ER) modulators regulate EGF receptor gene expression through human ER α and β subtypes via an Sp1 site. Oncogene 22:4875–4881
Safe S, Kim K (2004) Nuclear receptor-mediated transactivation through interaction with Sp proteins. Nucleic Acid Res Mol Biol 77:1–36
Panno ML, Mauro L, Marsico S et al (2006) Evidence that mouse IRS-1 belongs to the family gene which promoter is activated by ERα through its interaction with Sp-1. J Mol Endocrinol 36:91–105
Catalano S, Rizza P, Gu G et al (2007) Fas ligand expression in TM4 Sertoli cells is enhanced by estradiol in situ production. J Cell Physiol 211:448–456
Mauro L, Catalano S, Bossi G et al (2007) Evidences that leptin upregulates E-cadherin expression in breast cancer effects on tumor growth and progression. Cancer Res 67:3412–3421
Higgins KJ, Shengxi L, Abdelrahim M et al (2008) Vascular endothelial growth factor receptor-2 expression is down-regulated by 17 beta estradiol in MCF-7 breast cancer cells by estrogen receptor alpha/Sp-1 proteins. Mol Endocrinol 22:388–402
Varlakhanova N, Snyder C, Jose S et al (2010) Estrogen receptors recruit SMRT and N-CoR corepressors through newly recognized contacts between the corepressor N terminus and the receptor DNA binding domain. Mol Cell Biol 30:1434–1445
Allegra JC, Lippman ME, Green L et al (1979) Estrogen receptor values in patients with benign breast disease. Cancer 44:228–231
Petersen OW, Hoyer PE, van Deurs B (1987) Frequency and distribution of estrogen receptor-positive cells in normal, nonlactating human breast tissue. Cancer Res 47:5748–5751
Ricketts D, Turnbull L, Ryall G et al (1991) Estrogen and progesterone receptors in the normal female breast. Cancer Res 51:1817–1822
Yager JD, Davidson NE (2006) Estrogen carcinogenesis in breast cancer. N Engl J Med 354:270–282
Zhao C, Dahlman-Wright K et al (2008) Estrogen receptor beta: an overview and update. Nucl Recept Signal 6:e003
Skliris GP, Munot K, Bell SM et al (2003) Reduced expression of estrogen receptor beta in invasive breast cancer and its re-expression using DNA methyl transferase inhibitors in a cell line model. J Pathol 201:213–220
Hopp TA, Weiss HL, Parra IS et al (2004) Low levels of estrogen receptor beta protein predict resistance to tamoxifen therapy in breast cancer. Clin Cancer Res 10:7490–7499
Paruthiyil S, Parmar H, Kerekatte V et al (2004) Estrogen receptor β inhibits human breast cancer cell proliferation and tumor formation by causing a G2 Cell Cycle Arrest. Cancer Res 64:423–428
Chang EC, Frasor J, Komm B et al (2006) Impact of estrogen receptor β on gene networks regulated by estrogen receptor α in breast cancer cells. Endocrinology 147:4831–4842
Williams C, Edvardsson K, Lewandowski SA et al (2008) A genome-wide study of repressive effects of estrogen receptor beta on estrogen receptor alpha signaling in breast cancer cells. Oncogene 27:1019–1032
Ström A, Hartman J, Foster JS et al (2004) Estrogen receptor β inhibits 17β-estradiol-stimulated proliferation of the breast cancer cell line T47D. Proc Natl Acad Sci 101:1566–1571
Hartman J, Lindberg K, Morani A et al (2006) Estrogen receptor beta inhibits angiogenesis and growth of T47D breast cancer xenografts. Cancer Res 66:11207–11213
Saji S, Omoto Y, Shimizu C et al (2002) Expression of estrogen receptor (ER) (beta)cx protein in ER(alpha)-positive breast cancer: specific correlation with progesterone receptor. Cancer Res 62:4849–4853
Buteau-Lozano H, Ancelin M, Lardeux B et al (2002) Transcriptional regulation of vascular endothelial growth factor by estradiol and tamoxifen in breast cancer cells: a complex interplay between estrogen receptors α and β. Cancer Res 62:4977–4984
Omoto Y, Eguchi H, Yamamoto-Yamaguchi Y et al (2003) Estrogen receptor (ER) β1 and ERβcx/β ERα function differently in breast cancer cell line MCF7. Oncogene 22:5011–5020
Pettersson K, Delaunay F, Gustafsson JA (2000) Estrogen receptor β acts as a dominant regulator of estrogen signaling. Oncogene 19:4970–4978
Nilsson S, Makela S, Treuter E et al (2001) Mechanisms of estrogen action. Physiol Rev 81:1535–1565
Liu MM, Albanese C, Anderson CM et al (2002) Opposing action of estrogen receptors alpha and beta on cyclin D1 gene expression. J Biol Chem 277:24353–24360
Acconcia F, Totta P, Ogawa S et al (2005) Survival versus apoptotic 17beta-estradiol effect: role of ER alpha and ER beta activated non-genomic signaling. J Cell Physiol 203:193–201
deGraffenried LA, Hopp TA, Valente AJ et al (2004) Regulation of the estrogen receptor α minimal promoter by Sp1, USF-1 and ERα. Breast Cancer Res and Treat 85:111–120
Catalano S, Malivindi R, Giordano C et al (2010) Farnesoid X Receptor through its binding to Steroidogenic Factor 1 responsive element inhibits aromatase expression in tumor Leydig cells. J Biol Chem 285:5581–5593
Andrews NC, Faller DV (1991) A rapid microperation technique foe extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res 19:2499
Barone I, Cui Y, Herynk MH et al (2009) Expression of the K303R estrogen receptor-alpha breast cancer mutation induces resistance to an aromatase inhibitor via addiction to the PI3 K/Akt kinase pathway. Cancer Res 69:4724–4732
Chen JD, Evans RM (1995) A transcriptional co-repressor that interacts with nuclear hormone receptors. Nature 377:454–457
Zamir I, Dawson J, Lavinsky RM et al (1997) Cloning and characterization of a corepressor and potential component of the nuclear hormone receptor repression complex. Proc Natl Acad Sci USA 94:14400–14405
Huynh KD, Fischle W, Verdin E et al (2000) BCoR, a novel corepressor involved in BCL-6 repression. Genes Dev 14:1810–1823
Lee JA, Suh DC, Kang JE et al (2005) Transcriptional activity of Sp1 is regulated by molecular interactions between the zinc finger DNA binding domain and the inhibitory domain with corepressors, and this interaction is modulated by MEK. J Biol Chem 280:28061–28071
Panno ML, Salerno M, Pezzi V et al (1996) Effect of estradiol and insulin on proliferative pattern and on estrogen and progesterone receptor contents in MCF-7 cells. J Cancer Res Clin Oncol 122:745–749
Lee AV, Weng C-N, Jackson JG et al (1997) Activation of estrogen receptor-mediated gene transcription by IGF-I in human breast cancer cells. J Endocrinol 152:39–47
Andò S, Panno ML, Salerno M et al (1998) Role of IRS-1 signaling in insulin-induced modulation of estrogen receptors in breast cancer cells. Biochem Biophys Res Commun 253:315–319
Lannigan DA (2003) Estrogen receptor phosphorylation. Steroids 68:1–9
Leygue E, Dotzlaw H, Watson PH et al (1998) Altered estrogen receptor α and β messenger RNA expression during human breast tumorigenesis. Cancer Res 58:3197–3201
Jarvinen TA, Pelto-Huikko M, Holli K et al (2000) Estrogen receptor β is coexpressed with ERα and PR and associated with nodal status, grade, and proliferation rate in breast cancer. Am J Pathol 156:29–35
Roger P, Sahla ME, Makela S et al (2001) Decreased expression of estrogen receptor β protein in proliferative preinvasive mammary tumors. Cancer Res 61:2537–2541
Fixemer T, Remberger K, Bonkhoff H (2003) Differential expression of the estrogen receptor beta in human prostate tissue, premalignant changes, and in primary, metastatic, and recurrent prostatic adenocarcinoma. Prostate 54:79–87
Foley EF, Jazaeri AA, Shupnik MA et al (2000) Selective loss of estrogen receptor β in malignant human colon. Cancer Res 60:245–248
Pujol P, Rey JM, Nirde P et al (1998) Differential expression of estrogen receptor-alpha and -beta messenger RNAs as a potential marker of ovarian carcinogenesis. Cancer Res 58:5367–5373
Behrens D, Gill JH, Fichtner I (2007) Loss of tumourigenicity of stably ERβ-transfected MCF-7 breast cancer cells. Mol Cell Endocrinol 274:19–29
Song K, Pan ZZ (2012) Estrogen receptor beta agonist diarylpropionitrile counteracts the estrogenic activity of estrogen receptor alpha agonist propylpyrazole-triol in the mammary gland of ovariectomized Sprague Dawley rats. J Steroid Biochem Mol Biol 130:26–35
Mann S, Laucirica R, Carlson N et al (2001) Estrogen receptor beta expression in invasive breast cancer. Hum Pathol 32:113–118
Murphy LC, Leygue E, Niu Y et al (2002) Relationship of coregulator and oestrogen receptor isoform expression to de novo tamoxifen resistance in human breast cancer. Br J Cancer 87:1411–1416
Esslimani-Sahla M, Simony-Lafontaine J, Kramar A et al (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 10:5769–5776
Saville B, Wormke M, Wang F et al (2000) Ligand-, cell-, and estrogen receptor subtype (alpha/beta)-dependent activation at GC-rich (Sp1) promoter elements. J Biol Chem 275:5379–5387
Porter W, Saville B, Hoivik D et al (1997) Functional synergy between the transcription factor Sp1 and the estrogen receptor. Mol Endocrinol 11:1569–1580
Duan R, Porter W, Safe S (1998) Estrogen-induced c-fos protooncogene expression in MCF-7 human breast cancer cells: role of estrogen receptor Sp1 complex formation. Endocrinology 139:1981–1990
Vyhlidal C, Samudio I, Kladde MP et al (2000) Transcriptional activation of transforming growth factor alpha by estradiol: requirement for both a GC-rich site and an estrogen response half-site. J Mol Endocrinol 24:329–338
Nagy L, Kao HY, Chakravarti D et al (1997) Nuclear receptor repression mediated by a complex containing SMRT, mSin3A, and histone deacetylase. Cell 89:373–380
Hu X, Lazar MA (1999) The CoRNR motif controls the recruitment of corepressors by nuclear hormone receptors. Nature 402:93–96
Webb P, Anderson CM, Valentine C et al (2000) The nuclear receptor corepressor (N-CoR) contains three isoleucine motifs (I/LXXII) that serve as receptor interaction domains (IDs). Mol Endocrinol 14:1976–1985
Jiang S, Meyer R, Kang K et al (2006) Scaffold attachment factor SAFB1 suppresses estrogen receptor α-mediated transcription in part via interaction with nuclear receptor corepressor. Mol Endocrinol 20:311–320
De Amicis F, Zupo S, Panno ML et al (2009) Progesterone receptor B recruits a repressor complex to a half-TRE site of the estrogen receptor α gene promoter. Mol Endocrinol 23:454–465
Ghisletti S, Huang W, Jepsen K et al (2009) Cooperative NCoR/SMRT interactions establish a corepressor-based strategy for integration of inflammatory and anti-inflammatory signaling pathways. Gene Dev 23:681–693
Acknowledgments
This study was supported by PRIN 2009 and AIRC grants 2011. Reintegration AIRC/Marie Curie International Fellowship in Cancer Research to IB. NIH/NCI R01 CA72038 to SAWF.
Conflict of interest
The authors declare no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
V. Bartella and P. Rizza have contributed equally to this study.
An erratum to this article is available at http://dx.doi.org/10.1007/s10549-016-3753-8.
Rights and permissions
About this article
Cite this article
Bartella, V., Rizza, P., Barone, I. et al. Estrogen receptor beta binds Sp1 and recruits a corepressor complex to the estrogen receptor alpha gene promoter. Breast Cancer Res Treat 134, 569–581 (2012). https://doi.org/10.1007/s10549-012-2090-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10549-012-2090-9