Regulation of estrogen receptor β1 expression in breast cancer by epigenetic modification of the 5' regulatory region

Al-Nakhle,Hakeemah; Smith,Laura; Bell,Sandra  M.; Burns,Philip   A.; Cummings,Michele; Hanby,Andrew  M.; Lane,Sally; Parker,Marie   D.; Hughes,Thomas   A.; Speirs,Valerie

doi:10.3892/ijo.2013.2112

Regulation of estrogen receptor β1 expression in breast cancer by epigenetic modification of the 5' regulatory region

Authors:
- Hakeemah Al-Nakhle
- Laura Smith
- Sandra M. Bell
- Philip A. Burns
- Michele Cummings
- Andrew M. Hanby
- Sally Lane
- Marie D. Parker
- Thomas A. Hughes
- Valerie Speirs
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Affiliations: Leeds Institute of Cancer and Pathology, University of Leeds, Leeds LS9 7TF, UK, Leeds Institute of Biomedical and Clinical Sciences, University of Leeds, Leeds LS9 7TF, UK, St James's Institute of Oncology, St James's University Hospital, Leeds LS9 7TF, UK
Published online on: September 25, 2013 https://doi.org/10.3892/ijo.2013.2112
Pages: 2039-2045

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Abstract

ERβ1 is often down-regulated in breast cancer compared to normal breast but mechanisms surrounding this are unclear. We examined whether loss of heterozygosity (LOH) or methylation at ERβ promoters (0N, 0K) and/or untranslated exon 0N were involved in ERβ down-regulation in breast cancer tissues and cell lines and if treatment with the de-methylating agent 5-aza-deoxycytidine and/or the histone deacetylase inhibitor Trichostatin A could influence expression in vitro. We found no evidence of correlation between LOH at 14q22-24 (genomic locus containing ERβ/ESR2), and ERβ1 expression in primary breast cancers. A negative correlation between ERβ1 mRNA expression and methylation status was observed for promoter 0N in BT-20, MDA-MB-453 and T47D cells. Promoter 0K was consistently unmethylated. In primary breast tumours, methylation of the untranslated exon 0N, downstream of promoter 0N, but not of promoter 0N itself, correlated with down-regulation of ERβ. In MDA-MB-453 cells, treatment with 5-aza-deoxycytidine was sufficient to induce ERβ1 expression from the 0N promoter while in BT-20 both agents were required. Examination of various sites on ESR2 highlighted epigenetic but not genetic regulation of ERβ1. In particular methylation adjacent to promoter 0N was a key regulatory event for ERβ1 silencing. A combination of de-methylating agents and histone deacetylase inhibitors fully restored ERβ1 expression which may offer a novel therapeutic angle for breast cancer management.

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1.	Moore J, McKee D, Slentz-Kesler K, Moore LB, Jones S, Horne E, Su J, Kliewer S, Lehmann J and Willson T: Cloning and characterization of human estrogen receptor β isoforms. Biochem Biophys Res Commun. 247:75–78. 1998.
2.	Leung Y, Mak P, Hassan S and Ho S: Estrogen receptor (ER)-β isoforms: a key to understanding ER-β signaling. Proc Natl Acad Sci USA. 103:13162–13167. 1996.
3.	Speirs V, Skliris GP, Burdall SE and Carder PJ: Distinct expression patterns of ERα and ERβ in normal human mammary gland. J Clin Pathol. 55:371–374. 2002.
4.	Skliris GP, Munot K, Bell SM, Carder PJ, Lane S, Horgan K, Lansdown MR, Parkes AT, Hanby AM, Markham AF and Speirs V: Reduced expression of oestrogen receptor β in invasive breast cancer and its re-expression using DNA methyl transferase inhibitors in a cell line model. J Pathol. 201:213–220. 2003.
5.	Matthews J and Gustafsson J: Estrogen signaling: a subtle balance between ERα and ERβ. Mol Interv. 3:281–292. 2003.PubMed/NCBI
6.	Bardin A, Boulle N, Lazennec G, Vignon F and Pujol P: Loss of ERβ expression as a common step in estrogen-dependent tumor progression. Endocr Relat Cancer. 11:537–551. 2004.
7.	Enmark E, Pelto-Huikko M, Grandien K, Lagercrantz S, Lagercrantz J, Fried G, Nordenskjold M and Gustafsson JA: Human estrogen receptor beta: gene structure, chromosomal localization, and expression pattern. J Clin Endocrinol Metab. 82:4258–4265. 1997.PubMed/NCBI
8.	Suzuki F, Akahira J, Miura I, Suzuki T, Ito K, Hayashi H, Sasano H and Yaegashi N: Loss of estrogen receptor β isoform expression and its correlation with aberrant DNA methylation of the 5′-untranslated region in human epithelial ovarian carcinoma. Cancer Sci. 99:2365–2372. 2008.
9.	Yap OW, Bhat G, Liu L and Tollefsbol TO: Epigenetic modifications of the estrogen receptor β gene in epithelial ovarian cancer cells. Anticancer Res. 29:139–144. 2009.
10.	Zhao C, Lam EW, Sunters A, Enmark E, De Bella MT, Coombes RC, Gustafsson JA and Dahlman-Wright K: Expression of estrogen receptor β isoforms in normal breast epithelial cells and breast cancer: regulation by methylation. Oncogene. 22:7600–7606. 2003.
11.	Zhu X, Leav I, Leung YK, Wu M, Liu Q, Gao Y, McNeal JE and Ho SM: Dynamic regulation of estrogen receptor-β expression by DNA methylation during prostate cancer development and metastasis. Am J Pathol. 164:2003–2012. 2004.
12.	Rody A, Holtrich U, Solbach C, Kourtis K, von Minckwitz G, Engels K, Kissler S, Gätje R, Karn T and Kaufmann M: Methylation of estrogen receptor β promoter correlates with loss of ER-β expression in mammary carcinoma and is an early indication marker in premalignant lesions. Endocr Relat Cancer. 12:903–916. 2005.
13.	Smith L, Brannan RA, Hanby AM, Shaaban AM, Verghese ET, Peter MB, Pollock S, Satheesha S, Szynkiewicz M, Speirs V and Hughes TA: Differential regulation of oestrogen receptor β isoforms by 5′ untranslated regions in cancer. J Cell Mol Med. 14:2172–2184. 2010.
14.	Zambrano P, Segura-Pacheco B, Perez-Cardenas E, Cetina L, Revilla-Vazquez A, Taja-Chayeb L, Chavez-Blanco A, Angeles E, Cabrera G, Sandoval K, Trejo-Becerril C, Chanona-Vilchis J and Duenas-González A: A phase I study of hydralazine to demethylate and reactivate the expression of tumor suppressor genes. BMC Cancer. 5:442005. View Article : Google Scholar : PubMed/NCBI
15.	Walton TJ, Li G, Seth R, McArdle SE, Bishop MC and Rees RC: DNA demethylation and histone deacetylation inhibition co-operate to re-express estrogen receptor β and induce apoptosis in prostate cancer cell-lines. Prostate. 68:210–222. 2008.PubMed/NCBI
16.	Shaaban AM, Green AR, Karthik S, Alizadeh Y, Hughes TA, Harkins L, Ellis IO, Robertson JF, Paish EC, Saunders PT, Groome NP and Speirs V: Nuclear and cytoplasmic expression of ERβ1, ERβ2, and ERβ5 identifies distinct prognostic outcome for breast cancer patients. Clin Cancer Res. 14:5228–5235. 2008.
17.	Cummings M, Iremonger J, Green CA, Shaaban AM and Speirs V: Gene expression of ERβ isoforms in laser microdissected human breast cancers: implications for gene expression analyses. Cell Oncol. 31:467–473. 2009.
18.	Cawkwell L, Bell SM, Lewis FA, Dixon MF, Taylor GR and Quirke P: Rapid detection of allele loss in colorectal tumours using microsatellites and fluorescent DNA technology. Br J Cancer. 67:1262–1267. 1993. View Article : Google Scholar : PubMed/NCBI
19.	Cottrell SE, Distler J, Goodman NS, Mooney SH, Kluth A, Olek A, Schwope I, Tetzner R, Ziebarth H and Berlin K: A real-time PCR assay for DNA-methylation using methylation-specific blockers. Nucleic Acids Res. 32:e102004. View Article : Google Scholar : PubMed/NCBI
20.	Smith L, Coleman LJ, Cummings M, Satheesha S, Shaw SO, Speirs V and Hughes TA: Expression of estrogen receptor β isoforms is regulated by transcriptional and post-transcriptional mechanisms. Biochem J. 429:283–290. 2010.
21.	Herman JG, Graff JR, Myohanen S, Nelkin BD and Baylin SB: Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 93:9821–9826. 1996. View Article : Google Scholar : PubMed/NCBI
22.	Xue Q, Lin Z, Cheng YH, Huang CC, Marsh E, Yin P, Milad MP, Confino E, Reierstad S, Innes J and Bulun SE: Promoter methylation regulates estrogen receptor 2 in human endometrium and endometriosis. Biol Reprod. 77:681–687. 2007. View Article : Google Scholar : PubMed/NCBI
23.	Mirza S, Sharma G, Parshad R, Srivastava A, Gupta SD and Ralhan R: Clinical significance of promoter hypermethylation of ERβ and RARβ2 in tumor and serum DNA in Indian breast cancer patients. Ann Surg Oncol. 19:3107–3115. 2012.
24.	Nojima D, Li LC, Dharia A, Perinchery G, Ribeiro-Filho L, Yen TS and Dahiya R: CpG hypermethylation of the promoter region inactivates the estrogen receptor-β gene in patients with prostate carcinoma. Cancer. 92:2076–2083. 2001.PubMed/NCBI
25.	Zhai RL, Wang GB, Cai KL, Tao KX, Xu F, Zhang WL and Zang ZY: Transcriptional inactivation of ERβ gene is mediated by the induction of promoter hypermethylation in a rat colonic epithelial cell model. J Surg Res. 155:306–310. 2009.
26.	Hamilton-Burke W, Coleman LJ, Cummings M, Green CA, Holliday DL, Horgan K, Maraqa L, Peter MB, Pollock S, Shaaban AM, Smith L and Speirs V: Phosphorylation of estrogen receptor β at serine 105 is associated with good prognosis in breast cancer. Am J Pathol. 177:1079–1086. 2010.
27.	Al-Nakhle H, Burns PA, Cummings M, Hanby AM, Hughes TA, Satheesha S, Shaaban AM, Smith L and Speirs V: Estrogen receptor β1 expression is regulated by miR-92 in breast cancer. Cancer Res. 70:4778–4784. 2010.
28.	Bernstein I, Byun HM, Mohrbacher A, Douer D, Gorospe G III, Hergesheimer J, Groshen S, O’Connell C and Yang AS: A phase I biological study of azacitidine (Vidaza™) to determine the optimal dose to inhibit DNA methylation. Epigenetics. 5:750–757. 2010.
29.	Yeo W, Chung HC, Chan SL, Wang LZ, Lim R, Picus J, Boyer M, Mo FK, Koh J, Rha SY, Hui EP and Jeung HC: Epigenetic therapy using belinostat for patients with unresectable hepatocellular carcinoma: a multicenter phase I/II study with biomarker and pharmacokinetic analysis of tumors from patients in the Mayo Phase II Consortium and the Cancer Therapeutics Research Group. J Clin Oncol. 30:3361–3367. 2012.