Expression of estrogen receptor β isoforms in normal breast epithelial cells and breast cancer: regulation by methylation

ABSTRACT Two novel estrogen receptor _β_ (ER_β_) mRNA isoforms that diverge in their 5′-untranslated regions, ER_β_ mRNA (0K-1) and ER_β_ mRNA (0N-1), have recently been identified. This

indicates that transcription of the human ER_β_ gene occurs from at least two different promoters, named promoter 0K and promoter 0N. The aim of this study was to investigate the expression

of ER_β_ isoforms in primary cultures of normal breast epithelial cells, a panel of breast cancer cell lines and in normal breast and breast cancer tissues; and to examine whether

methylation of the two ER_β_ promoters is involved in regulation of ER_β_ gene expression. Using quantitative real-time PCR techniques, we found that ER_β_ mRNA levels were significantly

lower in breast cancer cell lines than in primary cultures of normal breast epithelial cells. Bisulfite genomic sequencing analysis revealed that two promoters of the ER_β_ gene exhibit

distinct methylation patterns. Promoter 0N was unmethylated in normal breast epithelial cells, but extensively methylated in breast cancer cell lines. In contrast, promoter 0K was

unmethylated in both normal and malignant breast epithelial cells. We demonstrated a significant correlation between promoter 0N hypermethylation and loss of ER_β_ mRNA expression in breast

cancer cell lines. Treatment of breast cancer cells with demethylating agent effectively reactivated the expression of ER_β_ mRNA. The observations from the cell lines were also reflected in

primary breast cancer tumors. Thus, expression of ER_β_ mRNA in breast tumors was found to be inversely associated with the degree of methylation of promoter 0N. Our results suggest that a

decreased level of ER_β_ mRNA may be associated with breast tumorigenesis, and that DNA methylation is an important mechanism for ER_β_ gene silencing in breast cancer. Access through your

institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your institution Subscribe to this journal Receive 50 print

issues and online access $259.00 per year only $5.18 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to

local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT

BEING VIEWED BY OTHERS ABERRANT PROMOTER METHYLATION CONTRIBUTES TO _LRIG1_ SILENCING IN BASAL/TRIPLE-NEGATIVE BREAST CANCER Article Open access 19 April 2022 COMMON DNA METHYLATION DYNAMICS

IN ENDOMETRIOD ADENOCARCINOMA AND GLIOBLASTOMA SUGGEST UNIVERSAL EPIGENOMIC ALTERATIONS IN TUMORIGENESIS Article Open access 21 May 2021 MULTI-OMICS ANALYSES IDENTIFY _HSD17B4_

METHYLATION-SILENCING AS A PREDICTIVE AND RESPONSE MARKER OF HER2-POSITIVE BREAST CANCER TO HER2-DIRECTED THERAPY Article Open access 23 September 2020 REFERENCES * Bieche I, Parfait B,

Laurendeau I, Girault I, Vidaud M and Lidereau R . (2001). _Oncogene_, 20, 8109–8115. * Dotzlaw H, Leygue E, Watson PH and Murphy LC . (1999). _Cancer Res._, 59, 529–532. * Flouriot G,

Griffin C, Kenealy M, Sonntag-Buck V and Gannon F . (1998). _Mol. Endocrinol._, 12, 1939–1954. * Fuqua SA, Schiff R, Parra I, Friedrichs WE, Su JL, McKee DD, Slentz-Kesler K, Moore LB,

Willson TM and Moore JT . (1999). _Cancer Res._, 59, 5425–5428. * Gardiner-Garden M and Frommer M . (1987). _J. Mol. Biol._, 196, 261–282. * Garinis GA, Patrinos GP, Spanakis NE and Menounos

PG . (2002). _Hum. Genet._, 111, 115–127. * Gomm JJ, Browne PJ, Coope RC, Liu QY, Buluwela L and Coombes RC . (1995). _Anal. Biochem._, 226, 91–99. * Gustafsson JA and Warner M . (2000).

_J. Steroid Biochem. Mol. Biol._, 74, 245–248. * Herman JG, Graff JR, Myohanen S, Nelkin BD and Baylin SB . (1996). _Proc. Natl. Acad. Sci. USA_, 93, 9821–9826. * Hirata S, Shoda T, Kato J

and Hoshi K . (2001). _J. Steroid Biochem. Mol. Biol._, 78, 33–40. * Iwao K, Miyoshi Y, Egawa C, Ikeda N and Noguchi S . (2000a). _Int. J. Cancer_, 88, 733–736. * Iwao K, Miyoshi Y, Egawa C,

Ikeda N, Tsukamoto F and Noguchi S . (2000b). _Cancer_, 89, 1732–1738. * Jones PL, Veenstra GJ, Wade PA, Vermaak D, Kass SU, Landsberger N, Strouboulis J and Wolffe AP . (1998). _Nat.

Genet._, 19, 187–191. * Karpf AR and Jones DA . (2002). _Oncogene_, 21, 5496–5503. * Kokura K, Kaul SC, Wadhwa R, Nomura T, Khan MM, Shinagawa T, Yasukawa T, Colmenares C and Ishii S .

(2001). _J. Biol. Chem._, 276, 34115–34121. * Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S and Gustafsson JA . (1996). _Proc. Natl. Acad. Sci. USA_, 93, 5925–5930. * Lewandowski S, Kalita

K and Kaczmarek L . (2002). _FEBS Lett._, 524, 1–5. * Leygue E, Dotzlaw H, Watson PH and Murphy LC . (1998). _Cancer Res._, 58, 3197–3201. * Leygue E, Dotzlaw H, Watson PH and Murphy LC .

(1999). _Cancer Res._, 59, 1175–1179. * Li LC, Yeh CC, Nojima D and Dahiya R . (2000). _Biochem. Biophys. Res. Commun._, 275, 682–689. * Mangelsdorf DJ, Thummel C, Beato M, Herrlich P,

Schutz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P and Evans RM . (1995). _Cell_, 83, 835–839. * Moore JT, McKee DD, Slentz-Kesler K, Moore LB, Jones SA, Horne EL, Su JL, Kliewer

SA, Lehmann JM and Willson TM . (1998). _Biochem. Biophys. Res. Commun._, 247, 75–78. * Nojima D, Li LC, Dharia A, Perinchery G, Ribeiro-Filho L, Yen TS and Dahiya R . (2001). _Cancer_, 92,

2076–2083. * Ogawa S, Inoue S, Watanabe T, Hiroi H, Orimo A, Hosoi T, Ouchi Y and Muramatsu M . (1998a). _Biochem. Biophys. Res. Commun._, 243, 122–126. * Ogawa S, Inoue S, Watanabe T, Orimo

A, Hosoi T, Ouchi Y and Muramatsu M . (1998b). _Nucleic Acids Res._, 26, 3505–3512. * Omoto Y, Kobayashi S, Inoue S, Ogawa S, Toyama T, Yamashita H, Muramatsu M, Gustafsson JA and Iwase H .

(2002). _Eur. J. Cancer_, 38, 380–386. * Palmieri C, Cheng GJ, Saji S, Zelada-Hedman M, Warri A, Weihua Z, Van Noorden S, Wahlstrom T, Coombes RC, Warner M and Gustafsson JA . (2002).

_Endocr. Relat. Cancer_, 9, 1–13. * Sasaki M, Tanaka Y, Perinchery G, Dharia A, Kotcherguina I, Fujimoto S and Dahiya R . (2002). _J. Natl. Cancer Inst._, 94, 384–390. * Skliris GP, Carder

PJ, Lansdown MR and Speirs V . (2001). _Br. J. Cancer_, 84, 1095–1098. * Tate PH and Bird AP . (1993). _Curr. Opin. Genet. Dev._, 3, 226–231. * Tong D, Schuster E, Seifert M, Czerwenka K,

Leodolte S and Zeillinger R . (2002). _Breast Cancer Res. Treat._, 71, 249–255. Download references ACKNOWLEDGEMENTS We thank Laki Buluwela for supplying one of the normal breast epithelial

cell samples. This study was supported by grants from the Swedish Cancer Fund and from KaroBio AB. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Biosciences at Novum,

Karolinska Institutet, Huddinge, S-14157, Sweden Chunyan Zhao, Eva Enmark, Jan-Åke Gustafsson & Karin Dahlman-Wright * Department of Cancer Medicine, Cancer Research-UK labs and Section

of Cancer Cell Biology, Imperial College, London, W12 0NN, UK Eric W-F Lam, Andrew Sunters, Manuela Tamburo De Bella & R Charles Coombes Authors * Chunyan Zhao View author publications

You can also search for this author inPubMed Google Scholar * Eric W-F Lam View author publications You can also search for this author inPubMed Google Scholar * Andrew Sunters View author

publications You can also search for this author inPubMed Google Scholar * Eva Enmark View author publications You can also search for this author inPubMed Google Scholar * Manuela Tamburo

De Bella View author publications You can also search for this author inPubMed Google Scholar * R Charles Coombes View author publications You can also search for this author inPubMed Google

Scholar * Jan-Åke Gustafsson View author publications You can also search for this author inPubMed Google Scholar * Karin Dahlman-Wright View author publications You can also search for

this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Chunyan Zhao. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Zhao, C., Lam,

EF., Sunters, A. _et al._ Expression of estrogen receptor _β_ isoforms in normal breast epithelial cells and breast cancer: regulation by methylation. _Oncogene_ 22, 7600–7606 (2003).

https://doi.org/10.1038/sj.onc.1207100 Download citation * Received: 02 April 2003 * Revised: 08 August 2003 * Accepted: 08 August 2003 * Published: 23 October 2003 * Issue Date: 23 October

2003 * DOI: https://doi.org/10.1038/sj.onc.1207100 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link

is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * ER_β_ * DNA methylation * mRNA isoform * breast

cancer