Skip to main content

Advertisement

SpringerLink
Log in

Effects of exon-deleted estrogen receptor β transcript variants on growth, apoptosis and gene expression of human breast cancer cell lines

  • Preclinical Study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Estrogen receptor β gene codes for a variety of transcript isoforms resulting from alternative splicing, which are expressed both in mammary gland and in breast cancer cells. We studied the function of two exon-deleted ERβ isoforms recently identified by our group in comparison to ERβ1 in regulation of growth, apoptosis and gene expression of two breast cancer cell lines with different ERα status. Overexpression of ERβ1, but not of the exon-deleted variants exerted strong antitumoral effects both on ERα-positive MCF-7 and ERα-negative SK-BR-3 cells. ERβ1 overexpression slowed growth of MCF-7 and SK-BR-3 cells in the absence of E2 and also inhibited E2-triggered growth stimulation of MCF-7 cells, but overexpression of the exon-skipped variants did not affect cell growth. Whereas overexpression of ERβ1 triggered an increased basal and tamoxifen-induced apoptosis of MCF-7 and SK-BR-3 cells, the isoforms ERβδ125 or ERβδ1256 did not affect cellular tamoxifen response. The observed lack of function of the exon-deleted variants in terms of regulation of proliferation was accompanied both by their inability to affect expression of cyclins D1 and A2, p21 (WAF1) and PR and their disability to modulate estrogen response element (ERE) activation. In contrast, our results demonstrating antitumoral effects of ERβ1 on breast cancer cells with different ERα-status support the hypothesis that ERβ is able to exert antitumoral actions both on ERα-positive and -negative breast cancer cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Ali S, Coombes RC (2002) Endocrine-responsive breast cancer and strategies for combating resistance. Nat Rev Cancer 2(2):101–112

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  3. Leygue E, Dotzlaw H, Watson P, Murphy L (1998) Altered estrogen receptor alpha and beta mRNA expression during human breast tumorigenesis. Cancer Res 58:3197–3201

    PubMed  CAS  Google Scholar 

  4. Jarvinen T, 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 156:29–35

    PubMed  CAS  Google Scholar 

  5. Forster C, Makela S, Warri A, Kietz S, Becker D, Hultenby K, Warner M, Gustafsson J (2002) Involvement of estrogen receptor b in terminal differentiation of mammary gland epithelium. PNAS 99:15578–15583

    Article  PubMed  CAS  Google Scholar 

  6. Hall J, McDonnell D (1999) The estrogen receptor betaisoform (ERbeta) of the human estrogen receptor modulates ERalpha transcriptional activity and is a key regulator of the cellular response to estrogens and antiestrogens. Endocrinology 140:5566–5578

    Article  PubMed  CAS  Google Scholar 

  7. Weihua Z, Saji S, Makinen S, Cheng G, Jensen E, Warner M, Gustaffson JA (2000) Estrogen receptor (ER) b a modulator of ERa in the uterus. PNAS 97:5936–5941

    Article  PubMed  CAS  Google Scholar 

  8. Peng B, Lu B, Leygue E, Murphy L (2003) Putative functional characteristics of human estrogen receptor-beta isoforms. J Mol Endocrinol 30:13–29

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  10. Skliris G, Munot K, Bell S, Carder P, Lane S, Horgan K, Lansdown M, Parkes A, Hanby A, Markham A, Speirs V (2003) Reduced expression of oestrogen receptor beta in invasive breast cancer and its re-expression using DNA methyltransferase inhibitors in a cell line model. J Pathol 201:213–220

    Article  PubMed  CAS  Google Scholar 

  11. Dotzlaw H, Leygue E, Watson P, Murphy L (1999) Estrogen receptor-b messenger RNA expression in human breast tumor biopsies: relationship to steroid receptor status and regulation by progestins. Cancer Res 59:529–532

    PubMed  CAS  Google Scholar 

  12. Speirs V (2002) Oestrogen receptor beta in breast cancer: good bad or still too early to tell? J Pathol 197:143–147

    Article  PubMed  CAS  Google Scholar 

  13. Fuqua S, Schiff R, Parra I, Friedrichs W, Su J, McKee D, Slentz-Kesler K, Moore L, Willson T, Moore J (1999) Expression of wild-type estrogen receptor beta and variant isoforms in human breast cancer. Cancer Res 59:5425–5428

    PubMed  CAS  Google Scholar 

  14. Saji S, Omoto Y, Shimizu C, Warner M, Hayashi Y, Horiguchi S, Watanabe T, Hayashi S, Gustafsson J (2002b) Expression of estrogen receptor (ER) (beta)cx protein in ER(alpha)-positive breast cancer: specific correlation with progesterone receptor. Cancer Res 62:4849–4853

    PubMed  CAS  Google Scholar 

  15. Saji S, Omoto Y, Shimizu C, Horiguchi S, Watanabe T, Funata N, Hayash S, Gustafsson J, Toi M (2002a) A clinical impact of assay of estrogen receptor beta cx in breast cancer. Breast Cancer Res Treat 9:303–307

    Google Scholar 

  16. 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):113–118

    Article  PubMed  CAS  Google Scholar 

  17. Murphy L, Cherlet T, Lewis A, Banu Y, Watson P (2003) New insights into estogen receptor function in human breast cancer. Ann Med 35:614–631

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  20. 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):7490–7499

    Article  PubMed  CAS  Google Scholar 

  21. Saji S, Hirose M, Toi M (2005) Clinical significance of estrogen receptor beta in breast cancer. Cancer Chemother Pharmacol 56:21–26

    Article  PubMed  CAS  Google Scholar 

  22. Monroe DG, Secreto FJ, Subramaniam M, Getz BJ, Khosla S, Spelsberg TC (2005) Estrogen receptor alpha and beta heterodimers exert unique effects on estrogen- and tamoxifen-dependent gene expression in human U2OS osteosarcoma cells. Mol Endocrinol 19(6):1555–1568

    Article  PubMed  CAS  Google Scholar 

  23. Osborne C (1998) Steroid hormone receptors in breast cancer management. Breast Cancer Res Treat 51:227–238

    Article  PubMed  CAS  Google Scholar 

  24. Jensen E, Cheng G, Palmieri C, Saji S, Makela S, Van Noorden S, Wahlstrom T, Warner M, Coombes R, Gustafsson J-A (2001) Estrogen receptors and proliferation markers in primary and recurrent breast cancer. PNAS 98:15197–15202

    Article  PubMed  CAS  Google Scholar 

  25. O’Neill P, Davies M, Shaaban A, Innes H, Torevell A, Sibson D, Foster C (2004) Wild-type oestrogen receptor beta (ERbeta1) mRNA and protein expression in Tamoxifen treated post-menopausal breast cancers. Br J Cancer 91:1694–1702

    PubMed  CAS  Google Scholar 

  26. Ogawa S, Inoue S, Watanabe T, Orimo A, Hosoi T, Ouchi Y, Muramatsu M (1998) Molecular cloning and characterization of human estrogen receptor bcx: potential inhibitor of estrogen action in human. Nucleic Acids Res 26:3505–3512

    Article  PubMed  CAS  Google Scholar 

  27. Poola I, Abraham J, Liu A (2002a) Estrogen receptor beta splice variant mRNAs are differentially altered during breast carcinogenesis. J Steroid Biochem Mol Biol 82(2–3):169–179

    Article  PubMed  CAS  Google Scholar 

  28. Price RH Jr, Butler CA, Webb P, Uht R, Kushner P, Handa RJ (2001) A splice variant of estrogen receptor beta missing exon 3 displays altered subnuclear localization and capacity for transcriptional activation. Endocrinology 142(5):2039–2049

    Article  PubMed  CAS  Google Scholar 

  29. Herynk MH, Fuqua SA (2004) Estrogen receptor mutations in human disease. Endocr Rev 25(6):869–898

    Article  PubMed  CAS  Google Scholar 

  30. Price RH Jr, Lorenzon N, Handa RJ (2000) Differential expression of estrogen receptor beta splice variants in rat brain: identification and characterization of a novel variant missing exon 4. Mol Brain Res 80(2):260–268

    Article  PubMed  CAS  Google Scholar 

  31. Speirs V, Adams IP, Walton DS, Atkin SL (2000) Identification of wild-type and exon 5 deletion variants of estrogen receptor beta in normal human mammary gland. J Clin Endocrinol Metab 85(4):1601–1605

    Article  PubMed  CAS  Google Scholar 

  32. Zhao C, Toresson G, Xu L, Koehler KF, Gustafsson JA, Dahlman-Wright K (2005) Mouse estrogen receptor beta isoforms exhibit differences in ligand selectivity and coactivator recruitment. Biochemistry 44(22):7936–7944

    Article  PubMed  CAS  Google Scholar 

  33. Sierens JE, Scobie GA, Wilson J, Saunders PT (2004) Cloning of oestrogen receptor beta from Old and New World primates: identification of splice variants and functional analysis. J Mol Endocrinol 32(3):703–718

    Article  PubMed  CAS  Google Scholar 

  34. Treeck O, Pfeiler G, Horn F, Federhofer B, Houlihan H, Vollmer A, Ortmann O (2007) Novel estrogen receptor beta transcript variants identified in human breast cancer cells affect cell growth and apoptosis of COS-1 cells. Mol Cell Endocrinol 264:50–60

    Article  PubMed  CAS  Google Scholar 

  35. Ståhlberg A, Åman P, Ridell B, Mostad P, Kubista M (2003) A quantitative real-time PCR method for detection of B-Lymphocyte monoclonality by comparison of kappa and lambda Immunoglobulin Light Chain Expression. Clin Chem 49(1):51–59

    Article  PubMed  Google Scholar 

  36. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2^( -delta delta Ct ) method. Methods 25(4):402–408

    Article  PubMed  CAS  Google Scholar 

  37. Cheng J, Lee EJ, Madison LD, Lazennec G (2004) Expression of estrogen receptor beta in prostate carcinoma cells inhibits invasion and proliferation and triggers apoptosis. FEBS Lett 566(1–3):169–172

    Article  PubMed  CAS  Google Scholar 

  38. Helguero LA, Faulds MH, Gustafsson JA, Haldosen LA (2005) Estrogen receptors alpha (ERalpha) and beta (ERbeta) differentially regulate proliferation and apoptosis of the normal murine mammary epithelial cell line HC11. Oncogene 24(44):6605–6616

    Article  PubMed  CAS  Google Scholar 

  39. Treeck O, Pfeiler G, Mitter D, Lattrich C, Piendl G, Ortmann O (2007) Estrogen receptor β1 exerts antitumoral effects on SK-OV-3 ovarian cancer cells. J Endocrinol 193(3):421–433

    Article  PubMed  CAS  Google Scholar 

  40. Vladusic EA, Hornby AE, Guerra-Vladusic FK, Lakins J, Lupu R (2000) Expression and regulation of estrogen receptor beta in human breast tumors and cell lines. Oncol Rep 7(1):157–167

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  42. Guerini V, Sau D, Scaccianoce E, Rusmini P, Ciana P, Maggi A, Martini PG, Katzenellenbogen BS, Martini L, Motta M, Poletti A (2005) The androgen derivative 5alpha-androstane-3beta, 17beta-diol inhibits prostate cancer cell migration through activation of the estrogen receptor beta subtype. Cancer Res 65(12):5445–5453

    Article  PubMed  CAS  Google Scholar 

  43. Lu B, Leygue E, Dotzlaw H, Murphy LJ, Murphy LC, Watson PH (1998) Estrogen receptor-beta mRNA variants in human and murine tissues. Mol Cell Endocrinol 138(1–2):199–203

    Article  PubMed  CAS  Google Scholar 

  44. Vladusic EA, Hornby AE, Guerra-Vladusic FK, Lupu R (1998) Expression of estrogen receptor beta messenger RNA variant in breast cancer. Cancer Res 58(2):210–214

    PubMed  CAS  Google Scholar 

  45. Poola I, Abraham J, Baldwin K (2002b) Identification of ten exon deleted ERbeta mRNAs in human ovary, breast, uterus and bone tissues: alternate splicing pattern of estrogen receptor beta mRNA is distinct from that of estrogen receptor alpha. FEBS Lett 516(1–3):133–138

    Article  PubMed  CAS  Google Scholar 

  46. Omoto Y, Eguchi H, Yamamoto-Yamaguchi Y, Hayashi S (2003) Estrogen receptor (ER) beta1 and ERbetacx/beta2 inhibit ERalpha function differently in breast cancer cell line MCF7. Oncogene 22:5011–5020

    Article  PubMed  CAS  Google Scholar 

  47. Paruthiyil S, Parmar H, Kerekatte V, Cunha GR, Firestone GL, Leitman DC (2004) Estrogen receptor beta inhibits human breast cancer cell proliferation and tumor formation by causing a G2 cell cycle arrest. Cancer Res 64(1):423–428

    Article  PubMed  CAS  Google Scholar 

  48. Hartman J, Lindberg K, Morani A, Inzunza J, Strom A, Gustafsson JA (2006) Estrogen receptor beta inhibits angiogenesis and growth of T47D breast cancer xenografts. Cancer Res 66(23):11207–11213

    Article  PubMed  CAS  Google Scholar 

  49. Strom A, Hartman J, Foster JS, Kietz S, Wimalasena J, Gustafsson JA (2004) Estrogen receptor beta inhibits 17beta-estradiol-stimulated proliferation of the breast cancer cell line T47D. Proc Natl Acad Sci USA 101(6):1566–1571

    Article  PubMed  CAS  Google Scholar 

  50. Lindberg MK, Moverare S, Skrtic S, Gao H, Dahlman-Wright K, Gustafsson JA, Ohlsson C (2003) Estrogen receptor (ER)-beta reduces ERalpha-regulated gene transcription,supporting a “ying yang” relationship between ERalpha and ERbeta in mice. Mol Endocrinol 17(2):203–208

    Article  PubMed  CAS  Google Scholar 

  51. Stossi F, Barnett DH, Frasor J, Komm B, Lyttle CR, Katzenellenbogen BS (2004) Transcriptional profiling of estrogen-regulated gene expression via estrogen receptor (ER) alpha or ERbeta in human osteosarcoma cells: distinct and common target genes for these receptors. Endocrinology 145(7):3473–3486

    Article  PubMed  CAS  Google Scholar 

  52. Chang EC, Frasor J, Komm B, Katzenellenbogen BS (2006) Impact of estrogen receptor beta on gene networks regulated by estrogen receptor alpha in breast cancer cells. Endocrinology 147(10):4831–4842

    Article  PubMed  CAS  Google Scholar 

  53. Lin CY, Ström A, Li Kong S, Kietz S, Thomsen JS, Tee JB, Vega VB, Miller LD, Smeds J, Bergh J, Gustafsson JA, Liu ET, (2007) Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer. Breast Cancer Res 9(2):R25

    Article  PubMed  CAS  Google Scholar 

  54. Carroll JS, Swarbrick A, Musgrove EA, Sutherland RL (2002) Mechanisms of growth arrest by c-myc antisense oligonucleotides in MCF-7 breast cancer cells: implications for the antiproliferative effects of antiestrogens. Cancer Res 62:3126–3131

    PubMed  CAS  Google Scholar 

  55. Watts CK, Sweeney KJ, Warlters A, Musgrove EA, Sutherland RL (1994) Antiestrogen regulation of cell cycle progression and cyclin D1 gene expression in MCF-7 human breast cancer cells. Breast Cancer Res Treat 31:95–105

    Article  PubMed  CAS  Google Scholar 

  56. Foster JS, Henley DC, Bukovsky A, Seth P, Wimalasena J (2001) Multifaceted regulation of cell cycle progression by estrogen: regulation of Cdk inhibitors and Cdc25A independent of cyclin D1-Cdk4 function. Mol Cell Biol 21:794–810

    Article  PubMed  CAS  Google Scholar 

  57. Liu MM, Albanese C, Anderson CM, Hilty K, Webb P, Uht RM, Price RH Jr, Pestell RG, Kushner PJ (2002) Opposing action of estrogen receptors alpha and beta on cyclin D1 gene expression. J Biol Chem 277(27):24353–24360

    Article  PubMed  CAS  Google Scholar 

  58. Hodges LC, Cook JD, Lobenhofer EK, Li L, Bennett L, Bushel PR, Aldaz CM, Afshari CA, Walker CL (2003) Tamoxifen functions as a molecular agonist inducing cell cycle-associated genes in breast cancer cells. Mol Cancer Res 1:300–311

    PubMed  CAS  Google Scholar 

  59. Hui R, Finney GL, Carroll JS, Lee CS, Musgrove EA, Sutherland RL (2002) Constitutive overexpression of cyclin D1 but not cyclin E confers acute resistance to antiestrogens in T-47D breast cancer cells. Cancer Res 62:6916–6923

    PubMed  CAS  Google Scholar 

  60. Doisneau-Sixou SF, Sergio CM, Carroll JS, Hui R, Musgrove EA, Sutherland RL (2003) Estrogen and antiestrogen regulation of cell cycle progression in breast cancer cells. Endocr Relat Cancer 10:179–186

    Article  PubMed  CAS  Google Scholar 

  61. Thomas TJ, Faaland CA, Adhikarakunnathu S, Watkins LF, Thomas T (1998) Induction of p21 (CIP1/WAF1/SID1) by estradiol in a breast epithelial cell line transfected with the recombinant estrogen receptor gene: a possible mechanism for a negative regulatory role of estradiol. Breast Cancer Res Treat 47(2):181–193

    Article  PubMed  CAS  Google Scholar 

  62. 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):34–38

    Article  CAS  Google Scholar 

  63. Gruvberger-Saal SK, Bendahl PO, Saal LH, Laakso M, Hegardt C, Eden P, Peterson C, Malmström P, Isola J, Borg A, Fernö M (2007) Estrogen receptor beta expression is associated with tamoxifen response in ERalpha-negative breast carcinoma. Clin Cancer Res 13(7):1987–1994

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank Angelika Vollmer, Gerhard Piendl and Helena Houlihan for expert technical assistance.

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, University of Regensburg, Landshuter Str. 65, 93053, Regensburg, Germany

    Oliver Treeck, Ingolf Juhasz-Boess, Claus Lattrich, Felicitas Horn, Regina Goerse & Olaf Ortmann

Authors
  1. Oliver Treeck
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ingolf Juhasz-Boess
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Claus Lattrich
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Felicitas Horn
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Regina Goerse
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Olaf Ortmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Oliver Treeck.

Electronic supplementary material

Below is the link to the electronic supplementary material

ESM 1 (PDF 87 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Treeck, O., Juhasz-Boess, I., Lattrich, C. et al. Effects of exon-deleted estrogen receptor β transcript variants on growth, apoptosis and gene expression of human breast cancer cell lines. Breast Cancer Res Treat 110, 507–520 (2008). https://doi.org/10.1007/s10549-007-9749-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-007-9749-7

Keywords

Search

Navigation