Breast Cancer Research and Treatment

, Volume 123, Issue 1, pp 87–96

Role of GPR30 in the mechanisms of tamoxifen resistance in breast cancer MCF-7 cells

Preclinical study

Abstract

Tamoxifen is the most frequently used anti-hormonal drug for treatment of women with hormone-dependent breast cancer. The aim of this study is to investigate the mechanism of tamoxifen resistance and the impact of the new estrogen G-protein coupled receptor (GPR30). MCF-7 cells were continuously exposed to tamoxifen for 6 months to induce resistance to the inhibitory effect of tamoxifen. These tamoxifen-resistant cells (TAM-R) exhibited enhanced sensitivity to 17-ß-estradiol and GPR30 agonist, G1, when compared to the parental cells. In TAM-R cells, tamoxifen was able to stimulate the cell growth and MAPK phosphorylation. These effects were abolished by EGFR inhibitor AG1478, GPR30 anti-sense oligonucleotide, and the selective c-Src inhibitor PP2. Only EGFR basal expression was slightly elevated in the TAM-R cells, whereas GPR30 expression and the basal phosphorylation of Akt and MAPK remained unchanged when compared to the parental cells. Interestingly, estrogen treatment significantly increased GPR30 translocation to the cell surface, which was stronger in TAM-R cells. Continuous treatment of MCF-7 cells with GPR30 agonist G1 mimics the long-term treatment with tamoxifen and increases drastically its agonistic activity. This data suggests the important role of GPR30/EGFR receptor signaling in the development of tamoxifen resistance. The inhibition of this pathway is a valid option to improve anti-hormone response in breast cancer.

Keywords

GPR30 Breast cancer Tamoxifen resistance Estrogen 

References

  1. 1.
    Tamoxifen for early breast cancer: an overview of the randomised trials (1998) Early breast cancer trialists’ collaborative group. Lancet 351:1451–1467CrossRefGoogle Scholar
  2. 2.
    Powles TJ, Ashley S, Tidy A, Smith IE, Dowsett M (2007) Twenty-year follow-up of the Royal Marsden randomized, double-blinded tamoxifen breast cancer prevention trial. J Natl Cancer Inst 99:283–290CrossRefPubMedGoogle Scholar
  3. 3.
    Kurebayashi J (2005) Resistance to endocrine therapy in breast cancer. Cancer Chemother Pharmacol 56(Suppl 1):39–46CrossRefPubMedGoogle Scholar
  4. 4.
    Nicholson RI, McClelland RA, Finlay P, Eaton CL, Gullick WJ, Dixon AR, Robertson JF, Ellis IO, Blamey RW (1993) Relationship between EGF-R, c-erbB-2 protein expression and Ki67 immunostaining in breast cancer and hormone sensitivity. Eur J Cancer 29A:1018–1023CrossRefPubMedGoogle Scholar
  5. 5.
    Wright C, Nicholson S, Angus B, Sainsbury JR, Farndon J, Cairns J, Harris AL, Horne CH (1992) Relationship between c-erbB-2 protein product expression and response to endocrine therapy in advanced breast cancer. Br J Cancer 65:118–121PubMedGoogle Scholar
  6. 6.
    Hutcheson IR, Knowlden JM, Madden TA, Barrow D, Gee JM, Wakeling AE, Nicholson RI (2003) Oestrogen receptor-mediated modulation of the EGFR/MAPK pathway in tamoxifen-resistant MCF-7 cells. Breast Cancer Res Treat 81:81–93CrossRefPubMedGoogle Scholar
  7. 7.
    Rae JM, Johnson MD (2005) What does an orphan G-protein-coupled receptor have to do with estrogen? Breast Cancer Res 7:243–244CrossRefPubMedGoogle Scholar
  8. 8.
    Prossnitz ER, Oprea TI, Sklar LA, Arterburn JB (2008) The ins and outs of GPR30: a transmembrane estrogen receptor. J Steroid Biochem Mol Biol 109:350–353CrossRefPubMedGoogle Scholar
  9. 9.
    Filardo EJ (2002) Epidermal growth factor receptor (EGFR) transactivation by estrogen via the G-protein-coupled receptor, GPR30: a novel signaling pathway with potential significance for breast cancer. J Steroid Biochem Mol Biol 80:231–238CrossRefPubMedGoogle Scholar
  10. 10.
    Ignatov A, Lintzel J, Kreienkamp HJ, Schaller HC (2003) Sphingosine-1-phosphate is a high-affinity ligand for the G protein-coupled receptor GPR6 from mouse and induces intracellular Ca2+ release by activating the sphingosine-kinase pathway. Biochem Biophys Res Commun 311:329–336CrossRefPubMedGoogle Scholar
  11. 11.
    Ignatov A, Robert J, Gregory-Evans C, Schaller HC (2006) RANTES stimulates Ca2+ mobilization and inositol trisphosphate (IP3) formation in cells transfected with G protein-coupled receptor 75. Br J Pharmacol 149:490–497CrossRefPubMedGoogle Scholar
  12. 12.
    Jordan NJ, Gee JM, Barrow D, Wakeling AE, Nicholson RI (2004) Increased constitutive activity of PKB/Akt in tamoxifen resistant breast cancer MCF-7 cells. Breast Cancer Res Treat 87:167–180CrossRefPubMedGoogle Scholar
  13. 13.
    Osborne CK, Shou J, Massarweh S, Schiff R (2005) Crosstalk between estrogen receptor and growth factor receptor pathways as a cause for endocrine therapy resistance in breast cancer. Clin Cancer Res 11:865s–870sPubMedGoogle Scholar
  14. 14.
    Revankar CM, Cimino DF, Sklar LA, Arterburn JB, Prossnitz ER (2005) A transmembrane intracellular estrogen receptor mediates rapid cell signaling. Science 307:1625–1630CrossRefPubMedGoogle Scholar
  15. 15.
    Thomas P, Pang Y, Filardo EJ, Dong J (2005) Identity of an estrogen membrane receptor coupled to a G protein in human breast cancer cells. Endocrinology 146:624–632CrossRefPubMedGoogle Scholar
  16. 16.
    Filardo EJ, Quinn JA, Bland KI, Frackelton AR Jr (2000) Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF. Mol Endocrinol 14:1649–1660CrossRefPubMedGoogle Scholar
  17. 17.
    Maggiolini M, Vivacqua A, Fasanella G, Recchia AG, Sisci D, Pezzi V, Montanaro D, Musti AM, Picard D, Ando S (2004) The G protein-coupled receptor GPR30 mediates c-fos up-regulation by 17 beta-estradiol and phytoestrogens in breast cancer cells. J Biol Chem 279:27008–27016CrossRefPubMedGoogle Scholar
  18. 18.
    Vivacqua A, Bonofiglio D, Recchia AG, Musti AM, Picard D, Ando S, Maggiolini M (2006) The G protein-coupled receptor GPR30 mediates the proliferative effects induced by 17 beta-estradiol and hydroxytamoxifen in endometrial cancer cells. Mol Endocrinol 20:631–646CrossRefPubMedGoogle Scholar
  19. 19.
    Vivacqua A, Bonofiglio D, Albanito L, Madeo A, Rago V, Carpino A, Musti AM, Picard D, Ando S, Maggiolini M (2006) 17 beta-estradiol, genistein, and 4-hydroxytamoxifen induce the proliferation of thyroid cancer cells through the g protein-coupled receptor GPR30. Mol Pharmacol 70:1414–1423CrossRefPubMedGoogle Scholar
  20. 20.
    Levin ER (2009) G protein-coupled receptor 30: estrogen receptor or collaborator? Endocrinology 150:1563–1565CrossRefPubMedGoogle Scholar
  21. 21.
    Leblanc K, Sexton E, Parent S, Belanger G, Dery MC, Boucher V, Asselin E (2007) Effects of 4-hydroxytamoxifen, raloxifene and ICI 182 780 on survival of uterine cancer cell lines in the presence and absence of exogenous estrogens. Int J Oncol 30:477–487PubMedGoogle Scholar
  22. 22.
    Fan P, Yue W, Wang JP, Aiyar S, Li Y, Kim TH, Santen RJ (2009) Mechanisms of resistance to structurally diverse antiestrogens differ under premenopausal and postmenopausal conditions: evidence from in vitro breast cancer cell models. Endocrinology 150:2036–2045CrossRefPubMedGoogle Scholar
  23. 23.
    Bedard PL, Freedman OC, Howell A, Clemons M (2008) Overcoming endocrine resistance in breast cancer: are signal transduction inhibitors the answer? Breast Cancer Res Treat 108:307–317CrossRefPubMedGoogle Scholar
  24. 24.
    Levin ER (2003) Bidirectional signaling between the estrogen receptor and the epidermal growth factor receptor. Mol Endocrinol 17:309–317CrossRefPubMedGoogle Scholar
  25. 25.
    Filardo EJ, Graeber CT, Quinn JA, Resnick MB, Giri D, DeLellis RA, Steinhoff MM, Sabo E (2006) Distribution of GPR30, a seven membrane-spanning estrogen receptor, in primary breast cancer and its association with clinicopathologic determinants of tumor progression. Clin Cancer Res 12:6359–6366CrossRefPubMedGoogle Scholar
  26. 26.
    Vivacqua A, Lappano R, De Marco P, Sisci D, Aquila S, De Amicis F, Fuqua SA, Ando S, Maggiolini M (2009) G Protein-coupled receptor 30 expression is up-regulated by EGF and TGFα in estrogen receptor α-positive cancer cells. Mol Endocrinol. doi:10.1210/me.2009-0120
  27. 27.
    Fan P, Wang J, Santen RJ, Yue W (2007) Long-term treatment with tamoxifen facilitates translocation of estrogen receptor alpha out of the nucleus and enhances its interaction with EGFR in MCF-7 breast cancer cells. Cancer Res 67:1352–1360CrossRefPubMedGoogle Scholar
  28. 28.
    Gee JM, Harper ME, Hutcheson IR, Madden TA, Barrow D, Knowlden JM, McClelland RA, Jordan N, Wakeling AE, Nicholson RI (2003) The antiepidermal growth factor receptor agent gefitinib (ZD1839/Iressa) improves antihormone response and prevents development of resistance in breast cancer in vitro. Endocrinology 144:5105–5117CrossRefPubMedGoogle Scholar
  29. 29.
    Shou J, Massarweh S, Osborne CK, Wakeling AE, Ali S, Weiss H, Schiff R (2004) Mechanisms of tamoxifen resistance: increased estrogen receptor-HER2/neu cross-talk in ER/HER2-positive breast cancer. J Natl Cancer Inst 96:926–935CrossRefPubMedGoogle Scholar
  30. 30.
    Gutierrez MC, Detre S, Johnston S, Mohsin SK, Shou J, Allred DC, Schiff R, Osborne CK, Dowsett M (2005) Molecular changes in tamoxifen-resistant breast cancer: relationship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. J Clin Oncol 23:2469–2476CrossRefPubMedGoogle Scholar
  31. 31.
    Razandi M, Pedram A, Park ST, Levin ER (2003) Proximal events in signaling by plasma membrane estrogen receptors. J Biol Chem 278:2701–2712CrossRefPubMedGoogle Scholar
  32. 32.
    deFazio A, Chiew YE, Sini RL, Janes PW, Sutherland RL (2000) Expression of c-erbB receptors, heregulin and oestrogen receptor in human breast cell lines. Int J Cancer 87:487–498CrossRefPubMedGoogle Scholar
  33. 33.
    Wilson MA, Chrysogelos SA (2002) Identification and characterization of a negative regulatory element within the epidermal growth factor receptor gene first intron in hormone-dependent breast cancer cells. J Cell Biochem 85:601–614CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2009

Authors and Affiliations

  1. 1.Department of Obstetrics and GynecologyOtto-von-Guericke UniversityMagdeburgGermany
  2. 2.Department of PathologyOtto-von-Guericke UniversityMagdeburgGermany

Personalised recommendations