Advertisement

Breast Cancer Research and Treatment

, Volume 126, Issue 1, pp 27–36 | Cite as

The ErbB3 binding protein EBP1 regulates ErbB2 protein levels and tamoxifen sensitivity in breast cancer cells

  • Yan Lu
  • Hua Zhou
  • Wantao Chen
  • Yuexing Zhang
  • Anne W. Hamburger
Preclinical study

Abstract

The ErbB2/3 heterodimer plays a critical role in breast cancer progression and in the development of endocrine resistance. EBP1, an ErbB3 binding protein, inhibits HRG-stimulated breast cancer growth, decreases ErbB2 protein levels and contributes to tamoxifen sensitivity. We report here that ectopic expression of EBP1 in Estrogen Receptor (ER) positive breast cancers that express ErbB2 at both high and low levels decreased ErbB2 protein levels. ErbB2 protein expression was also increased in mammary glands of Ebp1 knock out mice. To define the mechanism of ErbB2 down regulation, we examined the effects of EBP1 on ErbB2 mRNA levels, transcription of the ErbB2 gene and ErbB2 protein stability. We found that ectopic expression of EBP1 decreased steady state levels of endogenous ErbB2 mRNA in all cell lines tested. EBP1 overexpression decreased the activity of an ErbB2 promoter reporter in cells which overxpress ErbB2. However, reporter activity was unchanged or increased in cells which express low endogenous levels of ErbB2. We also found that ectopic expression of EBP1 accelerated ErbB2 protein degradation and enhanced ErbB2 ubiquitination in cells which express both low and high levels of ErbB2. Treatment with proteasome inhibitors prevented this decrease in ErbB2 protein levels. Ablation of EBP1 expression led to tamoxifen resistance that was abrogated by inhibition of ErbB2 activity. These results suggest that EBP1 inhibits expression of ErbB2 protein levels by multiple mechanisms and that EBP1’s effects on tamoxifen sensitivity are mediated in part by its ability to modulate ErbB2 levels.

Keywords

EBP1 ErbB2 Tamoxifen Breast cancer 

Notes

Acknowledgments

This study was supported by NIH grants R01 CA76047, RC1 CA145066-01 and DOD W81XWH-08-1-0560 (to AWH), and by the Project of the Shanghai Science and Technology Committee (08JC1414400, 08JC1404800) and the Shanghai Leading Academic Discipline Project (S30206). We thank Dr. Kay Huebner, Ohio State University Cancer Center, and Dr. Helen Hurst for the ErbB2 promoter plasmid and Dr. Yossi Yarden for the ErbB2 expression construct.

References

  1. 1.
    Holbro T, Civenni G, Hynes NE (2003) The ErbB receptors and their role in cancer progression. Exp Cell Res 284:99–110CrossRefPubMedGoogle Scholar
  2. 2.
    Hynes NE, Lane HA (2005) ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer 5:341–354CrossRefPubMedGoogle Scholar
  3. 3.
    Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, Levin WJ, Stuart SG, Udove J, Ullrich A (1989) Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244:707–712CrossRefPubMedGoogle Scholar
  4. 4.
    Lemoine NR, Barnes DM, Hollywood DP, Hughes CM, Smith P, Dublin E, Prigent SA, Gullick WJ, Hurst HC (1992) Expression of the ERBB3 gene product in breast cancer. Br J Cancer 66:1116–1121CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Wiseman SM, Makretsov N, Nielsen TO, Gilks B, Yorida E, Cheang M, Turbin D, Gelmon K, Huntsman DG (2005) Coexpression of the type 1 growth factor receptor family members HER-1, HER-2, and HER-3 has a synergistic negative prognostic effect on breast carcinoma survival. Cancer 103:1770–1777CrossRefPubMedGoogle Scholar
  6. 6.
    Sergina NV, Rausch M, Wang D, Blair J, Hann B, Shokat KM, Moasser MM (2007) Escape from HER-family tyrosine kinase inhibitor therapy by the kinase-inactive HER3. Nature 445:437–441CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Pinkas-Kramarski R, Soussan L, Waterman H, Levkowitz G, Alroy I, Klapper L, Lavi S, Seger R, Ratzkin BJ, Sela M, Yarden Y (1996) Diversification of Neu differentiation factor and epidermal growth factor signaling by combinatorial receptor interactions. EMBO J 15:2452–2467PubMedPubMedCentralGoogle Scholar
  8. 8.
    Holbro T, Beerli RR, Maurer F, Koziczak M, Barbas CF III, Hynes NE (2003) The ErbB2/ErbB3 heterodimer functions as an oncogenic unit: ErbB2 requires ErbB3 to drive breast tumor cell proliferation. Proc Natl Acad Sci USA 100:8933–8938CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    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
  10. 10.
    Piccart-Gebhart MJ, Procter M, Leyland-Jones B, Goldhirsch A, Untch M, Smith I, Gianni L, Baselga J, Bell R, Jackisch C, Cameron D, Dowsett M, Barrios CH, Steger G, Huang CS, Andersson M, Inbar M, Lichinitser M, Lang I, Nitz U, Iwata H, Thomssen C, Lohrisch C, Suter TM, Ruschoff J, Suto T, Greatorex V, Ward C, Straehle C, McFadden E, Dolci MS, Gelber RD (2005) Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. N Engl J Med 353:1659–1672CrossRefPubMedGoogle Scholar
  11. 11.
    Pietras RJ, Arboleda J, Reese DM, Wongvipat N, Pegram MD, Ramos L, Gorman CM, Parker MG, Sliwkowski MX, Slamon DJ (1995) HER-2 tyrosine kinase pathway targets estrogen receptor and promotes hormone-independent growth in human breast cancer cells. Oncogene 10:2435–2446PubMedGoogle Scholar
  12. 12.
    Benz CC, Scott GK, Sarup JC, Johnson RM, Tripathy D, Coronado E, Shepard HM, Osborne CK (1992) Estrogen-dependent, tamoxifen-resistant tumorigenic growth of MCF-7 cells transfected with HER2/neu. Breast Cancer Res Treat 24:85–95CrossRefPubMedGoogle Scholar
  13. 13.
    Kraus MH, Issing W, Miki T, Popescu NC, Aaronson SA (1989) Isolation and characterization of ERBB3, a third member of the ERBB/epidermal growth factor receptor family: evidence for overexpression in a subset of human mammary tumors. Proc Natl Acad Sci USA 86:9193–9197CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Plowman GD, Whitney GS, Neubauer MG, Green JM, McDonald VL, Todaro GJ, Shoyab M (1990) Molecular cloning and expression of an additional epidermal growth factor receptor-related gene. Proc Natl Acad Sci USA 87:4905–4909CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Yoo JY, Wang XW, Rishi AK, Lessor T, Xia XM, Gustafson TA, Hamburger AW (2000) Interaction of the PA2G4 (EBP1) protein with ErbB-3 and regulation of this binding by heregulin. Br J Cancer 82:683–690CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Lessor TJ, Yoo JY, Xia X, Woodford N, Hamburger AW (2000) Ectopic expression of the ErbB-3 binding protein ebp1 inhibits growth and induces differentiation of human breast cancer cell lines. J Cell Physiol 183:321–329CrossRefPubMedGoogle Scholar
  17. 17.
    Xia X, Cheng A, Lessor T, Zhang Y, Hamburger AW (2001) Ebp1, an ErbB-3 binding protein, interacts with Rb and affects Rb transcriptional regulation. J Cell Physiol 187:209–217CrossRefPubMedGoogle Scholar
  18. 18.
    Zhang YX, Woodford N, Xia XM, Hamburger AW (2003) Repression of E2F1-mediated transcription by the ErbB3 binding protein Ebp1 involves histone deacetylases. Nucl Acids Res 31:2168–2177CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Zhang Y, Hamburger AW (2004) Heregulin regulates the ability of the ErbB3-binding protein Ebp1 to bind E2F promoter elements and repress E2F-mediated transcription. J Biol Chem 279:26126–26133CrossRefPubMedGoogle Scholar
  20. 20.
    Zhang Y, Akinmade D, Hamburger AW (2005) The ErbB3 binding protein Ebp1 interacts with Sin3A to repress E2F1 and AR-mediated transcription. Nucl Acids Res 33:6024–6033CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Zhang Y, Akinmade D, Hamburger AW (2008) Inhibition of heregulin mediated MCF-7 breast cancer cell growth by the ErbB3 binding protein EBP1. Cancer Lett 265:298–306CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Zhang Y, Linn D, Liu Z, Melamed J, Tavora F, Young CY, Burger AM, Hamburger AW (2008) EBP1, an ErbB3-binding protein, is decreased in prostate cancer and implicated in hormone resistance. Mol Cancer Ther 7:3176–3186CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Zhang JS, Gong A, Cheville JC, Smith DI, Young CY (2005) AGR2, an androgen-inducible secretory protein overexpressed in prostate cancer. Genes Chromosomes Cancer 43:249–259CrossRefPubMedGoogle Scholar
  24. 24.
    Zhang Y, Wang XW, Jelovac D, Nakanishi T, Yu MH, Akinmade D, Goloubeva O, Ross DD, Brodie A, Hamburger AW (2005) The ErbB3-binding protein Ebp1 suppresses androgen receptor-mediated gene transcription and tumorigenesis of prostate cancer cells. Proc Natl Acad Sci USA 102:9890–9895CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Johnson DG, Ohtani K, Nevins JR (1994) Autoregulatory control of E2F1 expression in response to positive and negative regulators of cell cycle progression. Genes Dev 8:1514–1525CrossRefPubMedGoogle Scholar
  26. 26.
    Zhang Y, Lu Y, Zhou H, Lee M, Liu Z, Hassel BA, Hamburger AW (2008) Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice. BMC Cell Biol 9:69CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Xu W, Marcu M, Yuan X, Mimnaugh E, Patterson C, Neckers L (2002) Chaperone-dependent E3 ubiquitin ligase CHIP mediates a degradative pathway for c-ErbB2/Neu. Proc Natl Acad Sci USA 99:12847–12852CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Liu Z, Oh SM, Okada M, Liu X, Cheng D, Peng J, Brat DJ, Sun SY, Zhou W, Gu W, Ye K (2009) Human BRE1 is an E3 ubiquitin ligase for Ebp1 tumor suppressor. Mol Biol Cell 20:757–768CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Klapper LN, Waterman H, Sela M, Yarden Y (2000) Tumor-inhibitory antibodies to HER-2/ErbB-2 may act by recruiting c-Cbl and enhancing ubiquitination of HER-2. Cancer Res 60:3384–3388PubMedGoogle Scholar
  30. 30.
    Monie TP, Perrin AJ, Birtley JR, Sweeney TR, Karakasiliotis I, Chaudhry Y, Roberts LO, Matthews S, Goodfellow IG, Curry S (2007) Structural insights into the transcriptional and translational roles of Ebp1. EMBO J 26:3936–3944CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Yamada H, Mori H, Momoi H, Nakagawa Y, Ueguchi C, Mizuno T (1994) A fission yeast gene encoding a protein that preferentially associates with curved DNA. Yeast 10:883–894CrossRefPubMedGoogle Scholar
  32. 32.
    Radomski N, Jost E (1995) Molecular cloning of a murine cDNA encoding a novel protein, p38-2G4, which varies with the cell cycle. Exp Cell Res 220:434–445CrossRefPubMedGoogle Scholar
  33. 33.
    Squatrito M, Mancino M, Sala L, Draetta GF (2006) Ebp1 is a dsRNA-binding protein associated with ribosomes that modulates eIF2alpha phosphorylation. Biochem Biophys Res Commun 344:859–868CrossRefPubMedGoogle Scholar
  34. 34.
    Squatrito M, Mancino M, Donzelli M, Areces LB, Draetta GF (2004) EBP1 is a nucleolar growth-regulating protein that is part of pre-ribosomal ribonucleoprotein complexes. Oncogene 23:4454–4465CrossRefPubMedGoogle Scholar
  35. 35.
    Bose SK, Sengupta TK, Bandyopadhyay S, Spicer EK (2006) Identification of Ebp1 as a component of cytoplasmic bcl-2 mRNP (messenger ribonucleoprotein particle) complexes. Biochem J 396:99–107CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Pilipenko EV, Pestova TV, Kolupaeva VG, Khitrina EV, Poperechnaya AN, Agol VI, Hellen CU (2000) A cell cycle-dependent protein serves as a template-specific translation initiation factor. Genes Dev 14:2028–2045PubMedPubMedCentralGoogle Scholar
  37. 37.
    Ahn JY, Liu X, Liu Z, Pereira L, Cheng D, Peng J, Wade PA, Hamburger AW, Ye K (2006) Nuclear Akt associates with PKC-phosphorylated Ebp1, preventing DNA fragmentation by inhibition of caspase-activated DNase. EMBO J 25:2083–2095CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Okada M, Jang SW, Ye K (2007) Ebp1 association with nucleophosmin/B23 is essential for regulating cell proliferation and suppressing apoptosis. J Biol Chem 282:36744–36754CrossRefPubMedGoogle Scholar
  39. 39.
    Akinmade D, Lee M, Zhang Y, Hamburger AW (2007) Ebp1-mediated inhibition of cell growth requires serine 363 phosphorylation. Int J Oncol 31:851–858PubMedGoogle Scholar
  40. 40.
    Liu Z, Liu X, Nakayama KI, Nakayama K, Ye K (2007) Protein kinase C-delta phosphorylates Ebp1 and prevents its proteolytic degradation, enhancing cell survival. J Neurochem 100:1278–1288CrossRefPubMedGoogle Scholar
  41. 41.
    Qin HR, Iliopoulos D, Nakamura T, Costinean S, Volinia S, Druck T, Sun J, Okumura H, Huebner K (2007) Wwox suppresses prostate cancer cell growth through modulation of ErbB2-mediated androgen receptor signaling. Mol Cancer Res 5:957–965CrossRefPubMedGoogle Scholar
  42. 42.
    Laederich MB, Funes-Duran M, Yen L, Ingalla E, Wu X, Carraway KL III, Sweeney C (2004) The leucine-rich repeat protein LRIG1 is a negative regulator of ErbB family receptor tyrosine kinases. J Biol Chem 279:47050–47056CrossRefPubMedGoogle Scholar
  43. 43.
    Liu Z, Ahn JY, Liu X, Ye K (2006) Ebp1 isoforms distinctively regulate cell survival and differentiation. Proc Natl Acad Sci USA 103:10917–10922CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Hurtado A, Holmes KA, Geistlinger TR, Hutcheson IR, Nicholson RI, Brown M, Jiang J, Howat WJ, Ali S, Carroll JS (2008) Regulation of ERBB2 by oestrogen receptor-PAX2 determines response to tamoxifen. Nature 456:663–666CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Yan Lu
    • 1
    • 2
  • Hua Zhou
    • 1
    • 3
  • Wantao Chen
    • 2
  • Yuexing Zhang
    • 1
    • 3
  • Anne W. Hamburger
    • 1
    • 3
  1. 1.Greenebaum Cancer CenterUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Department of Oral and Maxillofacial SurgeryShanghai Jiaotong University School of MedicineShanghaiChina
  3. 3.Department of PathologyUniversity of Maryland School of MedicineBaltimoreUSA

Personalised recommendations