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Molecular and Cellular Biochemistry

, Volume 405, Issue 1–2, pp 177–186 | Cite as

The ErbB3-binding protein EBP1 modulates lapatinib sensitivity in prostate cancer cells

  • Smita Awasthi
  • Heather Ezelle
  • Bret A. Hassel
  • Anne W. Hamburger
Article

Abstract

Although ErbB receptors have been implicated in prostate cancer progression, ErbB-directed drugs have not proven effective for prostate cancer treatment. The ErbB3-binding protein EBP1 affects both ErbB2 and androgen receptor signaling, two components of the response to ErbB-targeted therapies. We therefore examined the effects of EBP1 expression on the response to the ErbB1/2 tyrosine kinase inhibitor lapatinib. We found a negative correlation between endogenous EBP1 levels and lapatinib sensitivity in prostate cancer cell lines. We then overexpressed or inhibited expression of EBP1. Silencing EBP1 expression increased lapatinib sensitivity and overexpression of EBP1 increased resistance in androgen-containing media. Androgen depletion resulted in an increased sensitivity of androgen-dependent EBP1 expressing cells to lapatinib, but did not affect the lapatinib sensitivity of hormone resistant cells. However, EBP1 silenced cells were still more sensitive to lapatinib than EBP1-expressing cells in the absence of androgens. The increase in sensitivity to lapatinib following EBP1 silencing was associated with increased ErbB2 levels. In addition, lapatinib treatment increased ErbB2 levels in sensitive cells that express low levels of EBP1, but decreased ErbB2 levels in resistant EBP1-expressing cells. In contrast, ErbB3 and phospho ErbB3 levels were not affected by either changes in EBP1 levels or lapatinib treatment. The production of the ErbB3/4 ligand heregulin was increased in EBP1-silenced cells. EBP1-induced changes in AR levels were not associated with changes in lapatinib sensitivity. These studies suggest that the ability of EBP1 to activate ErbB2 signaling pathways results in increased lapatinib sensitivity.

Keywords

EBP1 Prostate cancer ErbB2 Lapatinib 

Notes

Acknowledgments

This work was supported by NIH Grant 1R01CA138583 (AWH). We thank Dr. M. Lin (University of Nebraska Medical Center) for permission to use the C81 cells and Dr. Yun Qiu (University of Maryland School of Medicine) for providing these cells.

Supplementary material

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Supplementary material 1 (PDF 169 kb)
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Supplementary material 2 (PDF 95 kb)
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Supplementary material 3 (PDF 6 kb)
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Supplementary material 4 (PDF 90 kb)
11010_2015_2409_MOESM5_ESM.docx (27 kb)
Supplementary material 5 (DOCX 26 kb)

References

  1. 1.
    Craft N, Shostak Y, Carey M, Sawyers CL (1999) A mechanism for hormone-independent prostate cancer through modulation of androgen receptor signaling by the HER-2/neu tyrosine kinase. Nat Med 5:280–285CrossRefPubMedGoogle Scholar
  2. 2.
    Yeh S, Lin HK, Kang HY, Thin TH, Lin MF, Chang C (1999) From HER2/Neu signal cascade to androgen receptor and its coactivators: a novel pathway by induction of androgen target genes through MAP kinase in prostate cancer cells. Proc Natl Acad Sci USA 96:5458–5463CrossRefPubMedCentralPubMedGoogle Scholar
  3. 3.
    Koumakpayi IH, Diallo JS, Le PC, Lessard L, Gleave M, Begin LR, Mes-Masson AM, Saad F (2006) Expression and nuclear localization of ErbB3 in prostate cancer. Clin Cancer Res 12:2730–2737CrossRefPubMedGoogle Scholar
  4. 4.
    Mellinghoff IK, Tran C, Sawyers CL (2002) Growth inhibitory effects of the dual ErbB1/ErbB2 tyrosine kinase inhibitor PKI-166 on human prostate cancer xenografts. Cancer Res 62:5254–5259PubMedGoogle Scholar
  5. 5.
    Agus DB, Akita RW, Fox WD, Lewis GD, Higgins B, Pisacane PI, Lofgren JA, Tindell C, Evans DP, Maiese K, Scher HI, Sliwkowski MX (2002) Targeting ligand-activated ErbB2 signaling inhibits breast and prostate tumor growth. Cancer Cell 2:127–137CrossRefPubMedGoogle Scholar
  6. 6.
    Mendoza N, Phillips GL, Silva J, Schwall R, Wickramasinghe D (2002) Inhibition of ligand-mediated HER2 activation in androgen-independent prostate cancer. Cancer Res 62:5485–5488PubMedGoogle Scholar
  7. 7.
    Liu G, Chen YH, Kolesar J, Huang W, Dipaola R, Pins M, Carducci M, Stein M, Bubley GJ, Wilding G (2013) Eastern Cooperative Oncology Group phase II trial of lapatinib in men with biochemically relapsed, androgen dependent prostate cancer. Urol Oncol 31:211–218CrossRefPubMedCentralPubMedGoogle Scholar
  8. 8.
    Whang YE, Armstrong AJ, Rathmell WK, Godley PA, Kim WY, Pruthi RS, Wallen EM, Crane JM, Moore DT, Grigson G, Morris K, Watkins CP, George DJ (2013) A phase II study of lapatinib, a dual EGFR and HER-2 tyrosine kinase inhibitor, in patients with castration-resistant prostate cancer. Urol Oncol 31:82–86CrossRefPubMedGoogle Scholar
  9. 9.
    Adam RM, Kim J, Lin J, Orsola A, Zhuang L, Rice DC, Freeman MR (2002) Heparin-binding epidermal growth factor-like growth factor stimulates androgen-independent prostate tumor growth and antagonizes androgen receptor function. Endocrinology 143:4599–4608CrossRefPubMedGoogle Scholar
  10. 10.
    Cinar B, De BA, Freeman MR (2005) Post-transcriptional regulation of the androgen receptor by Mammalian target of rapamycin. Cancer Res 65:2547–2553CrossRefPubMedGoogle Scholar
  11. 11.
    Mukhopadhyay NK, Kim J, Cinar B, Ramachandran A, Hager MH, Di VD, Adam RM, Rubin MA, Raychaudhuri P, De BA, Freeman MR (2009) Heterogeneous nuclear ribonucleoprotein K is a novel regulator of androgen receptor translation. Cancer Res 69:2210–2218CrossRefPubMedCentralPubMedGoogle Scholar
  12. 12.
    Lin HK, Wang L, Hu YC, Altuwaijri S, Chang C (2002) Phosphorylation-dependent ubiquitylation and degradation of androgen receptor by Akt require Mdm2 E3 ligase. EMBO J 21:4037–4048CrossRefPubMedCentralPubMedGoogle Scholar
  13. 13.
    Cai C, Portnoy DC, Wang H, Jiang X, Chen S, Balk SP (2009) Androgen receptor expression in prostate cancer cells is suppressed by activation of epidermal growth factor receptor and ErbB2. Cancer Res 69:5202–5209CrossRefPubMedGoogle Scholar
  14. 14.
    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 102:9890–9895CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    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 7:3176–3186CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    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 33:6024–6033CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Zhou H, Mazan-Mamczarz K, Martindale JL, Barker A, Liu Z, Gorospe M, Leedman PJ, Gartenhaus RB, Hamburger AW, Zhang Y (2010) Post-transcriptional regulation of androgen receptor mRNA by an ErbB3 binding protein 1 in prostate cancer. Nucl Acids Res 38:3619–3631CrossRefPubMedCentralPubMedGoogle Scholar
  18. 18.
    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
  19. 19.
    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 396:99–107CrossRefPubMedCentralPubMedGoogle Scholar
  20. 20.
    Ghosh A, Awasthi S, Hamburger AW (2013) ErbB3-binding protein EBP1 decreases ErbB2 levels via a transcriptional mechanism. Oncol Rep 29:1161–1166PubMedCentralPubMedGoogle Scholar
  21. 21.
    Lu Y, Zhou H, Chen W, Zhang Y, Hamburger AW (2011) The ErbB3 binding protein EBP1 regulates ErbB2 protein levels and tamoxifen sensitivity in breast cancer cells. Breast Cancer Res Treat 126:27–36CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Zhang Y, Ali TZ, Zhou H, D’Souza DR, Lu Y, Jaffe J, Liu Z, Passaniti A, Hamburger AW (2010) ErbB3 binding protein 1 represses metastasis-promoting gene anterior gradient protein 2 in prostate cancer. Cancer Res 70:240–248CrossRefPubMedCentralPubMedGoogle Scholar
  23. 23.
    Xia X, Lessor TJ, Zhang Y, Woodford N, Hamburger AW (2001) Analysis of the expression pattern of Ebp1, an ErbB-3-binding protein. Biochem Biophys Res Commun 289:240–244CrossRefPubMedGoogle Scholar
  24. 24.
    Pignon JC, Koopmansch B, Nolens G, Delacroix L, Waltregny D, Winkler R (2009) Androgen receptor controls EGFR and ERBB2 gene expression at different levels in prostate cancer cell lines. Cancer Res 69:2941–2949CrossRefPubMedGoogle Scholar
  25. 25.
    Berger R, Lin DI, Nieto M, Sicinska E, Garraway LA, Adams H, Signoretti S, Hahn WC, Loda M (2006) Androgen-dependent regulation of Her-2/neu in prostate cancer cells. Cancer Res 66:5723–5728CrossRefPubMedGoogle Scholar
  26. 26.
    Chen L, Mooso BA, Jathal MK, Madhav A, Johnson SD, van Spyk E, Mikhailova M, Zierenberg-Ripoll A, Xue L, Vinall RL, deVere White RW, Ghosh PM (2011) Dual EGFR/HER2 inhibition sensitizes prostate cancer cells to androgen withdrawal by suppressing ErbB3. Clin Cancer Res 17:6218–6228CrossRefPubMedCentralPubMedGoogle Scholar
  27. 27.
    Wu HC, Hsieh JT, Gleave ME, Brown NM, Pathak S, Chung LW (1994) Derivation of androgen-independent human LNCaP prostatic cancer cell sublines: role of bone stromal cells. Int J Cancer 57:406–412CrossRefPubMedGoogle Scholar
  28. 28.
    Konecny GE, Pegram MD, Venkatesan N, Finn R, Yang G, Rahmeh M, Untch M, Rusnak DW, Spehar G, Mullin RJ, Keith BR, Gilmer TM, Berger M, Podratz KC, Slamon DJ (2006) Activity of the dual kinase inhibitor lapatinib (GW572016) against HER-2-overexpressing and trastuzumab-treated breast cancer cells. Cancer Res 66:1630–1639CrossRefPubMedGoogle Scholar
  29. 29.
    Wilson TR, Lee DY, Berry L, Shames DS, Settleman J (2011) Neuregulin-1-mediated autocrine signaling underlies sensitivity to HER2 kinase inhibitors in a subset of human cancers. Cancer Cell 20:158–172CrossRefPubMedGoogle Scholar
  30. 30.
    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 265:298–306CrossRefPubMedCentralPubMedGoogle Scholar
  31. 31.
    Dehm SM, Tindall DJ (2006) Molecular regulation of androgen action in prostate cancer. J Cell Biochem 99:333–344CrossRefPubMedGoogle Scholar
  32. 32.
    Agus DB, Sweeney CJ, Morris MJ, Mendelson DS, McNeel DG, Ahmann FR, Wang J, Derynck MK, Ng K, Lyons B, Allison DE, Kattan MW, Scher HI (2007) Efficacy and safety of single-agent pertuzumab (rhuMAb 2C4), a human epidermal growth factor receptor dimerization inhibitor, in castration-resistant prostate cancer after progression from taxane-based therapy. J Clin Oncol 25:675–681CrossRefPubMedGoogle Scholar
  33. 33.
    Zhang Y, Fondell JD, Wang Q, Xia X, Cheng A, Lu ML, Hamburger AW (2002) Repression of androgen receptor mediated transcription by the ErbB-3 binding protein, Ebp1. Oncogene 21:5609–5618CrossRefPubMedGoogle Scholar
  34. 34.
    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 82:683–690CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Tal-Or P, Di Segni A, Lupowitz Z, Pinkas-Kramarski R (2003) Neuregulin promotes autophagic cell death of prostate cancer cells. Prostate 55:147–157CrossRefPubMedGoogle Scholar
  36. 36.
    Wu Z, Gioeli D, Conaway M, Weber MJ, Theodorescu D (2008) Restoration of PTEN expression alters the sensitivity of prostate cancer cells to EGFR inhibitors. Prostate 68:935–944CrossRefPubMedCentralPubMedGoogle Scholar
  37. 37.
    Carver BS, Chapinski C, Wongvipat J, Hieronymus H, Chen Y, Chandarlapaty S, Arora VK, Le C, Koutcher J, Scher H, Scardino PT, Rosen N, Sawyers CL (2011) Reciprocal feedback regulation of PI3 K and androgen receptor signaling in PTEN-deficient prostate cancer. Cancer Cell 19:575–586CrossRefPubMedCentralPubMedGoogle Scholar
  38. 38.
    Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J (2012) Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature 487:505–509CrossRefPubMedCentralPubMedGoogle Scholar
  39. 39.
    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
  40. 40.
    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
  41. 41.
    Zhang Y, Hamburger AW (2005) Specificity and heregulin regulation of Ebp1 (ErbB3 binding protein 1) mediated repression of androgen receptor signalling. Br J Cancer 92:140–146CrossRefPubMedCentralPubMedGoogle Scholar
  42. 42.
    Nethery DE, Ghosh S, Erzurum SC, Kern JA (2007) Inactivation of neuregulin-1 by nitration. Am J Physiol Lung Cell Mol Physiol 292:L287–L293CrossRefPubMedGoogle Scholar
  43. 43.
    Mellinghoff IK, Vivanco I, Kwon A, Tran C, Wongvipat J, Sawyers CL (2004) HER2/neu kinase-dependent modulation of androgen receptor function through effects on DNA binding and stability. Cancer Cell 6:517–527CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Smita Awasthi
    • 1
    • 2
  • Heather Ezelle
    • 1
    • 3
  • Bret A. Hassel
    • 1
    • 3
  • Anne W. Hamburger
    • 1
    • 2
  1. 1.Greenebaum Cancer CenterUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Department of PathologyUniversity of Maryland School of MedicineBaltimoreUSA
  3. 3.Department of Microbiology and ImmunologyUniversity of Maryland School of MedicineBaltimoreUSA

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