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Prolactin modulates phosphorylation, signaling and trafficking of epidermal growth factor receptor in human T47D breast cancer cells

Oncogene volume 25, pages 7565–7576 (2006)Cite this article

Abstract

Prolactin (PRL) is a polypeptide hormone produced by the anterior pituitary gland and other sites that acts both systemically and locally to cause lactation and other biological effects by interacting with the PRL receptor, a Janus kinase (JAK)2-coupled cytokine receptor family member, and activating downstream signal pathways. Recent evidence suggests PRL is a player in the pathogenesis and progression of breast cancer. Epidermal growth factor (EGF) also has effects on breast tissue, working through its receptors, epidermal growth factor receptor (EGFR) and ErbB-2 (c-neu, HER2), both intrinsic tyrosine kinase growth factor receptors. EGFR promotes pubertal breast ductal morphogenesis in mice, and both EGFR and ErbB-2 are relevant in pathogenesis and behavior of breast and other human cancers. Previous studies showed that PRL and EGF synergize to enhance motility in the human breast cancer cell line, T47D. In this study, we explored crosstalk between the PRL and EGF signaling pathways in T47D cells, with an ultimate aim of understanding how these two important factors might work together in vivo to affect breast cancer behavior. Both PRL and EGF caused robust signaling in T47D cells; PRL acutely activated JAK2, signal transducer and activator of transcription-5 (STAT5), and extracellular signal-regulated kinase-1 and -2 (ERK1 and ERK2), whereas EGF caused EGFR activation and consequent src homology collagen (SHC) activation and ERK activation. Notably, PRL also caused phosphorylation of the EGFR and ErbB-2 at sites detected by PTP101, an antibody that recognizes threonine phosphorylation at consensus motifs for ERK-induced phosphorylation. PRL-induced PTP101-reactive phosphorylation was prevented by pretreatment with PD98059, an ERK pathway inhibitor. Furthermore, PRL synergized with EGF in activating SHC and ERK and transactivating a luciferase reporter driven by c-fos gene enhancer elements, suggesting that PRL allowed markedly enhanced EGF signaling. This was accompanied by substantial inhibition of EGF-induced EGFR downregulation when PRL and EGF cotreatment was compared to EGF treatment alone. This effect of PRL was abrogated by ERK pathway inhibitor pretreatment. Our data suggest that PRL synergistically augments EGF signaling in T47D breast cancer cells at least in part by lessening EGF-induced EGFR downregulation and that this effect requires PRL-induced ERK activity and threonine phosphorylation of EGFR.

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References

  • Acosta JJ, Munoz RM, Gonzalez L, Subtil-Rodriguez A, Dominguez-Caceres MA, Garcia-Martinez JM et al. (2003). Mol Endocrinol 17: 2268–2282.

  • Alessi DT, Cuenda A, Cohen P, Dudley DT, Saltiel A . (1995). J Biol Chem 270: 27489–27494.

  • Alroy I, Yarden Y . (1997). FEBS Lett 410: 83–86.

  • Anderson E, Ferguson JE, Morten H, Shalet SM, Robinson EL, Howell A . (1993). Eur J Cancer 29A: 209–217.

  • Argetsinger LS, Campbell GS, Yang X, Witthuhn BA, Silvennoinen O, Ihle JN et al. (1993). Cell 74: 237–244.

  • Badache A, Hynes NE . (2001). Cancer Res 61: 383–391.

  • Bagowski CP, Stein-Gerlach M, Choidas A, Ullrich A . (1999). EMBO J 18: 5567–5576.

  • Bao J, Alroy I, Waterman H, Schejter ED, Brodie C, Gruenberg J et al. (2000). J Biol Chem 275: 26178–26186.

  • Bargmann CI, Hung MC, Weinberg RA . (1986). Nature 319: 226–230.

  • Bazan JF . (1990). Proc Natl Acad Sci USA 87: 6934–6938.

  • Ben-Jonathan N, Liby K, McFarland M, Zinger M . (2002). Trends Endocrinol Metab 13: 245–250.

  • Ben-Jonathan N, Mershon JL, Allen DL, Steinmetz RW . (1996). Endocr Rev 17: 639–669.

  • Berger MS, Locher GW, Saurer S, Gullick WJ, Waterfield MD, Groner B et al. (1988). Cancer Res 48: 1238–1243.

  • Biscardi JS, Ishizawar RC, Silva CM, Parsons SJ . (2000). Breast Cancer Res 2: 203–210.

  • Bole-Feysot C, Goffin V, Edery M, Binart N, Kelly PA . (1998). Endocrinol Rev 19: 225–268.

  • Bonneterre J, Mauriac L, Weber B, Roche H, Fargeot P, Tubiana-Hulin M et al. (1988). Eur J Cancer Clin Oncol 24: 1851–1853.

  • Boonstra J, Rijken P, Humbel B, Cremers F, Verkleij A, van Bergen en Henegouwen P . (1995). Cell Biol Int 19: 413–430.

  • Burke P, Schooler K, Wiley HS . (2001). Mol Biol Cell 12: 1897–1910.

  • Campbell GS, Argetsinger LS, Ihle JN, Kelly PA, Rillema JA, Carter-Su C . (1994). Proc Natl Acad Sci USA 91: 5232–5236.

  • Ceresa BP, Schmid SL . (2000). Curr Opin Cell Biol 12: 204–210.

  • Chen NY, Holle L, Li W, Peirce SK, Beck MT, Chen WY . (2002). Int J Oncol 20: 813–818.

  • Chen WS, Lazar CS, Poenie M, Tsien RY, Gill GN, Rosenfeld MG . (1987). Nature 328: 820–823.

  • Chughtai N, Schimchowitsch S, Lebrun JJ, Ali S . (2002). J Biol Chem 277: 31107–31114.

  • Clevenger CV, Furth PA, Hankinson SE, Schuler LA . (2003). Endocrinol Rev 24: 1–27.

  • Clevenger CV, Kline JB . (2001). Lupus 10: 706–718.

  • Das R, Vonderhaar BK . (1996a). Breast Cancer Res Treat 40: 141–149.

  • Das R, Vonderhaar BK . (1996b). Oncogene 13: 1139–1145.

  • Davis RJ, Czech MP . (1985). Proc Natl Acad Sci USA 82: 1974–1978.

  • Di Fiore PP, De Camilli P . (2001). Cell 106: 1–4.

  • Dudley DT, Pang L, Decker SJ, Bridges A, Saltiel AR . (1995). Proc Natl Acad Sci USA 92: 7686–7689.

  • Eisenberg A, Biener E, Charlier M, Krishnan RV, Djiane J, Herman B et al. (2004). FEBS Lett 565: 139–142.

  • Favata MF, Horiuchi KY, Manos EJ, Daulerio AJ, Stradley DA, Feeser WS et al. (1998). J Biol Chem 273: 18623–18632.

  • Fenton SE, Sheffield LG . (1993). Mol Biol Cell 4: 773–780.

  • Fenton SE, Sheffield LG . (1997). Exp Cell Res 236: 285–293.

  • Frank SJ, Yi W, Zhao Y, Goldsmith JF, Gilliland G, Jiang J et al. (1995). J Biol Chem 270: 14776–14785.

  • Freeman ME, Kanyicska B, Lerant A, Nagy G . (2000). Physiol Rev 80: 1523–1631.

  • Gibson S, Tu S, Oyer R, Anderson SM, Johnson GL . (1999). J Biol Chem 274: 17612–17618.

  • Ginsburg E, Vonderhaar BK . (1995). Cancer Res 55: 2591–2595.

  • Goffin V, Bernichtein S, Touraine P, Kelly PA . (2005). Endocrinol Rev 26: 400–422.

  • Goffin V, Binart N, Touraine P, Kelly PA . (2002). Annu Rev Physiol 64: 47–67.

  • Goffin V, Touraine P, Pichard C, Bernichtein S, Kelly PA . (1999). Mol Cell Endocrinol 151: 79–87.

  • Graus-Porta D, Beerli RR, Daly JM, Hynes NE . (1997). EMBO J 16: 1647–1655.

  • Grimm SL, Seagroves TN, Kabotyanski EB, Hovey RC, Vonderhaar BK, Lydon JP et al. (2002). Mol Endocrinol 16: 2675–2691.

  • Gutzman JH, Nikolai SE, Rugowski DE, Watters JJ, Schuler LA . (2005). Mol Endocrinol 19: 1765–1778.

  • Gutzman JH, Rugowski DE, Schroeder MD, Watters JJ, Schuler LA . (2004). Mol Endocrinol 18: 3064–3075.

  • Hankinson SE, Willett WC, Michaud DS, Manson JE, Colditz GA, Longcope C et al. (1999). J Natl Cancer Inst 91: 629–634.

  • Haraguchi S, Good RA, Engelman RW, Greene S, Day NK . (1997). Mol Cell Endocrinol 129: 145–155.

  • Harari P . (2004). Endocrinol Relat Cancer 11: 689–708.

  • Heisermann GJ, Gill GN . (1988). J Biol Chem 263: 13152–13158.

  • Holbro T, Civenni G, Hynes NE . (2003). Exp Cell Res 284: 99–110.

  • Horseman ND . (1999). J Mammary Gland Biol Neoplasia 4: 79–88.

  • Hovey RC, Trott JF, Ginsburg E, Goldhar A, Sasaki MM, Fountain SJ et al. (2001). Dev Dyn 222: 192–205.

  • Huang Y, Chang Y, Wang X, Jiang J, Frank SJ . (2004a). Endocrinology 145: 3297–3306.

  • Huang Y, Kim SO, Jiang J, Frank SJ . (2003). J Biol Chem 278: 18902–18913.

  • Huang Y, Kim S-O, Yang N, Jiang J, Frank SJ . (2004b). Mol Endocrinol 18: 1471–1485.

  • Hunter T, Ling N, Cooper JA . (1984). Nature 311: 480–483.

  • Jiang J, Liang L, Kim SO, Zhang Y, Mandler R, Frank SJ . (1998). Biochem Biophys Res Commun 253: 774–779.

  • Johnson JL, Fenton S, Sheffield LG . (1996). J Biol Chem 271: 21574–21578.

  • Karunagaran D, Tzahar E, Beerli RR, Chen X, Graus-Porta D, Ratzkin BJ . (1996). EMBO J 15: 254–264.

  • Kassenbrock CK, Hunter S, Garl P, Johnson GL, Anderson SM . (2002). J Biol Chem 277: 24967–24975.

  • Kim SO, Houtman JC, Jiang J, Ruppert JM, Bertics PJ, Frank SJ . (1999). J Biol Chem 274: 36015–36024.

  • Kim SO, Jiang J, Yi W, Feng GS, Frank SJ . (1998). J Biol Chem 273: 2344–2354.

  • Kim SO, Loesch K, Wang X, Jiang J, Mei L, Cunnick JM . (2002). Endocrinology 143: 4856–4867.

  • Kraus MH, Issing W, Miki T, Popescu NC, Aaronson SA . (1989). Proc Natl Acad Sci USA 86: 9193–9197.

  • Laskin JJ, Sandler AB . (2004). Cancer Treat Rev 30: 1–17.

  • Li X, Huang Y, Jiang J, Frank SJ . (2005). Endocrine Society Program and Abstracts, 87th Annual Meeting. Endocrine Society Press: San Diego, CA, pp 3–91, abstract.

    Google Scholar 

  • Lichtner RB . (2003). Biomed Pharmacother 57: 447–451.

  • Lichtner RB, Menrad A, Sommer A, Klar U, Schneider MR . (2001). Cancer Res 61: 5790–5795.

  • Lin CR, Chen WS, Kruiger W, Stolarsky LS, Weber W, Evans RM et al. (1984). Science 224: 843–848.

  • Liu X, Robinson GW, Wagner KU, Garrett L, Wynshaw-Boris A, Hennighausen L . (1997). Genes Dev 11: 179–186.

  • Llovera M, Pichard C, Bernichtein S, Jeay S, Touraine P, Kelly PA et al. (2000a). Oncogene 19: 4695–4705.

  • Llovera M, Touraine P, Kelly PA, Goffin V . (2000b). Exp Gerontol 35: 41–51.

  • Maus MV, Reilly SC, Clevenger CV . (1999). Endocrinology 140: 5447–5450.

  • McMurray RW, Weidensaul D, Allen SH, Walker SE . (1995). J Rheumatol 22: 2084–2091.

  • Mendelsohn J, Baselga J . (2000). Oncogene 19: 6550–6565.

  • Mendelsohn J, Baselga J . (2003). J Clin Oncol 21: 2787–2799.

  • Nicholson RI, Gee JM, Harper ME . (2001). Eur J Cancer 37: S9–S15.

  • Northwood IC, Gonzalez FA, Wartmann M, Raden DL, Davis RJ . (1991). J Biol Chem 266: 15266–15276.

  • Plowman GD, Green JM, Culouscou JM, Carlton GW, Rothwell VM, Buckley S . (1993). Nature 366: 473–475.

  • Qiu Y, Ravi L, Kung HJ . (1998). Nature 393: 83–85.

  • Quijano Jr VJ, Sheffield LG . (1998). J Biol Chem 273: 1200–1207.

  • Reynolds C, Montone KT, Powell CM, Tomaszewski JE, Clevenger CV . (1997). Endocrinology 138: 5555–5560.

  • Rui H . (2000). In: Oppenheim JJ, Feldman M (eds). Cytokine Reference On-Line, vol. website:www.academicpress.com/cytokinereference,24-h free access. Academic Press, Harcourt: London, UK, pp 267–283.

    Google Scholar 

  • Rui H, Kirken RA, Farrar WL . (1994). J Biol Chem 269: 5364–5368.

  • Schroeder MD, Symowicz J, Schuler LA . (2002). Mol Endocrinol 16: 45–57.

  • Shilo BZ . (2003). Exp Cell Res 284: 140–149.

  • Stern DF . (2000). Breast Cancer Res 2: 176–183.

  • Takishima K, Friedman B, Fujiki H, Rosner MR . (1988). Biochem Biophys Res Commun 157: 740–746.

  • Takishima K, Griswold-Prenner I, Ingebritsen T, Rosner MR . (1991). Proc Natl Acad Sci USA 88: 2520–2524.

  • Toullec D, Pianetti P, Coste H, Bellevergue P, Grand-Perret T, Ajakane M et al. (1991). J Biol Chem 266: 15771–15781.

  • Touraine P, Martini JF, Zafrani B, Durand JC, Labaille F, Malet C et al. (1998). J Clin Endocrinol Metab 83: 667–674.

  • Vonderhaar BK . (1999). Endocr Relat Cancer 6: 389–404.

  • Wennbo H, Kindblom J, Isaksson OG, Tornell J . (1997). Endocrinology 138: 4410–4415.

  • Wiepz GJ, Houtman JC, Cha D, Bertics PJ . (1997). J Cell Physiol 173: 44–53.

  • Wiley HS . (2003). Exp Cell Res 284: 78–88.

  • Wiley HS, Shvartsman SY, Lauffenburger DA . (2003). Trends Cell Biol 13: 43–50.

  • Xie W, Paterson AJ, Chin E, Nabell LM, Kudlow JE . (1997). Mol Endocrinol 11: 1766–1781.

  • Yamauchi T, Ueki K, Tobe K, Tamemoto H, Sekine N, Wada M et al. (1997). Nature 390: 91–96.

  • Yamauchi T, Ueki K, Tobe K, Tamemoto H, Sekine N, Wada M et al. (1998). Endocrinol J 45: 27–31.

  • Yamauchi T, Yamauchi N, Ueki K, Sugiyama T, Waki H, Miki H et al. (2000). J Biol Chem 275: 33937–33944.

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Acknowledgements

We appreciate helpful conversations with Drs X Wang, N Yang, K Loesch, J Cowan, L Deng and K He. This work was supported by NIH Grant DK46395 (to SJF). Parts of this work were presented at the 86th Annual Endocrine Society Meeting in New Orleans, LA, USA, 2004.

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Authors and Affiliations

  1. Department of Medicine, Division of Endocrinology, Diabetes, and Metabolism, University of Alabama at Birmingham, Birmingham, AL, USA

    Y Huang, J Jiang & S J Frank

  2. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    X Li

  3. Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    S J Frank

  4. Endocrinology Section, Medical Service, Veterans Affairs Medical Center, Birmingham, AL, USA

    S J Frank

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Correspondence to S J Frank.

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Huang, Y., Li, X., Jiang, J. et al. Prolactin modulates phosphorylation, signaling and trafficking of epidermal growth factor receptor in human T47D breast cancer cells. Oncogene 25, 7565–7576 (2006). https://doi.org/10.1038/sj.onc.1209740

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