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. 1998 Apr 1;17(7):2008–2018. doi: 10.1093/emboj/17.7.2008

Activation of the Src/p21ras/Erk pathway by progesterone receptor via cross-talk with estrogen receptor.

A Migliaccio 1, D Piccolo 1, G Castoria 1, M Di Domenico 1, A Bilancio 1, M Lombardi 1, W Gong 1, M Beato 1, F Auricchio 1
PMCID: PMC1170546  PMID: 9524123

Abstract

The molecular mechanisms by which ovarian hormones stimulate growth of breast tumors are unclear. It has been reported previously that estrogens activate the signal-transducing Src/p21(ras)/Erk pathway in human breast cancer cells via an interaction of estrogen receptor (ER) with c-Src. We now show that progestins stimulate human breast cancer T47D cell proliferation and induce a similar rapid and transient activation of the pathway which, surprisingly, is blocked not only by anti-progestins but also by anti-estrogens. In Cos-7 cells transfected with the B isoform of progesterone receptor (PRB), progestin activation of the MAP kinase pathway depends on co-transfection of ER. A transcriptionally inactive PRB mutant also activates the signaling pathway, demonstrating that this activity is independent of transcriptional effects. PRB does not interact with c-Src but associates via the N-terminal 168 amino acids with ER. This association is required for the signaling pathway activation by progestins. We propose that ER transmits to the Src/p21(ras)/Erk pathway signals received from the agonist-activated PRB. These findings reveal a hitherto unrecognized cross-talk between ovarian hormones which could be crucial for their growth-promoting effects on cancer cells.

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Selected References

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  1. Akiyama T., Ogawara H. Use and specificity of genistein as inhibitor of protein-tyrosine kinases. Methods Enzymol. 1991;201:362–370. doi: 10.1016/0076-6879(91)01032-w. [DOI] [PubMed] [Google Scholar]
  2. Albanese C., Johnson J., Watanabe G., Eklund N., Vu D., Arnold A., Pestell R. G. Transforming p21ras mutants and c-Ets-2 activate the cyclin D1 promoter through distinguishable regions. J Biol Chem. 1995 Oct 6;270(40):23589–23597. doi: 10.1074/jbc.270.40.23589. [DOI] [PubMed] [Google Scholar]
  3. Altucci L., Addeo R., Cicatiello L., Dauvois S., Parker M. G., Truss M., Beato M., Sica V., Bresciani F., Weisz A. 17beta-Estradiol induces cyclin D1 gene transcription, p36D1-p34cdk4 complex activation and p105Rb phosphorylation during mitogenic stimulation of G(1)-arrested human breast cancer cells. Oncogene. 1996 Jun 6;12(11):2315–2324. [PubMed] [Google Scholar]
  4. Aronica S. M., Kraus W. L., Katzenellenbogen B. S. Estrogen action via the cAMP signaling pathway: stimulation of adenylate cyclase and cAMP-regulated gene transcription. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8517–8521. doi: 10.1073/pnas.91.18.8517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Auricchio F., Di Domenico M., Migliaccio A., Castoria G., Bilancio A. The role of estradiol receptor in the proliferative activity of vanadate on MCF-7 cells. Cell Growth Differ. 1995 Feb;6(2):105–113. [PubMed] [Google Scholar]
  6. Beato M., Herrlich P., Schütz G. Steroid hormone receptors: many actors in search of a plot. Cell. 1995 Dec 15;83(6):851–857. doi: 10.1016/0092-8674(95)90201-5. [DOI] [PubMed] [Google Scholar]
  7. Berthois Y., Katzenellenbogen J. A., Katzenellenbogen B. S. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2496–2500. doi: 10.1073/pnas.83.8.2496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Blackmore P. F., Beebe S. J., Danforth D. R., Alexander N. Progesterone and 17 alpha-hydroxyprogesterone. Novel stimulators of calcium influx in human sperm. J Biol Chem. 1990 Jan 25;265(3):1376–1380. [PubMed] [Google Scholar]
  9. Braunsberg H., Coldham N. G., Leake R. E., Cowan S. K., Wong W. Actions of a progestogen on human breast cancer cells: mechanisms of growth stimulation and inhibition. Eur J Cancer Clin Oncol. 1987 May;23(5):563–571. doi: 10.1016/0277-5379(87)90321-x. [DOI] [PubMed] [Google Scholar]
  10. Chalbos D., Vignon F., Keydar I., Rochefort H. Estrogens stimulate cell proliferation and induce secretory proteins in a human breast cancer cell line (T47D). J Clin Endocrinol Metab. 1982 Aug;55(2):276–283. doi: 10.1210/jcem-55-2-276. [DOI] [PubMed] [Google Scholar]
  11. Clarke C. L., Sutherland R. L. Progestin regulation of cellular proliferation. Endocr Rev. 1990 May;11(2):266–301. doi: 10.1210/edrv-11-2-266. [DOI] [PubMed] [Google Scholar]
  12. Davis R. J. The mitogen-activated protein kinase signal transduction pathway. J Biol Chem. 1993 Jul 15;268(20):14553–14556. [PubMed] [Google Scholar]
  13. Deng T., Karin M. c-Fos transcriptional activity stimulated by H-Ras-activated protein kinase distinct from JNK and ERK. Nature. 1994 Sep 8;371(6493):171–175. doi: 10.1038/371171a0. [DOI] [PubMed] [Google Scholar]
  14. Di Domenico M., Castoria G., Bilancio A., Migliaccio A., Auricchio F. Estradiol activation of human colon carcinoma-derived Caco-2 cell growth. Cancer Res. 1996 Oct 1;56(19):4516–4521. [PubMed] [Google Scholar]
  15. Dickson R. B., Lippman M. E. Growth factors in breast cancer. Endocr Rev. 1995 Oct;16(5):559–589. doi: 10.1210/edrv-16-5-559. [DOI] [PubMed] [Google Scholar]
  16. Dérijard B., Hibi M., Wu I. H., Barrett T., Su B., Deng T., Karin M., Davis R. J. JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell. 1994 Mar 25;76(6):1025–1037. doi: 10.1016/0092-8674(94)90380-8. [DOI] [PubMed] [Google Scholar]
  17. Endoh H., Sasaki H., Maruyama K., Takeyama K., Waga I., Shimizu T., Kato S., Kawashima H. Rapid activation of MAP kinase by estrogen in the bone cell line. Biochem Biophys Res Commun. 1997 Jun 9;235(1):99–102. doi: 10.1006/bbrc.1997.6746. [DOI] [PubMed] [Google Scholar]
  18. Fantl V., Stamp G., Andrews A., Rosewell I., Dickson C. Mice lacking cyclin D1 are small and show defects in eye and mammary gland development. Genes Dev. 1995 Oct 1;9(19):2364–2372. doi: 10.1101/gad.9.19.2364. [DOI] [PubMed] [Google Scholar]
  19. Filmus J., Robles A. I., Shi W., Wong M. J., Colombo L. L., Conti C. J. Induction of cyclin D1 overexpression by activated ras. Oncogene. 1994 Dec;9(12):3627–3633. [PubMed] [Google Scholar]
  20. Gong W., Chávez S., Beato M. Point mutation in the ligand-binding domain of the progesterone receptor generates a transdominant negative phenotype. Mol Endocrinol. 1997 Sep;11(10):1476–1485. doi: 10.1210/mend.11.10.9991. [DOI] [PubMed] [Google Scholar]
  21. Guy C. T., Muthuswamy S. K., Cardiff R. D., Soriano P., Muller W. J. Activation of the c-Src tyrosine kinase is required for the induction of mammary tumors in transgenic mice. Genes Dev. 1994 Jan;8(1):23–32. doi: 10.1101/gad.8.1.23. [DOI] [PubMed] [Google Scholar]
  22. Hanke J. H., Gardner J. P., Dow R. L., Changelian P. S., Brissette W. H., Weringer E. J., Pollok B. A., Connelly P. A. Discovery of a novel, potent, and Src family-selective tyrosine kinase inhibitor. Study of Lck- and FynT-dependent T cell activation. J Biol Chem. 1996 Jan 12;271(2):695–701. doi: 10.1074/jbc.271.2.695. [DOI] [PubMed] [Google Scholar]
  23. Herber B., Truss M., Beato M., Müller R. Inducible regulatory elements in the human cyclin D1 promoter. Oncogene. 1994 Apr;9(4):1295–1304. [PubMed] [Google Scholar]
  24. Hissom J. R., Moore M. R. Progestin effects on growth in the human breast cancer cell line T-47D--possible therapeutic implications. Biochem Biophys Res Commun. 1987 Jun 15;145(2):706–711. doi: 10.1016/0006-291x(87)91022-9. [DOI] [PubMed] [Google Scholar]
  25. Humphreys R. C., Lydon J., O'Malley B. W., Rosen J. M. Mammary gland development is mediated by both stromal and epithelial progesterone receptors. Mol Endocrinol. 1997 Jun;11(6):801–811. doi: 10.1210/mend.11.6.9891. [DOI] [PubMed] [Google Scholar]
  26. Ignar-Trowbridge D. M., Nelson K. G., Bidwell M. C., Curtis S. W., Washburn T. F., McLachlan J. A., Korach K. S. Coupling of dual signaling pathways: epidermal growth factor action involves the estrogen receptor. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4658–4662. doi: 10.1073/pnas.89.10.4658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kastner P., Krust A., Turcotte B., Stropp U., Tora L., Gronemeyer H., Chambon P. Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. EMBO J. 1990 May;9(5):1603–1614. doi: 10.1002/j.1460-2075.1990.tb08280.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kato S., Endoh H., Masuhiro Y., Kitamoto T., Uchiyama S., Sasaki H., Masushige S., Gotoh Y., Nishida E., Kawashima H. Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase. Science. 1995 Dec 1;270(5241):1491–1494. doi: 10.1126/science.270.5241.1491. [DOI] [PubMed] [Google Scholar]
  29. Kerkhoff E., Rapp U. R. Induction of cell proliferation in quiescent NIH 3T3 cells by oncogenic c-Raf-1. Mol Cell Biol. 1997 May;17(5):2576–2586. doi: 10.1128/mcb.17.5.2576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kiss R., Paridaens R. J., Heuson J. C., Danguy A. J. Effect of progesterone on cell proliferation in the MXT mouse hormone-sensitive mammary neoplasm. J Natl Cancer Inst. 1986 Jul;77(1):173–178. [PubMed] [Google Scholar]
  31. Korach K. S. Insights from the study of animals lacking functional estrogen receptor. Science. 1994 Dec 2;266(5190):1524–1527. doi: 10.1126/science.7985022. [DOI] [PubMed] [Google Scholar]
  32. Kumar V., Green S., Stack G., Berry M., Jin J. R., Chambon P. Functional domains of the human estrogen receptor. Cell. 1987 Dec 24;51(6):941–951. doi: 10.1016/0092-8674(87)90581-2. [DOI] [PubMed] [Google Scholar]
  33. Lavoie J. N., L'Allemain G., Brunet A., Müller R., Pouysségur J. Cyclin D1 expression is regulated positively by the p42/p44MAPK and negatively by the p38/HOGMAPK pathway. J Biol Chem. 1996 Aug 23;271(34):20608–20616. doi: 10.1074/jbc.271.34.20608. [DOI] [PubMed] [Google Scholar]
  34. Lieberherr M., Grosse B., Duchambon P., Drüeke T. A functional cell surface type receptor is required for the early action of 1,25-dihydroxyvitamin D3 on the phosphoinositide metabolism in rat enterocytes. J Biol Chem. 1989 Dec 5;264(34):20403–20406. [PubMed] [Google Scholar]
  35. Lippman M. E., Dickson R. B. Mechanisms of growth control in normal and malignant breast epithelium. Recent Prog Horm Res. 1989;45:383–440. doi: 10.1016/b978-0-12-571145-6.50012-1. [DOI] [PubMed] [Google Scholar]
  36. Longman S. M., Buehring G. C. Oral contraceptives and breast cancer. In vitro effect of contraceptive steroids on human mammary cell growth. Cancer. 1987 Jan 15;59(2):281–287. doi: 10.1002/1097-0142(19870115)59:2<281::aid-cncr2820590218>3.0.co;2-a. [DOI] [PubMed] [Google Scholar]
  37. Lovec H., Sewing A., Lucibello F. C., Müller R., Möröy T. Oncogenic activity of cyclin D1 revealed through cooperation with Ha-ras: link between cell cycle control and malignant transformation. Oncogene. 1994 Jan;9(1):323–326. [PubMed] [Google Scholar]
  38. Lubahn D. B., Moyer J. S., Golding T. S., Couse J. F., Korach K. S., Smithies O. Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11162–11166. doi: 10.1073/pnas.90.23.11162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lukas J., Bartkova J., Bartek J. Convergence of mitogenic signalling cascades from diverse classes of receptors at the cyclin D-cyclin-dependent kinase-pRb-controlled G1 checkpoint. Mol Cell Biol. 1996 Dec;16(12):6917–6925. doi: 10.1128/mcb.16.12.6917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lydon J. P., DeMayo F. J., Funk C. R., Mani S. K., Hughes A. R., Montgomery C. A., Jr, Shyamala G., Conneely O. M., O'Malley B. W. Mice lacking progesterone receptor exhibit pleiotropic reproductive abnormalities. Genes Dev. 1995 Sep 15;9(18):2266–2278. doi: 10.1101/gad.9.18.2266. [DOI] [PubMed] [Google Scholar]
  41. Manni A., Badger B., Wright C., Ahmed S. R., Demers L. M. Effects of progestins on growth of experimental breast cancer in culture: interaction with estradiol and prolactin and involvement of the polyamine pathway. Cancer Res. 1987 Jun 15;47(12):3066–3071. [PubMed] [Google Scholar]
  42. Mendoza C., Soler A., Tesarik J. Nongenomic steroid action: independent targeting of a plasma membrane calcium channel and a tyrosine kinase. Biochem Biophys Res Commun. 1995 May 16;210(2):518–523. doi: 10.1006/bbrc.1995.1690. [DOI] [PubMed] [Google Scholar]
  43. Migliaccio A., Di Domenico M., Castoria G., de Falco A., Bontempo P., Nola E., Auricchio F. Tyrosine kinase/p21ras/MAP-kinase pathway activation by estradiol-receptor complex in MCF-7 cells. EMBO J. 1996 Mar 15;15(6):1292–1300. [PMC free article] [PubMed] [Google Scholar]
  44. Musgrove E. A., Hamilton J. A., Lee C. S., Sweeney K. J., Watts C. K., Sutherland R. L. Growth factor, steroid, and steroid antagonist regulation of cyclin gene expression associated with changes in T-47D human breast cancer cell cycle progression. Mol Cell Biol. 1993 Jun;13(6):3577–3587. doi: 10.1128/mcb.13.6.3577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Musgrove E. A., Lee C. S., Buckley M. F., Sutherland R. L. Cyclin D1 induction in breast cancer cells shortens G1 and is sufficient for cells arrested in G1 to complete the cell cycle. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8022–8026. doi: 10.1073/pnas.91.17.8022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Musgrove E. A., Lilischkis R., Cornish A. L., Lee C. S., Setlur V., Seshadri R., Sutherland R. L. Expression of the cyclin-dependent kinase inhibitors p16INK4, p15INK4B and p21WAF1/CIP1 in human breast cancer. Int J Cancer. 1995 Nov 15;63(4):584–591. doi: 10.1002/ijc.2910630420. [DOI] [PubMed] [Google Scholar]
  47. Muslin A. J., MacNicol A. M., Williams L. T. Raf-1 protein kinase is important for progesterone-induced Xenopus oocyte maturation and acts downstream of mos. Mol Cell Biol. 1993 Jul;13(7):4197–4202. doi: 10.1128/mcb.13.7.4197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Pappas T. C., Gametchu B., Watson C. S. Membrane estrogen receptors identified by multiple antibody labeling and impeded-ligand binding. FASEB J. 1995 Mar;9(5):404–410. doi: 10.1096/fasebj.9.5.7896011. [DOI] [PubMed] [Google Scholar]
  49. Pietras R. J., Arboleda J., Reese D. M., Wongvipat N., Pegram M. D., Ramos L., Gorman C. M., Parker M. G., Sliwkowski M. X., Slamon D. J. HER-2 tyrosine kinase pathway targets estrogen receptor and promotes hormone-independent growth in human breast cancer cells. Oncogene. 1995 Jun 15;10(12):2435–2446. [PubMed] [Google Scholar]
  50. Planas-Silva M. D., Weinberg R. A. Estrogen-dependent cyclin E-cdk2 activation through p21 redistribution. Mol Cell Biol. 1997 Jul;17(7):4059–4069. doi: 10.1128/mcb.17.7.4059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Power R. F., Mani S. K., Codina J., Conneely O. M., O'Malley B. W. Dopaminergic and ligand-independent activation of steroid hormone receptors. Science. 1991 Dec 13;254(5038):1636–1639. doi: 10.1126/science.1749936. [DOI] [PubMed] [Google Scholar]
  52. Prall O. W., Sarcevic B., Musgrove E. A., Watts C. K., Sutherland R. L. Estrogen-induced activation of Cdk4 and Cdk2 during G1-S phase progression is accompanied by increased cyclin D1 expression and decreased cyclin-dependent kinase inhibitor association with cyclin E-Cdk2. J Biol Chem. 1997 Apr 18;272(16):10882–10894. doi: 10.1074/jbc.272.16.10882. [DOI] [PubMed] [Google Scholar]
  53. Russmann C., Truss M., Fix A., Naumer C., Herrmann T., Schmitt J., Stollhof J., Beigang R., Beato M. Crosslinking of progesterone receptor to DNA using tuneable nanosecond, picosecond and femtosecond UV laser pulses. Nucleic Acids Res. 1997 Jun 15;25(12):2478–2484. doi: 10.1093/nar/25.12.2478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Sagata N., Daar I., Oskarsson M., Showalter S. D., Vande Woude G. F. The product of the mos proto-oncogene as a candidate "initiator" for oocyte maturation. Science. 1989 Aug 11;245(4918):643–646. doi: 10.1126/science.2474853. [DOI] [PubMed] [Google Scholar]
  55. Sartorius C. A., Groshong S. D., Miller L. A., Powell R. L., Tung L., Takimoto G. S., Horwitz K. B. New T47D breast cancer cell lines for the independent study of progesterone B- and A-receptors: only antiprogestin-occupied B-receptors are switched to transcriptional agonists by cAMP. Cancer Res. 1994 Jul 15;54(14):3868–3877. [PubMed] [Google Scholar]
  56. Schlessinger J. How receptor tyrosine kinases activate Ras. Trends Biochem Sci. 1993 Aug;18(8):273–275. doi: 10.1016/0968-0004(93)90031-h. [DOI] [PubMed] [Google Scholar]
  57. Schlessinger J., Ullrich A. Growth factor signaling by receptor tyrosine kinases. Neuron. 1992 Sep;9(3):383–391. doi: 10.1016/0896-6273(92)90177-f. [DOI] [PubMed] [Google Scholar]
  58. Sicinski P., Donaher J. L., Parker S. B., Li T., Fazeli A., Gardner H., Haslam S. Z., Bronson R. T., Elledge S. J., Weinberg R. A. Cyclin D1 provides a link between development and oncogenesis in the retina and breast. Cell. 1995 Aug 25;82(4):621–630. doi: 10.1016/0092-8674(95)90034-9. [DOI] [PubMed] [Google Scholar]
  59. Tora L., Mullick A., Metzger D., Ponglikitmongkol M., Park I., Chambon P. The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties. EMBO J. 1989 Jul;8(7):1981–1986. doi: 10.1002/j.1460-2075.1989.tb03604.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Tora L., White J., Brou C., Tasset D., Webster N., Scheer E., Chambon P. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell. 1989 Nov 3;59(3):477–487. doi: 10.1016/0092-8674(89)90031-7. [DOI] [PubMed] [Google Scholar]
  61. Vignon F., Bouton M. M., Rochefort H. Antiestrogens inhibit the mitogenic effect of growth factors on breast cancer cells in the total absence of estrogens. Biochem Biophys Res Commun. 1987 Aug 14;146(3):1502–1508. doi: 10.1016/0006-291x(87)90819-9. [DOI] [PubMed] [Google Scholar]
  62. Warner M. R. Effect of perinatal oestrogen on the pretreatment required for mouse mammary lobular formation in vitro. J Endocrinol. 1978 Apr;77(1):1–10. doi: 10.1677/joe.0.0770001. [DOI] [PubMed] [Google Scholar]
  63. Watters J. J., Campbell J. S., Cunningham M. J., Krebs E. G., Dorsa D. M. Rapid membrane effects of steroids in neuroblastoma cells: effects of estrogen on mitogen activated protein kinase signalling cascade and c-fos immediate early gene transcription. Endocrinology. 1997 Sep;138(9):4030–4033. doi: 10.1210/endo.138.9.5489. [DOI] [PubMed] [Google Scholar]
  64. Weisz A., Bresciani F. Estrogen regulation of proto-oncogenes coding for nuclear proteins. Crit Rev Oncog. 1993;4(4):361–388. [PubMed] [Google Scholar]
  65. Zhang Y., Bai W., Allgood V. E., Weigel N. L. Multiple signaling pathways activate the chicken progesterone receptor. Mol Endocrinol. 1994 May;8(5):577–584. doi: 10.1210/mend.8.5.8058067. [DOI] [PubMed] [Google Scholar]

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