Linkage of Progestin and Epidermal Growth Factor Signaling: Phosphorylation of Progesterone Receptors Mediates Transcriptional Hypersensitivity and Increased Ligand-independent Breast Cancer Cell Growth

Overview

Journal Steroids

Publisher Elsevier

Specialty Biochemistry

Date 2006 Dec 19

PMID 17173941

Citations 49

Authors

Andrea R Daniel

Ming Qiu

Emily J Faivre

Julie Hanson Ostrander

Andrew Skildum

Carol A Lange

Affiliations

Soon will be listed here.

Abstract

Progesterone receptor (PR) action is linked to epidermal growth factor (EGF) initiated signaling pathways at multiple levels; mitogen-activated protein kinases (MAPKs) are key mediators of this important cross-talk. Herein, we probed the effects of EGF on PR function and regulation of breast cancer cell growth. EGF stimulated rapid and transient phosphorylation of PR-B Ser294 relative to persistent phosphorylation of this site induced by the synthetic progestin, R5020. EGF induced nuclear translocation and DNA binding of unliganded wild-type, but not mutant PRs containing an Ala at position 294 (S294A). However, EGF alone induced little to no PR-B transcriptional activity; S294A PR-B was transcriptionally impaired. In contrast, pretreatment of cells with EGF (30min) significantly increased the potency and efficacy of wild-type, but not S294A PR transcriptional activity in response to progestin, and enhanced ligand-dependent downregulation of wild-type but not S294A PR. Replacement of Ser294 with aspartic acid (S294D) to mimic phosphorylation at this site decreased receptor stability and, as predicted, heightened progestin-induced transcription relative to wild-type PR-B. RT-PCR demonstrated the Ser294 phosphorylation-dependence of selected PR target genes (TGFalpha and HB-EGF). Surprisingly, PR-B expressing cells growing in soft agar were highly responsive to EGF or progestin, and this was further stimulated by the combination of both hormones. Cells expressing S294A PR exhibited reduced soft agar growth, and were also sensitive to R5020 alone, but failed to respond to EGF. These results suggest that PR Ser294 is an important "sensor" for growth factor inputs that affects PR function and breast cancer cell growth in the absence of progestin or in the presence of low or "sub-threshold" progestin concentrations. PR function likely contributes to breast cancer progression when EGFR family members or their ligands are overexpressed, a condition that predicts low abundance, but highly active and nuclear PR.

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References

Musgrove E, Lee C, Sutherland R . Progestins both stimulate and inhibit breast cancer cell cycle progression while increasing expression of transforming growth factor alpha, epidermal growth factor receptor, c-fos, and c-myc genes. Mol Cell Biol. 1991; 11(10):5032-43. PMC: 361499. DOI: 10.1128/mcb.11.10.5032-5043.1991. View

Qiu M, Olsen A, Faivre E, Horwitz K, Lange C . Mitogen-activated protein kinase regulates nuclear association of human progesterone receptors. Mol Endocrinol. 2003; 17(4):628-42. DOI: 10.1210/me.2002-0378. View

Moore M, Spence J, Kiningham K, Dillon J . Progestin inhibition of cell death in human breast cancer cell lines. J Steroid Biochem Mol Biol. 2006; 98(4-5):218-27. DOI: 10.1016/j.jsbmb.2005.09.008. View

Mansour S, Matten W, Hermann A, Candia J, Rong S, Fukasawa K . Transformation of mammalian cells by constitutively active MAP kinase kinase. Science. 1994; 265(5174):966-70. DOI: 10.1126/science.8052857. View

Clemm D, Sherman L, Boonyaratanakornkit V, Schrader W, Weigel N, Edwards D . Differential hormone-dependent phosphorylation of progesterone receptor A and B forms revealed by a phosphoserine site-specific monoclonal antibody. Mol Endocrinol. 2000; 14(1):52-65. DOI: 10.1210/mend.14.1.0413. View

Woodward T, Xie J, Haslam S . The role of mammary stroma in modulating the proliferative response to ovarian hormones in the normal mammary gland. J Mammary Gland Biol Neoplasia. 2000; 3(2):117-31. DOI: 10.1023/a:1018738721656. View

Rowan B, Weigel N, OMalley B . Phosphorylation of steroid receptor coactivator-1. Identification of the phosphorylation sites and phosphorylation through the mitogen-activated protein kinase pathway. J Biol Chem. 2000; 275(6):4475-83. DOI: 10.1074/jbc.275.6.4475. View

Richer J, Lange C, Manning N, Owen G, Powell R, Horwitz K . Convergence of progesterone with growth factor and cytokine signaling in breast cancer. Progesterone receptors regulate signal transducers and activators of transcription expression and activity. J Biol Chem. 1998; 273(47):31317-26. DOI: 10.1074/jbc.273.47.31317. View

Zhang Z, Funk C, Roy D, Glasser S, Mulholland J . Heparin-binding epidermal growth factor-like growth factor is differentially regulated by progesterone and estradiol in rat uterine epithelial and stromal cells. Endocrinology. 1994; 134(3):1089-94. DOI: 10.1210/endo.134.3.8119147. View

10.

Takimoto G, Hovland A, Tasset D, Melville M, Tung L, Horwitz K . Role of phosphorylation on DNA binding and transcriptional functions of human progesterone receptors. J Biol Chem. 1996; 271(23):13308-16. DOI: 10.1074/jbc.271.23.13308. View

11.

Putti T, Pinder S, Elston C, Lee A, Ellis I . Breast pathology practice: most common problems in a consultation service. Histopathology. 2005; 47(5):445-57. DOI: 10.1111/j.1365-2559.2005.02246.x. View

12.

Skildum A, Faivre E, Lange C . Progesterone receptors induce cell cycle progression via activation of mitogen-activated protein kinases. Mol Endocrinol. 2004; 19(2):327-39. DOI: 10.1210/me.2004-0306. View

13.

Lu N, Cidlowski J . Glucocorticoid receptor isoforms generate transcription specificity. Trends Cell Biol. 2006; 16(6):301-7. DOI: 10.1016/j.tcb.2006.04.005. View

14.

Richer J, Jacobsen B, Manning N, Abel M, Wolf D, Horwitz K . Differential gene regulation by the two progesterone receptor isoforms in human breast cancer cells. J Biol Chem. 2001; 277(7):5209-18. DOI: 10.1074/jbc.M110090200. View

15.

Bamberger A, Bamberger C, Gellersen B, Schulte H . Modulation of AP-1 activity by the human progesterone receptor in endometrial adenocarcinoma cells. Proc Natl Acad Sci U S A. 1996; 93(12):6169-74. PMC: 39208. DOI: 10.1073/pnas.93.12.6169. View

16.

Lange C . Making sense of cross-talk between steroid hormone receptors and intracellular signaling pathways: who will have the last word?. Mol Endocrinol. 2003; 18(2):269-78. DOI: 10.1210/me.2003-0331. View

17.

Cenni , PICARD . Ligand-independent Activation of Steroid Receptors: New Roles for Old Players. Trends Endocrinol Metab. 1999; 10(2):41-46. DOI: 10.1016/s1043-2760(98)00121-0. View

18.

Richer J, Lange C, Wierman A, Brooks K, Tung L, Takimoto G . Progesterone receptor variants found in breast cells repress transcription by wild-type receptors. Breast Cancer Res Treat. 1998; 48(3):231-41. DOI: 10.1023/a:1005941117247. View

19.

JOEL P, Traish A, Lannigan D . Estradiol-induced phosphorylation of serine 118 in the estrogen receptor is independent of p42/p44 mitogen-activated protein kinase. J Biol Chem. 1998; 273(21):13317-23. DOI: 10.1074/jbc.273.21.13317. View

20.

Haslam S, Counterman L, Nummy K . Effects of epidermal growth factor, estrogen, and progestin on DNA synthesis in mammary cells in vivo are determined by the developmental state of the gland. J Cell Physiol. 1993; 155(1):72-8. DOI: 10.1002/jcp.1041550110. View