title: The G protein-coupled estrogen receptor-1, GPER-1, promotes fibrillogenesis via a Shc-dependent pathway resulting in anchorage-independent growth.
abstract: The G protein-coupled estrogen receptor-1, GPER-1, coordinates fibronectin (FN) matrix assembly and release of heparan-bound epidermal growth factor (HB-EGF). This mechanism of action results in the recruitment of FN-engaged integrin α5β1 to fibrillar adhesions and the formation of integrin α5β1-Shc adaptor protein complexes. Here, we show that GPER-1 stimulation of murine 4 T1 or human SKBR3 breast cancer cells with 17β-estradiol (E2β) promotes the formation of focal adhesions and actin stress fibers and results in increased cellular adhesion and haptotaxis on FN, but not collagen. These actions are also induced by the xenoestrogen, bisphenol A, and the estrogen receptor (ER) antagonist, ICI 182, 780, but not the inactive stereoisomer, 17α-estradiol (E2α). In addition, we show that GPER-1 stimulation of breast cancer cells allows for FN-dependent, anchorage-independent growth and FN fibril formation in "hanging drop" assays, indicating that these GPER-1-mediated actions occur independently of adhesion to solid substrata. Stable expression of Shc mutant Y317F lacking its primary tyrosyl phosphorylation site disrupts E2β-induced focal adhesion and actin stress fiber formation and abolishes E2β-enhanced haptotaxis on FN and anchorage-dependent growth. Collectively, these data demonstrate that E2β action via GPER-1 enhances cellular adhesivity and FN matrix assembly and allows for anchorage-independent growth, cellular events that may allow for cellular survival, and tumor progression.
authors.affiliation: Division of Hematology and Oncology, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI, 02903, USA.
authors.forename: Hilary T
authors.initials: HT
authors.lastname: Magruder
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Quinn
authors.forename: Jean E
authors.initials: JE
authors.lastname: Schwartzbauer
authors.forename: Jonathan
authors.initials: J
authors.lastname: Reichner
authors.forename: Allan
authors.initials: A
authors.lastname: Huang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Filardo
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001559:Benzhydryl Compounds / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D048708:Cell Growth Processes / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000378:metabolism; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D020847:Estrogen Receptor Modulators / Q000494:pharmacology; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism; D000077267:Fulvestrant; D006801:Humans; D051379:Mice; D009154:Mutation / Q000235:genetics; D010636:Phenols / Q000494:pharmacology; D010766:Phosphorylation / Q000235:genetics; D011960:Receptors, Estrogen / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D055765:Shc Signaling Adaptor Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D022502:Stress Fibers / Q000187:drug effects*; D000098826:Bisphenol A Compounds
chemical_list: D000199:Actins; D001559:Benzhydryl Compounds; D004815:Epidermal Growth Factor; D004958:Estradiol; D020847:Estrogen Receptor Modulators; D005353:Fibronectins; D000077267:Fulvestrant; D010636:Phenols; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D055765:Shc Signaling Adaptor Proteins; C109584:GPER1 protein, human; C532507:GPER1 protein, mouse; C006780:bisphenol A; D000098826:Bisphenol A Compounds
references.citation: Cell. 2007 Dec 14;131(6):1109-23
references.pmid: 18083101
references.citation: Cancer Res. 2008 May 1;68(9):3185-92
references.pmid: 18451144
references.citation: Breast Cancer Res Treat. 2011 Jul;128(2):457-66
references.pmid: 21607586
references.citation: Clin Cancer Res. 2007 Oct 1;13(19):5798-804
references.pmid: 17908971
references.citation: Neoplasia. 2007 Feb;9(2):147-58
references.pmid: 17356711
references.citation: Cancer Res. 1986 May;46(5):2526-31
references.pmid: 2421880
references.citation: Chin J Cancer. 2013 Feb;32(2):75-83
references.pmid: 23237212
references.citation: Am J Surg. 2013 Nov;206(5):698-703
references.pmid: 24011568
references.citation: Environ Pollut. 2011 Jan;159(1):212-218
references.pmid: 20875696
references.citation: J Cell Sci. 1998 Oct;111 ( Pt 19):2933-43
references.pmid: 9730985
references.citation: J Cell Biol. 1998 Nov 2;143(3):861-73
references.pmid: 9813103
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18267-72
references.pmid: 19826086
references.citation: J Cell Biol. 1995 Jul;130(2):441-50
references.pmid: 7542248
references.citation: Mol Endocrinol. 2000 Oct;14(10):1649-60
references.pmid: 11043579
references.citation: J Biol Chem. 1996 Apr 5;271(14):8424-9
references.pmid: 8626541
references.citation: Cell Mol Life Sci. 2013 Nov;70(22):4243-53
references.pmid: 23756580
references.citation: Mol Endocrinol. 2009 Jul;23(7):1052-64
references.pmid: 19342448
references.citation: J Biol Chem. 1997 Jan 17;272(3):1718-24
references.pmid: 8999851
references.citation: Exp Cell Res. 1996 Feb 1;222(2):360-9
references.pmid: 8598224
references.citation: Ann Anat. 1999 May;181(3):231-6
references.pmid: 10363104
references.citation: J Cell Biol. 2007 Oct 8;179(1):23-31
references.pmid: 17908916
references.citation: Cell Calcium. 2006 Mar;39(3):275-92
references.pmid: 16406008
references.citation: Biol Cell. 2000 Oct;92(7):477-94
references.pmid: 11229600
references.citation: Exp Cell Res. 2005 Sep 10;309(1):48-55
references.pmid: 15992798
references.citation: Biochemistry. 2005 Sep 20;44(37):12329-43
references.pmid: 16156646
references.citation: J Biol Chem. 2009 Mar 20;284(12):8148-59
references.pmid: 19144637
references.citation: Blood. 2012 Feb 23;119(8):1946-55
references.pmid: 22096252
references.citation: J Biol Chem. 2000 Nov 24;275(47):36532-40
references.pmid: 10976102
references.citation: Curr Opin Cell Biol. 2001 Oct;13(5):546-54
references.pmid: 11544022
references.citation: J Vis Exp. 2011 May 06;(51):
references.pmid: 21587162
references.citation: Environ Health Perspect. 2012 Aug;120(8):1177-82
references.pmid: 22552965
references.citation: Cell. 1992 Aug 7;70(3):389-99
references.pmid: 1643657
references.citation: Growth Factors. 1990;3(2):97-103
references.pmid: 2206558
references.citation: Immunol Rev. 2003 Feb;191:183-95
references.pmid: 12614360
references.citation: J Mol Endocrinol. 2010 Feb;44(2):115-25
references.pmid: 19850646
references.citation: EMBO J. 1997 Feb 17;16(4):706-16
references.pmid: 9049300
references.citation: J Cell Sci. 2005 Oct 1;118(Pt 19):4427-36
references.pmid: 16159961
references.citation: Oncogene. 2001 Oct 1;20(44):6322-30
references.pmid: 11607835
references.citation: J Cell Biol. 1994 Feb;124(4):619-26
references.pmid: 8106557
references.citation: Breast Cancer Res. 2006;8(6):R73
references.pmid: 17196107
references.citation: Oncogene. 2010 Oct 14;29(41):5559-67
references.pmid: 20676142
references.citation: J Biol Chem. 2000 Apr 7;275(14):10673-82
references.pmid: 10744764
references.citation: J Cell Biol. 1996 Jul;134(2):573-83
references.pmid: 8707839
references.citation: Clin Cancer Res. 2006 Nov 1;12(21):6359-66
references.pmid: 17085646
references.citation: Oncogene. 2010 Oct 14;29(41):5556-8
references.pmid: 20711240
references.citation: J Biol Chem. 2000 Nov 17;275(46):36423-9
references.pmid: 10964917
references.citation: J Thromb Haemost. 2008 Aug;6(8):1253-61
references.pmid: 18513212
references.citation: Cell. 1981 Oct;26(2 Pt 2):259-67
references.pmid: 6174240
references.citation: Cell Growth Differ. 2000 Feb;11(2):123-33
references.pmid: 10714768
references.citation: J Immunol. 2009 Nov 1;183(9):5468-76
references.pmid: 19828641
references.citation: Carcinogenesis. 1998 Mar;19(3):445-55
references.pmid: 9525279
references.citation: PLoS One. 2013 Sep 09;8(9):e72999
references.pmid: 24039841
references.citation: J Cell Biol. 1999 Mar 8;144(5):1019-31
references.pmid: 10085298
references.citation: Angiogenesis. 2009;12(2):165-75
references.pmid: 19219555
references.citation: J Biol Chem. 1998 Aug 14;273(33):21125-31
references.pmid: 9694867
references.citation: Cell. 1998 Sep 4;94(5):625-34
references.pmid: 9741627
references.citation: Breast Cancer Res Treat. 1996;39(1):93-102
references.pmid: 8738609
references.citation: Breast Cancer Res Treat. 2010 Aug;123(1):51-8
references.pmid: 19902352
references.citation: J Biol Chem. 2010 Nov 5;285(45):34875-84
references.pmid: 20739287
references.citation: Curr Opin Cell Biol. 2005 Oct;17(5):509-16
references.pmid: 16099636
references.citation: J Biol Chem. 2002 May 31;277(22):19703-8
references.pmid: 11912200
references.citation: EMBO Rep. 2001 May;2(5):431-7
references.pmid: 11375936
references.citation: J Cell Biol. 1993 Apr;121(1):155-62
references.pmid: 8458867
references.citation: Biol Cell. 2014 Feb;106(2):72-82
references.pmid: 24405025
references.citation: Arterioscler Thromb Vasc Biol. 2002 Jun 1;22(6):927-33
references.pmid: 12067900
references.citation: J Cell Biol. 1996 Aug;134(3):793-9
references.pmid: 8707856
references.citation: J Cell Physiol. 2007 Mar;210(3):750-6
references.pmid: 17187346
references.citation: Endocrinology. 2001 Jul;142(7):3214-22
references.pmid: 11416044
references.citation: Endocrinology. 2006 Mar;147(3):1203-12
references.pmid: 16339197
references.citation: Blood. 1989 Jul;74(1):14-8
references.pmid: 2752106
references.citation: Annu Rev Immunol. 2007;25:619-47
references.pmid: 17201681
references.citation: Mol Biol Cell. 2002 Oct;13(10):3521-31
references.pmid: 12388754
references.citation: Cell. 1996 Nov 15;87(4):733-43
references.pmid: 8929541
references.citation: J Biol Chem. 1994 Oct 14;269(41):25243-6
references.pmid: 7929214
references.citation: BMC Cancer. 2009 Jan 12;9:12
references.pmid: 19138410
references.citation: Mol Endocrinol. 2002 Jan;16(1):70-84
references.pmid: 11773440
references.citation: Cancer Res. 2000 Feb 1;60(3):728-32
references.pmid: 10676660
references.citation: Curr Protoc Stem Cell Biol. 2014 Feb 06;28:2B.6.1-2B.6.23
references.pmid: 24510769