Fibronectin and type IV collagen activate ERα AF-1 by c-Src pathway: effect on breast cancer cell motility

The expression of estrogen receptor alpha (ERα) is generally associated with a less invasive and aggressive phenotype in breast carcinoma. In an attempt to understand the role of ERα in regulating breast cancer cells invasiveness, we have demonstrated that cell adhesion on fibronectin (Fn) and type IV Collagen (Col) induces ERα-mediated transcription and reduces cell migration in MCF-7 and in MDA-MB-231 cell lines expressing ERα. Analysis of deleted mutants of ERα indicates that the transcriptional activation function (AF)-1 is required for ERα-mediated transcription as well as for the inhibition of cell migration induced by cell adhesion on extracellular matrix (ECM) proteins. In addition, the nuclear localization signal region and some serine residues in the AF-1 of the ERα are both required for the regulation of cell invasiveness as we have observed in HeLa cells. It is worth noting that c-Src activation is coincident with adhesion of cells to ECM proteins and that the inhibition of c-Src activity by PP2 or the expression of a dominant-negative c-Src abolishes ERα-mediated transcription and partially reverts the inhibition of cell invasiveness in ERα-positive cancer cells. These findings address the integrated role of ECM proteins and ERα in influencing breast cancer cell motility through a mechanism that involves c-Src and seems not to be related to a specific cell type.

[1]  D. Lannigan,et al.  Estradiol-induced Phosphorylation of Serine 118 in the Estrogen Receptor Is Independent of p42/p44 Mitogen-activated Protein Kinase* , 1998, The Journal of Biological Chemistry.

[2]  G. Gallick,et al.  Src family kinases in tumor progression and metastasis , 2003, Cancer and Metastasis Reviews.

[3]  E. Howard,et al.  Differential influence of antiestrogens on the in vitro release of gelatinases (type IV collagenases) by invasive and non-invasive breast cancer cells , 1997, Clinical & Experimental Metastasis.

[4]  J. Parsons,et al.  Complex formation with focal adhesion kinase: A mechanism to regulate activity and subcellular localization of Src kinases. , 1999, Molecular biology of the cell.

[5]  C. Lowell,et al.  Deficiency of Src family kinases p59/61hck and p58c-fgr results in defective adhesion-dependent neutrophil functions , 1996, The Journal of cell biology.

[6]  K. Yamada,et al.  Integrins can collaborate with growth factors for phosphorylation of receptor tyrosine kinases and MAP kinase activation: roles of integrin aggregation and occupancy of receptors , 1996, The Journal of cell biology.

[7]  N. Weigel.  Steroid hormone receptors and their regulation by phosphorylation. , 1996, The Biochemical journal.

[8]  J. Thiery,et al.  Phosphorylation of Tyrosine Residues 31 and 118 on Paxillin Regulates Cell Migration through an Association with Crk in Nbt-II Cells , 2000, The Journal of cell biology.

[9]  W. Muller,et al.  Mammary tumors expressing the neu proto-oncogene possess elevated c-Src tyrosine kinase activity , 1994, Molecular and cellular biology.

[10]  D. Vorojeikina,et al.  Phosphorylation of Tyrosine 537 on the Human Estrogen Receptor Is Required for Binding to an Estrogen Response Element (*) , 1995, The Journal of Biological Chemistry.

[11]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[12]  R. Juliano,et al.  Cell Anchorage Permits Efficient Signal Transduction Between Ras and Its Downstream Kinases* , 1997, The Journal of Biological Chemistry.

[13]  J. Brugge,et al.  Integrins and signal transduction pathways: the road taken. , 1995, Science.

[14]  R. Cardiff,et al.  Activation of the c-Src tyrosine kinase is required for the induction of mammary tumors in transgenic mice. , 1994, Genes & development.

[15]  D. Picard,et al.  Activation of the unliganded estrogen receptor by EGF involves the MAP kinase pathway and direct phosphorylation. , 1996, The EMBO journal.

[16]  M. Maa,et al.  Potentiation of epidermal growth factor receptor-mediated oncogenesis by c-Src: implications for the etiology of multiple human cancers. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Daniel Metzger,et al.  Activation of the Estrogen Receptor Through Phosphorylation by Mitogen-Activated Protein Kinase , 1995, Science.

[18]  A. Lucas,et al.  ERβ Inhibits Proliferation and Invasion of Breast Cancer Cells. , 2001, Endocrinology.

[19]  B. Katzenellenbogen,et al.  Estrogen action via the cAMP signaling pathway: stimulation of adenylate cyclase and cAMP-regulated gene transcription. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M Karin,et al.  Potentiation of estrogen receptor activation function 1 (AF-1) by Src/JNK through a serine 118-independent pathway. , 2001, Molecular endocrinology.

[21]  R. Assoian,et al.  Integrin-dependent activation of MAP kinase: a link to shape-dependent cell proliferation. , 1995, Molecular biology of the cell.

[22]  T. Hunter,et al.  Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases , 1996, Molecular and cellular biology.

[23]  H. Rochefort,et al.  Estradiol and fibulin‐1 inhibit motility of human ovarian‐ and breast‐cancer cells induced by fibronectin , 1998, International journal of cancer.

[24]  Robert Clarke,et al.  Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines , 1992, Journal of cellular physiology.

[25]  M E Doerr,et al.  The Roles of Integrins and Extracellular Matrix Proteins in the Insulin-like Growth Factor I-stimulated Chemotaxis of Human Breast Cancer Cells (*) , 1996, The Journal of Biological Chemistry.

[26]  P. Leder,et al.  Signal transduction pathways activated and required for mammary carcinogenesis in response to specific oncogenes , 1998, Oncogene.

[27]  T. Hunter,et al.  Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase , 1994, Nature.

[28]  T. Hunter,et al.  Fibronectin-stimulated signaling from a focal adhesion kinase-c-Src complex: involvement of the Grb2, p130cas, and Nck adaptor proteins , 1997, Molecular and cellular biology.

[29]  K. Korach,et al.  Disruption of the mouse oestrogen receptor gene: resulting phenotypes and experimental findings. , 1995, Biochemical Society transactions.

[30]  L. Sheffield.  C-Src activation by ErbB2 leads to attachment-independent growth of human breast epithelial cells. , 1998, Biochemical and biophysical research communications.

[31]  K. Vuori,et al.  CAS/Crk Coupling Serves as a “Molecular Switch” for Induction of Cell Migration , 1998, The Journal of cell biology.

[32]  Y. Yazaki,et al.  Matrix/Integrin Interaction Activates the Mitogen-activated Protein Kinase, p44 and p42(*) , 1995, The Journal of Biological Chemistry.

[33]  B. O’Malley,et al.  Dopaminergic and ligand-independent activation of steroid hormone receptors. , 1991, Science.

[34]  T. Yeatman,et al.  Role of Src expression and activation in human cancer , 2000, Oncogene.

[35]  D. Crowe,et al.  Transcriptional inhibition of matrix metalloproteinase 9 (MMP-9) activity by a c-fos/estrogen receptor fusion protein is mediated by the proximal AP-1 site of the MMP-9 promoter and correlates with reduced tumor cell invasion. , 1999, Neoplasia.

[36]  P. Chambon,et al.  The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties. , 1989, The EMBO journal.

[37]  J. Guan,et al.  Role of focal adhesion kinase in integrin signaling. , 1997, The international journal of biochemistry & cell biology.

[38]  M. Schwartz,et al.  Growth factor activation of MAP kinase requires cell adhesion , 1997, The EMBO journal.

[39]  D. Chalbos,et al.  Unliganded and liganded estrogen receptors protect against cancer invasion via different mechanisms. , 2000, Molecular endocrinology.

[40]  S. Hanks,et al.  Identification of p130Cas as a Mediator of Focal Adhesion Kinase–promoted Cell Migration , 1998, The Journal of cell biology.

[41]  H. Varmus,et al.  c-Src enhances the spreading of src-/- fibroblasts on fibronectin by a kinase-independent mechanism. , 1995, Genes & development.

[42]  A. Philips,et al.  Synthetic antiestrogens modulate induction of pS2 and cathepsin-D messenger ribonucleic acid by growth factors and adenosine 3',5'-monophosphate in MCF7 cells. , 1993, Endocrinology.