Signaling through ShcA Is Required for Transforming Growth Factor β- and Neu/ErbB-2-Induced Breast Cancer Cell Motility and Invasion

ABSTRACT Cooperation between the Neu/ErbB-2 and transforming growth factor β (TGF-β) signaling pathways enhances the invasive and metastatic capabilities of breast cancer cells; however, the underlying mechanisms mediating this synergy have yet to be fully explained. We demonstrate that TGF-β induces the migration and invasion of mammary tumor explants expressing an activated Neu/ErbB-2 receptor, which requires signaling from autophosphorylation sites located in the C terminus. A systematic analysis of mammary tumor explants expressing Neu/ErbB-2 add-back receptors that couple to distinct signaling molecules has mapped the synergistic effect of TGF-β-induced motility and invasion to signals emanating from tyrosine residues 1226/1227 and 1253 of Neu/ErbB-2. Given that the ShcA adaptor protein is known to interact with Neu/ErbB-2 through these residues, we investigated the importance of this signaling molecule in TGF-β-induced cell motility and invasion. The reduction of ShcA expression rendered cells expressing activated Neu/ErbB-2, or add-back receptors signaling specifically through tyrosines 1226/1227 or 1253, unresponsive to TGF-β-induced motility and invasion. In addition, a dominant-negative form of ShcA, lacking its three known tyrosine phosphorylation sites, completely abrogates the TGF-β-induced migration and invasion of breast cancer cells expressing activated Neu/ErbB-2. Our results implicate signaling through the ShcA adaptor as a key component in the synergistic interaction between these pathways.

[1]  R. Cardiff,et al.  Distinct ErbB-2 coupled signaling pathways promote mammary tumors with unique pathologic and transcriptional profiles. , 2007, Cancer research.

[2]  C. Hudis.  Trastuzumab--mechanism of action and use in clinical practice. , 2007, The New England journal of medicine.

[3]  C. Sotiriou,et al.  Taking gene-expression profiling to the clinic: when will molecular signatures become relevant to patient care? , 2007, Nature Reviews Cancer.

[4]  T. Pawson,et al.  Screening for PTB Domain Binding Partners and Ligand Specificity Using Proteome-Derived NPXY Peptide Arrays , 2007, Molecular and Cellular Biology.

[5]  Robert D. Cardiff,et al.  Insights from transgenic mouse models of ERBB2-induced breast cancer , 2007, Nature Reviews Cancer.

[6]  R. Caprioli,et al.  Multivariable Difference Gel Electrophoresis and Mass Spectrometry , 2007, Molecular & Cellular Proteomics.

[7]  T. Pawson,et al.  Screening for PTB Domain Binding Partners and LigandSpecificity Using Proteome-Derived NPXY Peptide Arrays , 2006, Molecular and Cellular Biology.

[8]  Frederick Y. Wu,et al.  HER2/Neu (ErbB2) signaling to Rac1-Pak1 is temporally and spatially modulated by transforming growth factor beta. , 2006, Cancer research.

[9]  David R Critchley,et al.  The structure and regulation of vinculin. , 2006, Trends in cell biology.

[10]  Brian Bierie,et al.  Tumour microenvironment: TGFβ: the molecular Jekyll and Hyde of cancer , 2006, Nature Reviews Cancer.

[11]  R. Liddington,et al.  Role of vinculin in regulating focal adhesion turnover. , 2006, European journal of cell biology.

[12]  M. Barcellos-Hoff,et al.  Activated type I TGFβ receptor kinase enhances the survival of mammary epithelial cells and accelerates tumor progression , 2006, Oncogene.

[13]  Gavin MacBeath,et al.  A quantitative protein interaction network for the ErbB receptors using protein microarrays , 2006, Nature.

[14]  Greg Yothers,et al.  Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. , 2005, The New England journal of medicine.

[15]  H. Beug,et al.  Molecular requirements for epithelial-mesenchymal transition during tumor progression. , 2005, Current opinion in cell biology.

[16]  Andrés J. García,et al.  Cell adhesion strengthening: contributions of adhesive area, integrin binding, and focal adhesion assembly. , 2005, Molecular biology of the cell.

[17]  C. Heldin,et al.  Non-Smad TGF-β signals , 2005, Journal of Cell Science.

[18]  M. Mann,et al.  Phosphotyrosine interactome of the ErbB-receptor kinase family , 2005, Molecular systems biology.

[19]  Morag Park,et al.  CrkI and CrkII Function as Key Signaling Integrators for Migration and Invasion of Cancer Cells , 2005, Molecular Cancer Research.

[20]  C. Arteaga,et al.  Overexpression of HER2 (erbB2) in Human Breast Epithelial Cells Unmasks Transforming Growth Factor β-induced Cell Motility* , 2004, Journal of Biological Chemistry.

[21]  L. Addadi,et al.  Hierarchical assembly of cell-matrix adhesion complexes. , 2004, Biochemical Society transactions.

[22]  N. Hynes,et al.  Memo mediates ErbB2-driven cell motility , 2004, Nature Cell Biology.

[23]  N. Carragher,et al.  Focal adhesion and actin dynamics: a place where kinases and proteases meet to promote invasion. , 2004, Trends in cell biology.

[24]  Yosef Yarden,et al.  Signal transduction and oncogenesis by ErbB/HER receptors. , 2004, International journal of radiation oncology, biology, physics.

[25]  Joshua LaBaer,et al.  Cooperation of the ErbB2 receptor and transforming growth factor beta in induction of migration and invasion in mammary epithelial cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Martin Bastmeyer,et al.  Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion , 2004, Journal of Cell Science.

[27]  Dana M. Brantley-Sieders,et al.  Increased Malignancy of Neu-Induced Mammary Tumors Overexpressing Active Transforming Growth Factor β1 , 2003, Molecular and Cellular Biology.

[28]  J. Massagué,et al.  Cytostatic and apoptotic actions of TGF-β in homeostasis and cancer , 2003, Nature Reviews Cancer.

[29]  Philip M. Long,et al.  Breast cancer classification and prognosis based on gene expression profiles from a population-based study , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[30]  F. Maurer,et al.  The ErbB2/ErbB3 heterodimer functions as an oncogenic unit: ErbB2 requires ErbB3 to drive breast tumor cell proliferation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[31]  R. Tibshirani,et al.  Repeated observation of breast tumor subtypes in independent gene expression data sets , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[32]  R. Cardiff,et al.  Transforming growth factor β signaling impairs Neu-induced mammary tumorigenesis while promoting pulmonary metastasis , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Massagué,et al.  Mechanisms of TGF-β Signaling from Cell Membrane to the Nucleus , 2003, Cell.

[34]  C. Arteaga,et al.  p38 mitogen-activated protein kinase is required for TGFbeta-mediated fibroblastic transdifferentiation and cell migration. , 2002, Journal of cell science.

[35]  L. Wakefield,et al.  Lifetime exposure to a soluble TGF-beta antagonist protects mice against metastasis without adverse side effects. , 2002, The Journal of clinical investigation.

[36]  W. Muller,et al.  Multiple ErbB-2/Neu Phosphorylation Sites Mediate Transformation through Distinct Effector Proteins* , 2001, The Journal of Biological Chemistry.

[37]  R. Tibshirani,et al.  Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Beningo,et al.  Nascent Focal Adhesions Are Responsible for the Generation of Strong Propulsive Forces in Migrating Fibroblasts , 2001, The Journal of cell biology.

[39]  L. Addadi,et al.  Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates , 2001, Nature Cell Biology.

[40]  M. Moran,et al.  Grb2 and Shc Adapter Proteins Play Distinct Roles in Neu (ErbB-2)-Induced Mammary Tumorigenesis: Implications for Human Breast Cancer , 2001, Molecular and Cellular Biology.

[41]  S. Sadekova,et al.  Distinct tyrosine autophosphorylation sites mediate induction of epithelial mesenchymal like transition by an activated ErbB-2/Neu receptor , 2001, Oncogene.

[42]  H. Moses,et al.  Phosphatidylinositol 3-Kinase Function Is Required for Transforming Growth Factor β-mediated Epithelial to Mesenchymal Transition and Cell Migration* , 2000, The Journal of Biological Chemistry.

[43]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[44]  D. Cheresh,et al.  Bifurcation of Cell Migratory and Proliferative Signaling by the Adaptor Protein Shc , 1999, The Journal of cell biology.

[45]  T. Pawson,et al.  Accelerated Mammary Tumor Development in Mutant Polyomavirus Middle T Transgenic Mice Expressing Elevated Levels of Either the Shc or Grb2 Adapter Protein , 1999, Molecular and Cellular Biology.

[46]  R. Cardiff,et al.  Elevated expression of activated forms of Neu/ErbB‐2 and ErbB‐3 are involved in the induction of mammary tumors in transgenic mice: implications for human breast cancer , 1999, The EMBO journal.

[47]  E. Adamson,et al.  Rescue of the mutant phenotype by reexpression of full-length vinculin in null F9 cells; effects on cell locomotion by domain deleted vinculin. , 1998, Journal of cell science.

[48]  John N. Hutchinson,et al.  Requirement for Both Shc and Phosphatidylinositol 3′ Kinase Signaling Pathways in Polyomavirus Middle T-Mediated Mammary Tumorigenesis , 1998, Molecular and Cellular Biology.

[49]  S. Bull,et al.  neu/erbB-2 amplification identifies a poor-prognosis group of women with node-negative breast cancer. Toronto Breast Cancer Study Group. , 1998, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[50]  E. Adamson,et al.  Vinculin knockout results in heart and brain defects during embryonic development. , 1998, Development.

[51]  M. Moran,et al.  Distinct tyrosine autophosphorylation sites negatively and positively modulate neu-mediated transformation , 1997, Molecular and cellular biology.

[52]  M. Shibuya,et al.  Tyrosine phosphorylation sites at amino acids 239 and 240 of Shc are involved in epidermal growth factor-induced mitogenic signaling that is distinct from Ras/mitogen-activated protein kinase activation , 1997, Molecular and cellular biology.

[53]  Tony Pawson,et al.  The Shc adaptor protein is highly phosphorylated at conserved, twin tyrosine residues (Y239/240) that mediate protein–protein interactions , 1996, Current Biology.

[54]  R. Ezzell,et al.  Targeted disruption of vinculin genes in F9 and embryonic stem cells changes cell morphology, adhesion, and locomotion. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[55]  R. Derynck,et al.  TGF-beta induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors , 1994, The Journal of cell biology.

[56]  W. Muller,et al.  Novel activating mutations in the neu proto-oncogene involved in induction of mammary tumors. , 1994, Molecular and Cellular Biology.

[57]  T Pawson,et al.  Formation of Shc-Grb2 complexes is necessary to induce neoplastic transformation by overexpression of Shc proteins. , 1994, Oncogene.

[58]  R. Cardiff,et al.  Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[59]  R. Cardiff,et al.  Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease , 1992, Molecular and cellular biology.

[60]  S. Aaronson,et al.  The carboxy-terminal domains of erbB-2 and epidermal growth factor receptor exert different regulatory effects on intrinsic receptor tyrosine kinase function and transforming activity. , 1990, Molecular and cellular biology.

[61]  P. Jolicoeur,et al.  Stochastic appearance of mammary tumors in transgenic mice carrying the MMTV/c-neu oncogene , 1989, Cell.

[62]  W Godolphin,et al.  Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. , 1989, Science.

[63]  Cori Bargmann,et al.  Oncogenic activation of the neu‐encoded receptor protein by point mutation and deletion. , 1988, The EMBO journal.

[64]  P. Leder,et al.  Single-step induction of mammary adenocarcinoma in transgenic mice bearing the activated c-neu oncogene , 1988, Cell.

[65]  Cori Bargmann,et al.  Multiple independent activations of the neu oncogene by a point mutation altering the transmembrane domain of p185 , 1986, Cell.

[66]  C. Hoge,et al.  THE NEW ENGLAND JOURNAL OF MEDICINE , 1977, The Lancet.

[67]  H. Smith,et al.  Epithelial cell cultures from normal glandular tissue of mice. , 1974, Journal of the National Cancer Institute.

[68]  R. Caprioli,et al.  Multivariable Difference Gel Electrophoresis and Mass Spectrometry A CASE STUDY ON TRANSFORMING GROWTH FACTOR- (cid:1) AND ERBB2 SIGNALING* □ S , 2006 .

[69]  Y. Shyr,et al.  Transforming growth factor beta-regulated gene expression in a mouse mammary gland epithelial cell line , 2003, Breast Cancer Research.

[70]  J. Wrana,et al.  TGF-beta and the Smad signal transduction pathway. , 2002, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[71]  H. Moses,et al.  Transforming growth factor-beta1 mediates epithelial to mesenchymal transdifferentiation through a RhoA-dependent mechanism. , 2001, Molecular biology of the cell.

[72]  P. Ravdin.  neu/erbB-2 amplification identifies a poor-prognosis group of women with node-negative breast cancer , 1999 .

[73]  B. Geiger,et al.  Overexpression of vinculin suppresses cell motility in BALB/c 3T3 cells. , 1992, Cell motility and the cytoskeleton.

[74]  A. Ullrich,et al.  Identification of autophosphorylation sites of HER2/neu. , 1990, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[75]  M. Sporn,et al.  Transforming growth factor beta. , 1988, Advances in cancer research.

[76]  W. McGuire,et al.  Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. , 1987, Science.

[77]  Frederick Y. Wu,et al.  Transforming Growth Factor (cid:2) (TGF- (cid:2) )-Smad Target Gene Protein Tyrosine Phosphatase Receptor Type Kappa Is Required for TGF- (cid:2) Function† , 2022 .