Induction of cancer cell migration by epidermal growth factor is initiated by specific phosphorylation of tyrosine 1248 of c‐erbB‐2 receptor via epidermal growth factor receptor

Induction of tumor cell migration is a key step in invasion and metastasis. Here we report that the epidermal growth factor (EGF)‐induced cell migration of breast cancer cells is attributed to a transient, rather than a sustained, activation of phospholipase C (PLC) ‐γ1 due to c‐erbB‐2 signaling. EGF stimulation of EGF receptor (EGFR) overexpressing cells resulted in long‐term PLC‐γ1 tyrosine phosphorylation and sustained levels of inositol‐1,4,5‐triphosphate (IP3) and diacylglycerol (DAG) producing sinusoidal calcium oscillations. In contrast, c‐erbB‐2/EGFR expressing cells displayed baseline transient calcium oscillations after EGF treatment due to short‐term PLC‐γ1 tyrosine phosphorylation and short‐term IP3 and DAG turnover. A third cell line expressing a point‐mutated c‐erbB‐2 receptor that lacks the autophosphorylation Y1248 was generated to investigate whether the different PLC‐γ1 activation was attributed to this structure. Neither PLC‐γ1 tyrosine phosphorylation nor IP3 and DAG turnover and calcium oscillations were observed in this cell line, indicating the modulation of the PLC‐γ1 activation time course by c‐erbB‐2 signaling. Induction of cell migration was solely observable in the c‐erbB‐2‐positive cell line as proved by the mode of actin reorganization and a cell migration assay, using a 3D‐ collagen lattice. In summary, c‐erbB‐2 up‐regulation switches on the cell migration program by modulating the time course of PLC‐γ1 activation.

[1]  N. Hynes,et al.  ErbB‐2, the preferred heterodimerization partner of all ErbB receptors, is a mediator of lateral signaling , 1997, The EMBO journal.

[2]  J. Feldner,et al.  Cancer cell motility--on the road from c-erbB-2 receptor steered signaling to actin reorganization. , 2002, Experimental cell research.

[3]  K. Zänker,et al.  Locomotion of tumor cells: a molecular comparison to migrating pre- and postmitotic leukocytes , 2000, Journal of Cancer Research and Clinical Oncology.

[4]  T. Gudermann,et al.  Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol , 1999, Nature.

[5]  S. Rhee,et al.  Regulation of Phosphoinositide-specific Phospholipase C Isozymes* , 1997, The Journal of Biological Chemistry.

[6]  P. Friedl,et al.  Lymphocyte locomotion in three-dimensional collagen gels. Comparison of three quantitative methods for analysing cell trajectories. , 1993, Journal of immunological methods.

[7]  Monilola A. Olayioye,et al.  The ErbB signaling network: receptor heterodimerization in development and cancer , 2000, The EMBO journal.

[8]  F. Maxfield,et al.  Transient increases in cytosolic free calcium appear to be required for the migration of adherent human neutrophils [published erratum appears in J Cell Biol 1990 Mar;110(3):861] , 1990, The Journal of cell biology.

[9]  M. Sliwkowski,et al.  Binding specificities and affinities of egf domains for ErbB receptors , 1999, FEBS letters.

[10]  Careen K. Tang,et al.  EGF Family Receptors and Their Ligands in Human Cancer , 1998 .

[11]  Y. Yarden,et al.  ErbB‐2 is a common auxiliary subunit of NDF and EGF receptors: implications for breast cancer. , 1996, The EMBO journal.

[12]  Y. Yarden,et al.  Neu differentiation factor activation of ErbB-3 and ErbB-4 is cell specific and displays a differential requirement for ErbB-2 , 1995, Molecular and cellular biology.

[13]  G. Assmann,et al.  c‐erbB‐2/EGFR as dominant heterodimerization partners determine a motogenic phenotype in human breast cancer cells , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

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

[15]  J. Putney,et al.  Spatial and temporal aspects of cellular calcium signaling , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  D. Amin,et al.  Relative activities of rat and dog platelet phospholipase A2 and diglyceride lipase. Selective inhibition of diglyceride lipase by RHC 80267. , 1982, The Journal of biological chemistry.

[17]  D. Kennerly,et al.  An indirect pathway of receptor-mediated 1,2-diacylglycerol formation in mast cells. I. IgE receptor-mediated activation of phospholipase D. , 1990, Journal of Immunology.

[18]  M. Peppelenbosch,et al.  Epidermal growth factor activates calcium channels by phospholipase A 2 5 -lipoxygenase-mediated leukotriene C4 production , 1992, Cell.

[19]  F. Oesch,et al.  Prognostic significance of c-erB-2 mRNA in ovarian carcinoma. , 1996, Gynecologic oncology.

[20]  M. Greene,et al.  Structural analysis of p185c-neu and epidermal growth factor receptor tyrosine kinases: oligomerization of kinase domains. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[21]  D. Stern,et al.  Activation state-specific monoclonal antibody detects tyrosine phosphorylated p185neu/erbB-2 in a subset of human breast tumors overexpressing this receptor. , 1995, Cancer research.

[22]  Y. Yarden,et al.  The ErbB signaling network in embryogenesis and oncogenesis: signal diversification through combinatorial ligand‐receptor interactions , 1997, FEBS letters.

[23]  A. Wells,et al.  A Role for Phospholipase C-γ-mediated Signaling in Tumor Cell Invasion , 1999 .

[24]  W. Dougall,et al.  Heterodimerization of epidermal growth factor receptor and wild-type or kinase-deficient Neu: a mechanism of interreceptor kinase activation and transphosphorylation. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Tomohiko Maehama,et al.  The Tumor Suppressor, PTEN/MMAC1, Dephosphorylates the Lipid Second Messenger, Phosphatidylinositol 3,4,5-Trisphosphate* , 1998, The Journal of Biological Chemistry.

[26]  A. Wells,et al.  A role for phospholipase C-gamma-mediated signaling in tumor cell invasion. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[27]  J. Kinet,et al.  PtdIns-3,4,5-P3 A Regulatory Nexus between Tyrosine Kinases and Sustained Calcium Signals , 1998, Cell.

[28]  N. Hynes,et al.  Single-chain antibody-mediated intracellular retention of ErbB-2 impairs Neu differentiation factor and epidermal growth factor signaling , 1995, Molecular and cellular biology.

[29]  P. Seeburg,et al.  Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. , 1985, Science.

[30]  N. Hynes,et al.  The biology of erbB-2/neu/HER-2 and its role in cancer. , 1994, Biochimica et biophysica acta.

[31]  M. Berridge,et al.  The versatility and universality of calcium signalling , 2000, Nature Reviews Molecular Cell Biology.