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EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism

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Abstract

Changes in lamellipod extension and chemotaxis in response to EGF were analysed for MTLn3 cells (a metastatic cell line derived from the 13762NF rat mammary adenocarcinoma). Addition of EGF produced a cessation of ruffling followed by extension of hyaline lamellipods containing increased amounts of F-actin at the growing edge. A non-metastatic cell line (MTC) derived from the same tumor did not show such responses. Lamellipod extension was maximal within 5 min, followed by retraction and resumption of ruffling. Maximal area increases due to lamellipod extension occurred at about 5 nM EGF. Chemotactic and chemokinetic responses, measured using a microchemotaxis chamber, were also greatest at 5 nm. Cytochalasin D inhibited EGF-stimulated responses including lamellipod extension, increases in F-actin in lamellipods, and chemotaxis. Nocodazole affected chemotaxis at higher concentrations but not EGF-induced lamellipod extension. We conclude that polymerization of F-actin at the leading edges of lamellipods is necessary for extension of lamellipods and chemotaxis of MTLn3 cells in response to EGF. The motility and chemotaxis responses of this metastatic cell line have strong similarities to those seen in well-characterized chemotactic cells such as Dictyostelium and neutrophils.

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References

  1. Rosenberg S, 1987, Surgical Treatment of Metastatic Cancer. Lippincott: Philadelphia.

    Google Scholar 

  2. Fidler IJ and Balch CM, 1987, The biology of cancer metastasis and implications for therapy. Curr Prob Surg, 24, 129–209.

    CAS  Google Scholar 

  3. Sugarbaker EV, 1979, Cancer metastasis: a product of tumor-host interactions. Curr Prob Cancer,3, 1–59.

    Article  CAS  Google Scholar 

  4. Nabi IR, Watanabe H and Raz A, 1992, Autocrine motility factor and its receptor: role in cell locomotion and metastasis. Cancer Metastasis Rev, 11, 5–20.

    Article  PubMed  CAS  Google Scholar 

  5. Talmadge JE and Fidler IJ, 1989, Phenotypic heterogeneity and metastasis. In: Sirica AE, ed. The Pathobiology of Neoplasia. New York:Plenum Press, pp. 547–72.

    Google Scholar 

  6. Lu C and Kerbel RS, 1994, Cytokines, growth factors and the loss of negative growth controls in the progression of human cutaneous malignant melanoma. Curr Opin Oncol, 6, 212–20.

    PubMed  CAS  Google Scholar 

  7. Yeatman TJ and Nicolson GL, 1993, Molecular basis of tumor progression: mechanisms of organ-specific tumor metastasis. Seminars Surg Oncol, 9, 256–63.

    CAS  Google Scholar 

  8. Rinker-Schaeffer CW, Partin AW, Isaacs WB, et al. 1994, Molecular and cellular changes associated with the acquisition of metastatic ability by prostatic cancer cells. Prostate, 25, 249–265.

    PubMed  CAS  Google Scholar 

  9. Monsky WL and Chen WT, 1993, Proteases of cell adhesion proteins in cancer. Seminars Cancer Biol, 4, 251–8.

    CAS  Google Scholar 

  10. Aznavoorian S, Murphy AN, Stetler-Stevenson WG, et al. 1993, Molecular aspects of tumor cell invasion and metastasis. Cancer, 71, 1368–83.

    Article  PubMed  CAS  Google Scholar 

  11. Jones JG, Segall J and Condeelis J, 1991, Molecular analysis of amoeboid chemotaxis: parallel observations in amoeboid phagocytes and metastatic tumor cells. Experientia Suppl, 59, 1–16.

    CAS  Google Scholar 

  12. Condeelis J, 1993, Life at the leading edge: the formation of cell protrusion. Ann Rev Cell Biol, 9, 411–44.

    PubMed  CAS  Google Scholar 

  13. Devreotes PN and Zigmond SH, 1988, Chemotaxis in eukaryotic cells: a focus on leukocytes and Dictyostelium. Ann Rev Cell Biol, 4, 649–86.

    PubMed  CAS  Google Scholar 

  14. Silletti S and Raz A, 1993, Autocrine motility factor is a growth factor. Biochem Biophys Res Commun, 194, 446–57.

    Article  PubMed  CAS  Google Scholar 

  15. Stracke M, Liotta LA and Schiffmann E, 1993, The role of autotaxin and other motility stimulating factors in the regulation of tumor cell motility. Symp Soc Exp Biol, 47, 197–214.

    PubMed  CAS  Google Scholar 

  16. Hoelting T, Siperstein AE, Clark OH, et al. 1994, Epidermal growth factor enhances proliferation, migration, and invasion of follicular and papillary thyroid cancer in vitro and in vivo. J Clin Endocr Metab, 79, 401–8.

    Article  PubMed  CAS  Google Scholar 

  17. Pedersen PH, Ness GO, Engebraaten O, et al. 1994, Heterogeneous response to the growth factors [EGF, PDGF (bb), TGF-alpha, bFGF, IL-2] on glioma spheroid growth, migration and invasion. Int J Cancer, 56, 255–61.

    PubMed  CAS  Google Scholar 

  18. Wang X, Kamiyama K, Iguchi I, et al. 1994, Enhancement of fibronectin-induced migration of corneal epithelial cells by cytokines. Invest Ophthalmol Vis Sci, 35, 4001–7.

    PubMed  CAS  Google Scholar 

  19. Royce LS and Baum BJ, 1991, Physiologic levels of salivary epidermal growth factor stimulate migration of an oral epithelial cell line. Biochim Biophys Acta, 1092, 401–3.

    Article  PubMed  CAS  Google Scholar 

  20. Grotendorst GR, Soma Y, Takehara K, et al. 1989, EGF and TGF-alpha are potent chemoattractants for endothelial cells and EGF-like peptides are present at sites of tissue regeneration. J Cell Physiol, 139, 617–23.

    Article  PubMed  CAS  Google Scholar 

  21. Blay J and Brown KD, 1985, Epidermal growth factor promotes the chemotactic migration of cultured rat intestinal epithelial cells. J Cell Physiol, 124, 107–12.

    Article  PubMed  CAS  Google Scholar 

  22. Kundra V, Escobedo JA, Kazlauskas A, et al. 1994, Regulation of chemotaxis by the platelet-derived growth factor receptor-beta. Nature, 367, 474–6.

    Article  PubMed  CAS  Google Scholar 

  23. Yenush L, Kundra V, White MF, et al. 1994, Functional domains of the insulin receptor responsible for chemotactic signaling. J Biol Chem, 269, 100–4.

    PubMed  CAS  Google Scholar 

  24. Li Y, Bhargava MM, Joseph A, et al. 1994, Effect of hepatocyte growth factor/scatter factor and other growth factors on motility and morphology of non-tumorigenic and tumor cells. In Vitro Cell Devel Biol An, 30A, 105–10.

    CAS  Google Scholar 

  25. Gherardi E, Sharpe M, Lane K, et al. 1993, Hepatocyte growth factor/scatter factor (HGF/SF), the c-met receptor and the behaviour of epithelial cells. Symp Soc Exp Biol, 47, 163–81.

    PubMed  CAS  Google Scholar 

  26. Khazaie K, Schirrmacher V and Lichtner RB, 1993, EGF receptor in neoplasia and metastasis. Cancer Metastasis Rev, 12, 255–74.

    Article  PubMed  CAS  Google Scholar 

  27. Weidner N and Gasparini G, 1994, Determination of epidermal growth factor receptor provides additional prognostic information to measuring tumor angiogenesis in breast carcinoma patients. Breast Canc Res Treat, 29, 97–107.

    Article  CAS  Google Scholar 

  28. Fox SB, Smith K, Hollyer J, et al. 1994, The epidermal growth factor receptor as a prognostic marker: results of 370 patients and review of 3009 patients. Breast Canc Res Treat, 29, 41–9.

    Article  CAS  Google Scholar 

  29. Veale D, Kerr N, Gibson GJ, et al. 1993, The relationship of quantitative epidermal growth factor receptor expression in non-small cell lung cancer to long term survival. Br J Cancer, 68, 162–5.

    PubMed  CAS  Google Scholar 

  30. Mayer A, Takimoto M, Fritz E, et al. 1993, The prognostic significance of proliferating cell nuclear antigen, epidermal growth factor receptor, and mdr gene expression in colorectal cancer. Cancer, 71, 2454–60.

    Article  PubMed  CAS  Google Scholar 

  31. Itakura Y, Sasano H, Shiga C, et al. 1994, Epidermal growth factor receptor overexpression in esophageal carcinoma. An immunohistochemical study correlated with clinicopathologic findings and DNA amplification. Cancer, 74, 795–804.

    Article  PubMed  CAS  Google Scholar 

  32. Peppelenbosch MP, Tertoolen LG, Hage WJ, et al. 1993, Epidermal growth factor-induced actin remodeling is regulated by 5-lipoxygenase and cyclooxygenase products. Cell, 74, 565–75.

    Article  PubMed  CAS  Google Scholar 

  33. Kuppuswamy D and Pike LJ, 1991, Desensitization of the EGF receptor alters its ability to undergo EGF-induced dimerization. Cell Signalling, 3, 107–17.

    Article  PubMed  CAS  Google Scholar 

  34. Osherov N and Levitzki A, 1994, Epidermal-growthfactor-dependent activation of the src-family kinases. Eur J Biochem, 225, 1047–53.

    Article  PubMed  CAS  Google Scholar 

  35. Dadabay CY, Patton E, Cooper JA, et al. 1991, Lack of correlation between changes in polyphosphoinositide levels and actin/gelsolin complexes in A431 cells treated with epidermal growth factor. J Cell Biol, 112, 1151–6.

    Article  PubMed  CAS  Google Scholar 

  36. Price JE, Sauder DN and Fidler IJ, 1988, Tumorigenicity and metastatic behavior in nude mice of two human squamous cell carcinoma lines that differ in production of the cytokine ETAF/IL-1. J Invest Dermatol, 91, 258–62.

    Article  PubMed  CAS  Google Scholar 

  37. Neri A, Welch D, Kawaguchi T, et al. 1982, Development and biologic properties of malignant cell sublines and clones of a spontaneously metastasizing rat mammary adenocarcinoma. J Natl Cancer Inst, 68, 507–17.

    PubMed  CAS  Google Scholar 

  38. Welch DR, Neri A and Nicolson GL, 1983, Comparison of ‘spontaneous’ and ‘experimental’ metastasis using rat 13762 mammary adenocarcinoma metastatic cell clones. Invasion Metastasis, 3, 65–80.

    PubMed  CAS  Google Scholar 

  39. Welch DR, Krizman DB and Nicolson GL, 1984, Multiple phenotypic divergence of mammary adenocarcinoma cell lines. I. In vitro and in vivo properties. Clin Exp Metastasis, 2, 333–55.

    Article  PubMed  CAS  Google Scholar 

  40. Kaufmann AM, Khazaie K, Wiedemuth M, et al. 1994, Expression of epidermal growth factor receptor correlates with metastatic potential of 13762NF rat mammary adenocarcinoma cells. Int J Oncol, 4, 1149–55.

    CAS  Google Scholar 

  41. Lichtner RB, Wiedemuth M, Kittmann A, et al. 1992, Ligand-induced activation of epidermal growth factor receptor in intact rat mammary adenocarcinoma cells without detectable receptor phosphorylation. J Biol Chem, 267, 11872–80.

    PubMed  CAS  Google Scholar 

  42. McKenna NM and Wang YL, 1989, Culturing cells on the microscope stage. Meth Cell Biol, 29, 195–205.

    CAS  Google Scholar 

  43. Soil DR, 1988, ‘DMS,’ a computer-assisted system for quantitating motility, the dynamics of cytoplasmic flow, and pseudopod formation: its application to Dictyostelium chemotaxis. Cell Motil Cytoskeleton, 10, 91–106.

    Article  Google Scholar 

  44. Condeelis J and Hall AL, 1991, Measurement of actin polymerization and cross-linking in agonist-stimulated cells. Meth Enzymol, 196, 486–96.

    PubMed  CAS  Google Scholar 

  45. Sampath P and Pollard TD, 1991, Effects of cytochalasin, phalloidin, and pH on the elongation of actin filaments. Biochemistry, 19, 1973–80.

    Article  Google Scholar 

  46. Lichtner RB, Wiedemuth M, Noeske-Jungblut C, et al. 1993, Rapid effects of EGF on cytoskeletal structures and adhesive properties of highly metastatic rat mammary adenocarcinoma cells. Clin Exp Metastasis, 11, 113–25.

    Article  PubMed  CAS  Google Scholar 

  47. Jordan MA, Thrower D and Wilson L, 1992, Effects of vinblastine, podophyllotoxin and nocodazole on mitotic spindles. Implications for the role of microtubule dynamics in mitosis. J Cell Sci, 102, 401–16.

    PubMed  CAS  Google Scholar 

  48. Chinkers M, McKanna JA and Cohen S, 1981, Rapid rounding of human epidermoid carcinoma cells A-431 induced by epidermal growth factor. J Cell Biol, 88, 422–9.

    Article  PubMed  CAS  Google Scholar 

  49. Chinkers M, McKanna JA and Cohen S, 1979, Rapid induction of morphological changes in human carcinoma cells A-431 by epidermal growth factor. J Cell Biol, 83, 260–5.

    Article  PubMed  CAS  Google Scholar 

  50. Miyata Y, Nishida E and Sakai H, 1988, Growth factor- and phorbol ester-induced changes in cell morphology analyzed by digital image processing. Exp Cell Res, 175, 286–97.

    Article  PubMed  CAS  Google Scholar 

  51. van Larebeke NA, Bracke ME and Mareel MM, 1992, Simple method for quantification of fast plasma membrane movements. Cytometry, 13, 1–8.

    Article  PubMed  Google Scholar 

  52. Welsh JB, Gill GN, Rosenfeld MG, et al. 1991, A negative feedback loop attenuates EGF-induced morphological changes. J Cell Biol, 114, 533–43.

    Article  PubMed  CAS  Google Scholar 

  53. Kadowaki T, Koyasu S, Nishida E, et al. 1986, Insulin-like growth factors, insulin, and epidermal growth factor cause rapid cytoskeletal reorganization in KB cells. Clarification of the roles of type I insulin-like growth factor receptors and insulin receptors. J Biol Chem, 261, 16141–7.

    PubMed  CAS  Google Scholar 

  54. Wrann MM and Fox CF, 1979, Identification of epidermal growth factor receptors in a hyperproducing human epidermoid carcinoma cell line. J Biol Chem, 254, 8083–6.

    PubMed  CAS  Google Scholar 

  55. Lichtner RB and Schirrmacher V, 1990, Cellular distribution and biological activity of epidermal growth factor receptors in A431 cells are influenced by cell-cell contact. J Cell Physiol, 144, 303–12.

    Article  PubMed  CAS  Google Scholar 

  56. Koyasu S, Kadowaki T, Nishida E, et al. 1988, Alteration in growth, morphology, and cytoskeletal structures of KB cells induced by epidermal growth factor and transforming growth factor-beta. Exp Cell Res, 176, 107–16.

    Article  PubMed  CAS  Google Scholar 

  57. Gill GN and Lazar CS, 1981, Increased phosphotyrosine content and inhibition of proliferation in EGF-treated A431 cells. Nature, 293, 305–7.

    Article  PubMed  CAS  Google Scholar 

  58. Kawamoto T, Sato JD, Le A, et al. 1983, Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc Natl Acad Sci USA, 80, 1337–41.

    Article  PubMed  CAS  Google Scholar 

  59. Richer M, Castillo MA, Rieber MS, et al. 1988, Decrease in tumor-cell attachment and in a 140-kDa fibronectin receptor correlate with greater expression of multiple 34-kDa surface proteins and cytoplasmic 54-kDa components. Int J Cancer, 41, 96–100.

    Google Scholar 

  60. Nakajima M, Welch DR, Wynn DM, et al. 1993, Serum and plasma M(r) 92,000 progelatinase levels correlate with spontaneous metastasis of rat 13762NF mammary adenocarcinoma. Cancer Res, 53, 5802–7.

    PubMed  CAS  Google Scholar 

  61. Schechter B, Arnon R, Wilchek M, et al. 1991, Indirect immunotargeting of cis-Pt to human epidermoid carcinoma KB using the avidin-biotin system. Int J Cancer, 48, 167–72.

    PubMed  CAS  Google Scholar 

  62. Aboud-Pirak E, Hurwitz E, Pirak ME, et al. 1988, Efficacy of antibodies to epidermal growth factor receptor against KB carcinoma in vitro and in nude mice. J Natl Cancer Inst, 80, 1605–11.

    Article  PubMed  CAS  Google Scholar 

  63. Harvath L, Falk W and Leonard EJ, 1980, Rapid quantitation of neutrophil chemotaxis: use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J Immunol Meth, 37, 39–45.

    Article  CAS  Google Scholar 

  64. Rijken PJ, Hage WJ, van Bergen en Henegouwen PM, et al. 1991, Epidermal growth factor induces rapid reorganization of the actin microfilament system in human A431 cells. J Cell Sci, 100, 491–9.

    PubMed  CAS  Google Scholar 

  65. Schlessinger J and Geiger B, 1981, Epidermal growth factor induces redistribution of actin and alpha-actinin in human epidermal carcinoma cells. Exp Cell Res, 134, 273–9.

    Article  PubMed  CAS  Google Scholar 

  66. Ridley AJ, 1994, Membrane ruffling and signal transduction. Bioessays, 16, 321–7.

    Article  PubMed  CAS  Google Scholar 

  67. Oster G and Perelson A, 1987, The physics of cell motility. J Cell Sci, 8, 35–54.

    CAS  Google Scholar 

  68. Egelhoff TT and Spudich JA, 1991, Molecular genetics of cell migration: Dictyostelium as a model system. Trends Genetics, 7, 161–6.

    CAS  Google Scholar 

  69. Hall AL, Warren V, Dharmawardhane S, et al. 1989, Identification of actin nucleation activity and polymerization inhibitor in ameboid cells: their regulation by chemotactic stimulation. J Cell Biol, 109, 2207–13.

    Article  PubMed  CAS  Google Scholar 

  70. Theriot JA and Mitchison TJ, 1992, Comparison of actin and cell surface dynamics in motile fibroblasts. J Cell Biol, 119, 367–77.

    Article  PubMed  CAS  Google Scholar 

  71. Bates SE, Valverius EM, Ennis BW, et al. 1990, Expression of the transforming growth factor-alpha/epidermal growth factor receptor pathway in normal human breast epithelial cells. Endocrinology, 126, 596–607.

    PubMed  CAS  Google Scholar 

  72. Coleman S, Silberstein GB and Daniel CW, 1988, Ductal morphogenesis in the mouse mammary gland: evidence supporting a role for epidermal growth factor. Develop Biol, 127, 304–15.

    Article  PubMed  CAS  Google Scholar 

  73. Snedeker SM, Brown CF and DiAugustine RP, 1991, Expression and functional properties of transforming growth factor alpha and epidermal growth factor during mouse mammary gland ductal morphogenesis. Proc Natl Acad Sci USA, 88, 276–80.

    Article  PubMed  CAS  Google Scholar 

  74. Atnip KD, Carter LM, Nicolson GL, et al. 1987, Chemotactic response of rat mammary adenocarcinoma cell clones to tumor-derived cytokines. Biochem Biophys Res Commun, 146, 996–1002.

    Article  PubMed  CAS  Google Scholar 

  75. Cunningham CC, 1992, Actin structural proteins in cell motility. Cancer Metastasis Rev, 11, 69–77.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY, USA

    Jeffrey E. Segall, Susan Masseling, Joseph Helft, Amanda Chan & John Condeelis

  2. Department of Radiation Oncology, Albert Einstein College of Medicine, Bronx, NY, USA

    Sangeeta Tyerech & Lucia Boselli

  3. Department of Pathology, Albert Einstein College of Medicine, Bronx, NY, USA

    Joan Jones

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  1. Jeffrey E. Segall
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  2. Sangeeta Tyerech
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Segall, J.E., Tyerech, S., Boselli, L. et al. EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism. Clin Exp Metast 14, 61–72 (1996). https://doi.org/10.1007/BF00157687

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