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Expression and ligand binding of α2β1 integrin on breast carcinoma cells

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We examined the expression and ligand specificity of the α2β1 integrin on human mammary epithelial cells (HMEC) and a panel of breast carcinoma cell lines in vitro. We found that the α2β1 integrin was universally, but quite variably expressed on these cells by FACS analysis. No significant correlation was observed between its expression and other known cellular phenotypes. Substrate attachment assays using blocking antibodies demonstrated that α2β1 integrin served as a receptor for collagen on HMEC and almost all breast carcinoma cells. However, its contribution to laminin binding of these cells appeared to be related to cellular differentiation as evaluated by sex steroid receptor status and by markers of epithelial-mesenchymal transition, i.e. loss of E-cadherin and expression of vimentin. Two different populations of non-malignant immortalized HMEC (184A1N4 and MCF-10A) contained cells capable of using α2β1 integrin as a laminin receptor. Breast cancer cell lines positive for estrogen receptor (ER) and E-cadherin (MCF-7, T47D, ZR75-1) could also use α2β1 integrin as a laminin receptor. Conversely, α2β1 integrin appeared to be incapable of binding to laminin or to be a very minor receptor for laminin on metastatic ER-negative breast carcinoma cells that expressed vimentin (MDA-MB 231, MDA-MB 435, and MDA-MB 436). These findings suggest that the ligand specificity of α2β1 integrin, i.e. its function as a laminin receptor, may be regulated during the malignant progression of breast carcinoma cells. A reduced contribution of α2β1 integrin to the cellular laminin binding appears to be associated with an increased malignant phenotype and with an epithelial-mesenchymal transition of breast carcinoma cells.

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References

  1. Albelda, SM and Buck, CA, 1990, Integrins and other cell adhesion molecules. FASEB J, 4, 2868–80.

    Google Scholar 

  2. Ruoslahti, E, 1991, Integrins. J Clin Invest, 87, 1–5.

    Google Scholar 

  3. Hynes, RO, 1992, Integrins: Versatility, modulation, and signaling in cell adhesion. Cell, 69, 11–25.

    Google Scholar 

  4. Hynes, RO, 1987, Integrins: A family of cell surface receptor. Cell, 48, 549–54.

    Google Scholar 

  5. Blood, CH and Zetter, BR, 1990, Tumor interactions with the vasculature: angiogenesis and tumor metastasis. Biochim Biophys Acta, 1032, 89–118.

    Google Scholar 

  6. Liotta, LA, Steeg, PS and Stetler-Stevenson, WG, 1991, Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell, 64, 327–36.

    Google Scholar 

  7. Auerbach, R, 1988, Patterns of tumor metastasis: organ selectivity in the spread of cancer cells. Lab Invest, 58, 361–4.

    Google Scholar 

  8. Maemura, M and Dickson, RB, 1994, Are cellular adhesion molecules involved in the metastasis of breast cancer? Breast Cancer Res Treat, 32, 239–60.

    Google Scholar 

  9. Albelda, SM, 1993, Role of integrins and other cell adhesion molecules in tumor progression and metastasis. Lab Invest, 68, 4–17.

    Google Scholar 

  10. Pignatelli, M, Smith, MEF and Bodmer, WF, 1990, Low expression of collagen receptors in moderate and poorly differentiated colorectal adenocarcinomas. Br J Cancer, 61, 636–8.

    Google Scholar 

  11. Hall, PA, Coates, P, Lemoine, NR and Horton, MA, 1991, Characterization of integrin chains in normal and neoplastic human pancreas. J Pathol, 165, 33–41.

    Google Scholar 

  12. Korhonen, M, Laitinen, L, Ylänne, J et al., 1992, Integrin distribution in renal cell carcinomas of various grades of malignancy. Am J Pathol, 141, 1161–71.

    Google Scholar 

  13. Suzuki, S, Takahashi, T, Nakamura, S et al., 1993, Alteration of integrin expression in human lung cancer. Jpn J Cancer Res, 84, 168–74.

    Google Scholar 

  14. Jones, JL, Critchley, DR and Walker, RA, 1992, Alteration of stromal protein and integrin expression in breast — a marker of premalignant change? J Pathol, 167, 399–406.

    Google Scholar 

  15. Plantefaber, LC and Hynes, RO, 1989, Changes in integrin receptors on oncogenically transformed cells. Cell, 56, 281–90.

    Google Scholar 

  16. Lin, C-S, Zhang, K and Kramer, R, 1993, α6 integrin is up-regulated in step increments accompanying neoplastic transformation and tumorigenic conversion of human fibroblasts. Cancer Res, 53, 2950–3.

    Google Scholar 

  17. Akiyama, SK, Larjava, H and Yamada, KM, 1990, Differences in the biosynthesis and localization of the fibronectin receptor in normal and transformed cultured human cells. Cancer Res, 50, 1601–7.

    Google Scholar 

  18. Dedhar, S and Saulnier, R, 1990, Alterations in integrin receptor expression on chemically transformed human cells: specific enhancement of laminin and collagen receptor complexes. J Cell Biol, 110, 481–9.

    Google Scholar 

  19. Danen, EHJ, van Muijen, GNP, van de Wiel-van Kemenade, E et al., 1993, Regulation of integrin-mediated adhesion to laminin and collagen in human melanocytes and in non-metastatic and highly metastatic human melanoma cells. Int J Cancer, 54, 315–21.

    Google Scholar 

  20. Dedhar, S, Saulnier, R, Nagle, R, and Overall, CM, 1993, Specific alterations in the expression of α3β1 and α6β4 integrins in highly invasive and metastatic variants of human prostate carcinoma cells selected by in vitro invasion through reconstituted basement membrane. Clin Exp Metastasis, 11, 391–400.

    Google Scholar 

  21. Yamada, KM, Kennedy, DW, Yamada, SS et al., 1990, Monoclonal antibody and synthetic peptide inhibitors of human tumor cell migration. Cancer Res, 50, 4485–96.

    Google Scholar 

  22. Fujita, S, Suzuki, H, Kinoshita, M and Hirohashi, S, 1992, Inhibition of cell attachment, invasion and metastasis of human carcinoma cells by anti-integrin β1, subunit antibody. Jpn J Cancer Res, 83, 1317–26.

    Google Scholar 

  23. Bonkhoff, H, Stein, U and Remberger, K, 1993, Differential expression of α6 and α2 very late antigen integrins in the normal, hyperplastic, and neoplastic prostate: simultaneous demonstration of cell surface receptors and their extracellular ligands. Hum Pathol, 24, 243–8.

    Google Scholar 

  24. Nigam, AK, Savage, FJ, Boulos, PB et al., 1993, Loss of cell-cell and cell-matrix adhesion molecules in colorectal cancer. Br J Cancer, 68, 507–14.

    Google Scholar 

  25. Chen, FA, Repasky, EA and Bankert, RB, 1991, Human lung tumor-associated antigen identified as an extracellular matrix adhesion molecule. J Exp Med, 173, 1111–19.

    Google Scholar 

  26. Zutter, MM, Mazoujian, G and Santoro, SA, 1990, Decreased expression of integrin adhesive protein receptors in adenocarcinoma of the breast. Am J Pathol, 137, 863–70.

    Google Scholar 

  27. Pignatelli, M, Cardillo, MR, Hanby, A and Stamp, GWH, 1992, Integrins and their accessory adhesion molecules in mammary carcinomas: loss of polarization in poorly differentiated tumors. Hum Pathol, 23, 1159–66.

    Google Scholar 

  28. Zutter, MM, Krigman, HR and Santro, SA, 1993, Altered expression in adenocarcinoma of the breast. Analysis by in situ hybridization. Am J Pathol, 142, 1439–48.

    Google Scholar 

  29. Arihiro, K, Inai, K, Kurihara, K et al., 1993, Loss of VLA-2 collagen receptor in breast carcinoma, facilitating invasion and metastasis. Jpn J Cancer Res, 84, 726–33.

    Google Scholar 

  30. Mechtersheimer, G, Munk, M, Barth, T, Koretz, K and Möller, P, 1993, Expression of β1 integrins in non-neoplastic mammary epithelium, fibroadenoma and carcinoma of the breast. Virchows Archiv A, 422, 203–10.

    Google Scholar 

  31. Berdichevsky, F, Gilbert, C, Shearer, M and Taylor-Papadimitriou, J, 1992, Collagen-induced rapid morphogenesis of human mammary epithelial cells: The role of the α2β1 integrin. J Cell Sci, 102, 437–46.

    Google Scholar 

  32. Zutter, MM and Santro, SA, 1990, Widespread histologic distribution of the 012131 integrin cell-surface collagen receptor. Am J Pathol, 137, 113–20.

    Google Scholar 

  33. 33. Staatz, WD, Rajpara, SM, Wayner, EA, Carter, WG and Santoro, SA, 1989, The membrane glycoprotein Ia–IIa (VLA-2) complex mediates the Mg++-dependent adhesion of platelets to collagen. J Cell Biol, 108, 1917–24.

    Google Scholar 

  34. Elices, MJ and Hemler, ME, 1989, The human integrin VLA-2 is a collagen receptor on some cells and a collagen/laminin receptor on others. Proc Natl Acad Sci USA, 86, 9906–10.

    Google Scholar 

  35. Kirchhofer, D, Languino, LR, Ruoslahti, E and Pierschbacher, MD, 1990, α2β1 integrins from different cell types show different binding specificities. J Biol Chem, 265, 615–8.

    Google Scholar 

  36. Languino, LR, Gehlsen, KR, Wayner, E et al., 1989, Endothelial cells use α2β1 integrin as a laminin receptor. J Cell Biol, 109, 2455–62.

    Google Scholar 

  37. Kramer, RH and Marks, N, 1989, Identification of integrin collagen receptors on human melanoma cells. J Biol Chem, 264, 4684–8.

    Google Scholar 

  38. Lampugnani, MG, Resnati, M, Dejana, E and Marchisio, PC, 1991, The role of integrins in the maintenance of endothelial monolayer integrity. J Cell Biol, 112, 479–90.

    Google Scholar 

  39. Larjava, H, Peltonen, J, Akiyama, SK et al., 1990, Novel function for β1 integrins in keratinocyte cell-cell interactions. J Cell Biol, 110, 803–15.

    Google Scholar 

  40. Akiyama, SK, Yamada, SS, Chen, W-T and Yamada, KM, 1989, Analysis of fibronectin receptor function with monoclonal antibodies: Roles in cell adhesion, migration, matrix assembly, and cytoskeletal organization. J Cell Biol, 109, 863–75.

    Google Scholar 

  41. Pischel, KD, Hemler, ME, Huang, C, Bluestein, HG and Woods Jr., VL, 1987, Use of the monoclonal antibody 12F1 to characterize the differentiation antigen VLA-2. J Immunol, 138, 226–33.

    Google Scholar 

  42. Coller, BS, Beer, JH, Scudder, LE and Steinberg, MH, 1989, Collagen-platelet interactions: evidence for a direct interaction of collagen with platelet GPIa/IIa and an indirect interaction with platelet GPIIb/IIIa mediated by adhesive proteins. Blood, 74, 182–92.

    Google Scholar 

  43. Thompson, EW, Brünner, N, Torri, J et al., 1993, The invasive and metastatic properties of hormone-independent but hormone-responsive variants of MCF-7 human breast cancer cells. Clin Exp Metastasis, 11, 15–26.

    Google Scholar 

  44. Clark, R, Stampfer, MR, Milley, R et al., 1988, Transformation of human mammary epithelial cells by oncogenic retroviruses. Cancer Res, 48, 4689–94.

    Google Scholar 

  45. Soule, HD, Maloney, TM, Wolman, SR et al., 1990, Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res, 50, 6075–86.

    Google Scholar 

  46. Engel, LW and Young, NA, 1978, Human breast carcinoma cells in continuous culture: a review. Cancer Res, 38, 4327–39.

    Google Scholar 

  47. Thompson, EW, Paik, S, Brünner, N et al., 1992, Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol, 150, 534–44.

    Google Scholar 

  48. Sommers, CL, Thompson, EW, Torri, JA et al., 1991, Cell adhesion molecule uvomorulin expression in human breast cancer cell lines: relationship to morphology and invasive capacities. Cell Growth Diff, 2, 365–72.

    Google Scholar 

  49. Sommers, CL, Walker-Jones, D, Heckford, SE et al., 1989, Vimentin rather than keratin expression in some hormone-independent breast cancer cell lines and in oncogene-transformed mammary epithelial cells. Cancer Res, 49, 4258–63.

    Google Scholar 

  50. Boyer, B, Tucker, GC, Vallés, AM, Franke, WW and Thiery, JP, 1989, Rearrangements of desmosomal and cytoskeletal proteins during the transition from epithelial to fibroblastoid organization in cultured rat bladder carcinoma cells. J Cell Biol, 109, 1495–509.

    Google Scholar 

  51. Raymond, WA and Leong, AS-Y, 1989, Vimentin — a prognostic parameter in breast carcinoma? J Pathol, 158, 107–14.

    Google Scholar 

  52. Raymond, WA and Leong, AS-Y, 1989, Co-expression of cytokeratin and vimentin intermediate filament proteins in benign and neoplastic breast epithelium. J Pathol, 157, 299–306.

    Google Scholar 

  53. Rao, CN, Castronovo, V, Schmitt, MC et al., 1989, Evidence for a precursor of the high-affinity metastasis-associated murine laminin receptor. Biochem, 28, 7476–86.

    Google Scholar 

  54. Timpl, R, Rohde, H, Robey, PG et al., 1979, Laminin — a glycoprotem from basement membrane. J Biol Chem, 254, 9933–7.

    Google Scholar 

  55. Malinoff, HL, McCoy Jr., JP, Varani, J and Wicha, MS, 1984, Metastatic potential of murine fibrosarcoma cells is influenced by cell surface laminin. Int J Cancer, 33, 651–5.

    Google Scholar 

  56. Terranova, VP, Williams, JE, Liotta, LA and Martin, GR, 1984, Modulation of the metastatic activity of melanoma cells by laminin and fibronectin. Science, 226, 982–5.

    Google Scholar 

  57. Juliano, RL and Haskill, S, 1993, Signal transduction from the extracellular matrix. J Cell Biol, 120, 577–85.

    Google Scholar 

  58. Terpe, H-J, Tajrobehkar, K, Günthert, U and Altmannsberger, M, 1993, Expression of cell adhesion molecules alpha-2, alpha-5 and alpha-6 integrin, E-cadherin, N-CAM and CD-44 in renal cell carcinomas. An immunohistochemical study. Virchows Archiv A, 422, 219–24.

    Google Scholar 

  59. Favrot, MC, Combaret, V, Goillot, E et al., 1991, Expression of integrin receptors on 45 clinical neuroblastoma specimens. Int J Cancer, 49, 347–55.

    Google Scholar 

  60. Jones, J, Sugiyama, M, Watt, FM and Speight, PM, 1993, Integrin expression in normal, hyperplastic, dysplastic, and malignant oral epithelium. J Pathol, 169, 235–43.

    Google Scholar 

  61. Chan, BMC, Matsuura, N, Takada, Y, Zetter, B and Hemler, ME, 1991, In vitro and in vivo consequences of VLA-2 expression on rhabdomyosarcoma cells. Science, 251, 1600–2.

    Google Scholar 

  62. Nicolson, GL, 1987, Tumor cell instability, diversification, and progression to the metastatic phenotype: from oncogene to oncofetal expression. Cancer Res, 47, 1473–87.

    Google Scholar 

  63. Heino, J and Massagué, J, 1989, Transforming growth factor-β switches the pattern of integrins expressed in MG-63 human osteosarcoma cells and causes a selective loss of cell adhesion to laminin. J Biol Chem, 264, 21806–11.

    Google Scholar 

  64. Arrick, BA, Lopez, AR, Elfman, F et al., 1992, Altered metabolic and adhesive properties and increased tumorigenesis associated with increased expression of transforming growth factor β1. J Cell Biol, 118, 715–26.

    Google Scholar 

  65. Klein, K, Giancotti, FG, Presta, M et al., 1993, Basic fibroblast growth factor modulates integrin expression in microvascular endothelial cells. Mol Biol Cell, 4, 973–82.

    Google Scholar 

  66. Santala, P and Heino, J, 1991, Regulation of integrin-type cell adhesion receptors by cytokines. J Biol Chem, 266, 23505–9.

    Google Scholar 

  67. Defilippi, P, Silengo, L and Tarone, G, 1992, α6β1 integrin (laminin receptor) is down-regulated by tumor necrosis factor a and interleukin-1 β in human endothelial cells. J Biol Chem, 267, 18303–7.

    Google Scholar 

  68. Dedhar, S, 1989, Regulation of expression of the cell adhesion receptors, integrins, by recombinant human interleukin-1β in human osteosarcoma cells: inhibition of cell proliferation and stimulation of alkaline phosphatase activity. J Cell Physiol, 138, 291–9.

    Google Scholar 

  69. Conforti, G, Zanetti, A, Pasquali-Ronchetti, I et al., 1990, Modulation of vitronectin receptor binding by membrane lipid composition. J Biol Chem, 265, 4011–9.

    Google Scholar 

  70. Kirchhofer, D, Grzesiak, J and Pierschbacher, MD, 1991, Calcium as a potential physiological regulator of integrin-mediated cell adhesion. J Biol Chem, 266, 4471–7.

    Google Scholar 

  71. Lampe, BV, Stallmach, A and Riecken, EO, 1993, Altered glycosylation of integrin adhesion molecules in colorectal cancer cells and decreased adhesion to the extracellular matrix. Gut, 34, 829–36.

    Google Scholar 

  72. Tamura, RN, Cooper, HM, Collo, G and Quaranta, V, 1991, Cell type-specific integrin variants with alternative a chain cytoplasmic domain. Proc Natl Acad Sci USA, 88, 10183–7.

    Google Scholar 

  73. Du, X, Plow, EF, Frelinger III, AL et al., 1991, Ligands ‘activate’ integrin αIIbβ3 (platelet GPIIb–IIIa). Cell, 65, 409–16.

    Google Scholar 

  74. Chan, BMC and Hemler, ME, 1993, Multiple functional forms of the integrin VLA-2 can be derived from a single α2 cDNA clone: interconversion of forms induced by an anti-β1 antibody. J Cell Biol, 120, 537–43.

    Google Scholar 

  75. Witkowski, CM, Rabinovitz, I, Nagle, RB, Affinito, KS and Cress, AE, 1993, Characterization of integrin subunits, cellular adhesion and tumorigenicity of four human prostate cell lines. J Cancer Res Clin Oncol, 119, 637–44.

    Google Scholar 

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Authors and Affiliations

  1. Lombardi Cancer Research Center, Georgetown University, Washington, DC, USA

    Michio Maemura & Robert B. Dickson

  2. Laboratory of Developmental Biology, National Institute of Dental Research, Bethesda, MD, USA

    Steven K. Akiyama

  3. Division of Rheumatic Diseases, School of Medicine, University of California, San Diego, CA, USA

    Virgil L. Woods Jr.

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  1. Michio Maemura
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  2. Steven K. Akiyama
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  3. Virgil L. Woods Jr.
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  4. Robert B. Dickson
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Maemura, M., Akiyama, S.K., Woods, V.L. et al. Expression and ligand binding of α2β1 integrin on breast carcinoma cells. Clin Exp Metast 13, 223–235 (1995). https://doi.org/10.1007/BF00133478

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