, Volume 4, Issue 4, pp 269–275 | Cite as

Fibrin fragment E stimulates the proliferation, migration and differentiation of human microvascular endothelial cells in vitro

  • C.A. Bootle-Wilbraham
  • S. Tazzyman
  • W.D. Thompson
  • C.M. Stirk
  • C.E. Lewis


Various factors involved in haemostasis also regulate the development of new blood vessels by a process called angiogenesis. Enzymatic cleavage of fibrin yields a variety of fibrin degradation products, particularly in areas of intense angiogenesis such as in healing wounds and active atherosclerotic plaques. One of these, fibrin fragment E (FnE), is a potent angiogenic factor in the chick chorioallantoic membrane assay of angiogenesis. Here, we extend these studies to show that FnE stimulates the proliferation, migration and differentiation of human dermal microvascular endothelial cells (HuDMECs) in vitro, both in the absence and presence of such additional endothelial growth factors as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). We also show that these stimulatory effects occur at concentrations of the protein known to be present in angiogenic tissues in vivo. FnE enhanced the angiogenic effects of VEGF or bFGF, indicating a possible synergy between the signalling pathways used by these three angiogenic factors.

angiogenesis endothelial cell fibrin fibrinolysis fragment E Matrigel 


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  1. 1.
    Browder T, Folkman J, Pirie-Shepherd S. The hemostatic system as a regulator of angiogenesis. J Biol Chem 2000; 275: 1521–4.PubMedCrossRefGoogle Scholar
  2. 2.
    Hawiger J. Formation and regulation of platelet and fibrin haemostatic plug. Hum Pathol 2000; 18: 111–22.Google Scholar
  3. 3.
    Doolittle RF. Fibrinogen and fibrin. Sci Am 1981; 245: 92–101.CrossRefGoogle Scholar
  4. 4.
    Chafer BM, Maier K, Eickhoff U et al. Plasminogen activation in healing human wounds. Am J Pathol 1994; 144: 1269–80.Google Scholar
  5. 5.
    Sahni A, Odrljin T, Francis CW. Binding of basic fibroblast growth factor to fibrinogen and fibrin. J Biol Chem 1998; 273: 7554–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Pepper MS, Sappino AP, Stocklin R et al. Upregulation of urokinase receptor expression on migrating endothelial cells. J Biol Chem 1993; 122: 673–84.Google Scholar
  7. 7.
    Brooks PC, Clark RA, Cheresh DA. Requirement of vascular integrin alpha v beta 3 for angiogenesis. Science 1994; 264: 569–71.PubMedGoogle Scholar
  8. 8.
    Bayless KJ, Salazar R, Davis GE. RGD-Dependent vacuolation and lumen formation observed during endothelial cell morphogenesis in three-dimensional fibrin matrices involves the avb3 and a5b1 integrins. Am J Pathol 2000; 156: 1673–83.PubMedGoogle Scholar
  9. 9.
    Smith EB, Keen GA, Grant A, Stirk C. Fate of fibrinogen in human arterial intima. Arteriosclerosis 1990; 10: 263–5.PubMedGoogle Scholar
  10. 10.
    Thompson WD, Smith EB, Stirk CM, et al. Angiogenic activity of fibrin degradation products is located in fibrin fragment. EJ Pathol 1992; 168: 47–53.CrossRefGoogle Scholar
  11. 11.
    Naito M, Stirk CM, Smith EB, Thompson WD. Smooth Muscle cell outgrowth stimulated by fibrin degradation products: The potential role of fibrin fragment Ein restenosis and atherogenesis. Thromb Res 2000; 98: 165–74.PubMedCrossRefGoogle Scholar
  12. 12.
    Kumar R, Yoneda J, Bucana CD, Fidler IJ. Regulation of distinct steps of angiogenesis by different angiogenic molecules. Int J Oncol 1998; 12: 749–57.PubMedGoogle Scholar
  13. 13.
    Mullin JL, Gorkun OV, Binnie CG, Lord ST. Recombinant fibrinogen studies reveal that thrombin specificity dictates order of fibrinopeptide release. J Biol Chem 2000; 275: 25246–69.CrossRefGoogle Scholar
  14. 14.
    Liu CY, Soberl JH, Weitz JI et al. Immunologic identification of the cleavage products from the A alpha-and B beta-chains in the early stages of plasmin digestion of fibrinogen. Thromb Haemost 1986; 56: 100–6.PubMedGoogle Scholar
  15. 15.
    Liu J, Kolath J, Anderson J, et al. Positive interaction between 5–FU and FdUMP[10] in the inhibition of human colorectal tumour cell proliferation. Antisense Nucleic Acid Drug Dev 1999; 9: 481–6.PubMedGoogle Scholar
  16. 16.
    Malinda KM, Ponce L, Kleinman HK et al. Gp38k, a protein synthesized by vascular smooth muscle cells, stimulates directional migration of human umbilical vein endothelial cells. Exp Cell Res 1999; 250: 168–73.PubMedCrossRefGoogle Scholar
  17. 17.
    Shen J, Ham RG, Karmiol S. Expression of adhesion molecules in cultured human pulmonary microvascular endothelial cells. Microvasc Res 1995; 50: 360–72.PubMedCrossRefGoogle Scholar
  18. 18.
    Gando S, Tedo I, Kubota M. Posttrauma coagulation and fibrinolysis. Crit Care Med 1992; 20: 594–600.PubMedGoogle Scholar
  19. 19.
    Tanaka K, Abe M, Sato Y. Roles of extracellular signal regulated kinase \(\frac{1}{2}\) and p38 mitogen-activated protein kinase in the signal transduction of basic fibroblast growth factor in endothelial cells during angiogenesis. Jpn J Cancer Res 1999; 90: 647–54.PubMedGoogle Scholar
  20. 20.
    Rousseau S, Houle F, Landry J, Huot J. p38 MAP kinase activation by vascular endothelial growth factor mediates actin reorganisation and cell migration in human endothelial cells. Oncogene 1997; 15: 2169–77.PubMedCrossRefGoogle Scholar
  21. 21.
    Sahni A, Sporn LA, Francis CW. Potentiation of endothelial cell proliferation by fibrin(ogen) bound fibroblast growth factor-2. J Biol Chem 1999; 274: 14936–41.PubMedCrossRefGoogle Scholar
  22. 22.
    Sahni A, Baker CA, Sporn LA, Fancis CW. Fibrinogen and Fibrin protect fibrobalst growth factor-2 from proteolytic degradation. Thromb Haemost 2000; 83: 736–41.PubMedGoogle Scholar
  23. 23.
    Sahni A, Francis CW. Vascular endothelial growth factor binds to fibrinogen and fibrin and stimulates endothelial cell proliferation. Blood 2000; 96: 3772–8.PubMedGoogle Scholar
  24. 24.
    Belgore FM, Blann AD, Lip GY. Measurement of free and complexed soluble vascular endothelial growth factor receptor, Flt-1, in fluid samples: Development and application of two new immunoassays. Clin Sci (Colch) 2001 May; 100(5): 567–75.CrossRefGoogle Scholar
  25. 25.
    Okunieff P, Barrett AJ, Phang SE et al. Circulating basic fibroblast growth factor declines during Cy/TBI bone marrow transplantation. Bone Marrow Transplant 1999; 23(11): 1117–21.PubMedCrossRefGoogle Scholar
  26. 26.
    Thiagarajan P, Rippon AJ, Farrell DH. Alternative adhesion sites in human fibrinogen for vascular endothelial cells. Biochemistry 1996; 35: 4169–75.PubMedCrossRefGoogle Scholar
  27. 27.
    Francis CW, Bunce LA, Sporn LA. Endothelial cell responses to fibrin mediated by FPB cleavage and the amino terminus of the beta chain. Blood Cells 1993; 19: 291–306.PubMedGoogle Scholar
  28. 28.
    Elicieri BP, Cheresh DA. Role of alpha v integrins during angiogenesis. Cancer J Sci Am 2000; 3: S245–9.Google Scholar
  29. 29.
    Bootle-Wilbraham CA, Tazzyman S, Marshall JM, Lewis CE. Fibrinogen E-fragment inhibits the migration and tubule formation of human dermal microvascular endothelial cells in vitro. Cancer Research 2000; 60: 4719–24.PubMedGoogle Scholar
  30. 30.
    Dejana E, Languino LR, Polentarutti N et al. Interaction between fibrinogen and cultured endothelial cells. Induction of migration and specific binding. J Clin Invest 1985; 75: 11–8.PubMedCrossRefGoogle Scholar
  31. 31.
    Burgos H, Herd A, Bennett JP. Placental angiogenic and growth factors in the treatment of chronic varicose ulcers: Preliminary communication. J R Soc Med 1989; 82: 598–9.PubMedGoogle Scholar
  32. 32.
    Pecher P, Schumacher BA. Angiogenesis in ischemic human myocardium: Clinical results after 3 years. Ann Thorac Surg 2000; 69: 1414–9.PubMedCrossRefGoogle Scholar
  33. 33.
    Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nature Med 2000; 6: 389–95.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • C.A. Bootle-Wilbraham
    • 1
  • S. Tazzyman
    • 1
  • W.D. Thompson
    • 2
  • C.M. Stirk
    • 3
  • C.E. Lewis
    • 4
  1. 1.Tumour Targeting Group, Section of Pathology, Division of Genomic MedicineUniversity of Sheffield Medical SchoolSheffieldUK
  2. 2.Department of PathologyUniversity of Aberdeen Medical School, Aberdeen Royal InfirmaryAberdeenUK
  3. 3.Molecular and Cell BiologyUniversity of Aberdeen Medical School, Aberdeen Royal InfirmaryAberdeenUK
  4. 4.Tumour Targeting Group, Section of Pathology, Division of Genomic MedicineUniversity of Sheffield Medical SchoolSheffieldUK. Tel/Fax:

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