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Thromb Haemost 2001; 86(02): 616-622
DOI: 10.1055/s-0037-1616095
Review Article
Schattauer GmbH

High Affinity Binding of Heparin by Necrotic Tumour Cells Neutralises Anticoagulant Activity

Implications for Cancer Related Thromboembolism and Heparin Therapy
Sumihito Morita   *
1   Thrombosis Research Institute, London and Imperial College School of Medicine, Hammersmith Hospital, London, UK
,
Milena A. Gebska
1   Thrombosis Research Institute, London and Imperial College School of Medicine, Hammersmith Hospital, London, UK
,
Ajay K. Kakkar
1   Thrombosis Research Institute, London and Imperial College School of Medicine, Hammersmith Hospital, London, UK
,
Michael F. Scully
1   Thrombosis Research Institute, London and Imperial College School of Medicine, Hammersmith Hospital, London, UK
› Author Affiliations
Further Information

Publication History

Received 13 November 2000

Accepted after resubmission 20 January 2001

Publication Date:
12 December 2017 (online)

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Summary

We have observed a striking neutralisation of the anticoagulant activity of unfractionated heparin in the presence of a pancreatic carcinoma cell line (MIA PaCa-2) due to binding of around 9 g of heparin per 107 cells (apparent Kd, 30 nM). The loss of anticoagulant activity was less marked in the presence of low molecular weight forms of heparin. Binding to the cell blocked acceleration of the thrombin:anti-thrombin interaction by heparin. Neutralisation of heparin activity was also shown to occur in the presence of a number of other tumour cell lines. FACS analysis demonstrated that live cells did not bind heparin and high affinity binding only occurred to dead MIA PaCa-2 cells. Heparin binding proteins accumulating in cell medium were identified as histone and ribosomal proteins that will become exposed during necrosis. The release of these proteins from cells within the necrotic core of a tumour or from cells killed during chemotherapy may abrogate the heparan sulphate/antithrombin system and possibly contribute to the idiopathic thromboembolism often associated with cancer (Trousseau’s syndrome). The findings also suggest a reason for the reported advantage of LMWH over UFH in treating venous thromboembolism in cancer patients and in improving patient survival.

Keywords

Heparin binding - cancer - thromboembolism

* Present address: Dr. S. Morita, Hamamatsu University School of Medicine, Shizuoka-ken, Japan


 

  • References

  • 1 Camerer E, Kolsto AB, Prydz H. Cell biology of tissue factor, the principal initiator of blood coagulation. Thromb Res 1996; 81: 1-41.
    CrossrefPubMedGoogle Scholar
  • 2 Dvorak HF. Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing, N Engl J Med. 1986; 315: 1650-9.
    PubMedGoogle Scholar
  • 3 Nierodzik ML, Plotkin A, Kajumo F, Karpatkin S. Thrombin stimulates tumor-platelet adhesion in vitro and metastasis in vivo. J Clin Invest 1991; 87: 229-36.
    CrossrefPubMedGoogle Scholar
  • 4 Nierodzik ML, Kajumo F, Karpatkin S. Effect of thrombin treatment of tumor cells on adhesion of tumor cells to platelets in vitro and tumor metastasis in vivo. Cancer Res 1992; 52: 3267-72.
    PubMedGoogle Scholar
  • 5 Nierodzik ML, Chen K, Takeshita K, Li JJ, Huang YQ, Feng XS, D’Andrea MR, Andrade-Gordon P, Karpatkin S. Protease-activated receptor 1 (PAR-1) is required and rate-limiting for thrombin-enhanced experimental pulmonary metastasis. Blood 1998; 92: 3694-700.
    PubMedGoogle Scholar
  • 6 Zain J, Huang YQ, Feng X, Nierodzik ML, Li JJ, Karpatkin S. Concentration-dependent dual effect of thrombin on impaired growth/apoptosis or mito-genesis in tumor cells. Blood 2000; 95: 3133-8.
    CrossrefPubMedGoogle Scholar
  • 7 Biggerstaff JP, Seth N, Amirkhosravi A, Amaya M, Fogarty S, Meyer TV, Siddiqui F, Francis JL. Soluble fibrin augments platelet/tumor cell adherence in vitro and in vivo, and enhances experimental metastasis. Clin Exp Metastasis 1999; 17: 723-30.
    CrossrefPubMedGoogle Scholar
  • 8 Zacharski LR, Ornstein DL. Heparin and cancer. Thromb Haemost 1998; 80: 10-23.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Siragusa S, Cosmi B, Piovella F, Hirsh J, Ginsberg JS. Low-molecular-weight heparins and unfractionated heparin in the treatment of patients with acute venous thromboembolism: results of a meta-analysis. Am J Med 1996; 100: 269-77.
    CrossrefPubMedGoogle Scholar
  • 10 Lensing AW, Prins MH, Davidson BL, Hirsh J. Treatment of deep venous thrombosis with low-molecular-weight heparins. A meta-analysis. Arch Intern Med 1995; 155: 601-7.
    CrossrefPubMedGoogle Scholar
  • 11 Engelberg H. Actions of heparin that may affect the malignant process. Cancer 1999; 85: 257-72.
    CrossrefPubMedGoogle Scholar
  • 12 Hileman RE, Fromm JR, Weiler JM, Linhardt RJ. Glycosaminoglycan-protein interactions: definition of consensus sites in glycosaminoglycan binding proteins. Bioessays 1998; 20: 156-67.
    CrossrefPubMedGoogle Scholar
  • 13 Young E, Prins M, Levine MN, Hirsh J. Heparin binding to plasma proteins, an important mechanism for heparin resistance. Thromb Haemost 1992; 67: 639-43.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 de Swart CA, Nijmeyer B, Roelofs JM, Sixma JJ. Kinetics of intravenously administered heparin in normal humans. Blood 1982; 60: 1251-8.
    PubMedGoogle Scholar
  • 15 Boneu B, Buchanan MR, Caranobe C, Gabaig AM, Dupouy D, Sie P, Hirsh J. The disappearance of a low molecular weight heparin fraction (CY 216) differs from standard heparin in rabbits. Thromb Res 1987; 46: 845-53.
    CrossrefPubMedGoogle Scholar
  • 16 Beguin S, Lindhout T, Hemker HC. The effect of trace amounts of tissue factor on thrombin generation in platelet rich plasma, its inhibition by heparin. Thromb Haemost 1989; 61: 25-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Doucet JP, Trifaro JM. A discontinous and highly porous sodium dodecyl sulfate-polyacrylamide slab gel system of high resolution. Anal Biochem 1988; 168: 265-71.
    CrossrefPubMedGoogle Scholar
  • 18 Bilozur ME, Biswas C. Identification and characterization of heparan sul-fate-binding proteins from human lung carcinoma cells. J Biol Chem 1990; 265: 19697-703.
    PubMedGoogle Scholar
  • 19 Lane DA, Denton J, Flynn AM, Thunberg L, Lindahl U. Anticoagulant activities of heparin oligosaccharides and their neutralization by platelet factor 4. Biochem J 1984; 218: 725-32.
    CrossrefPubMedGoogle Scholar
  • 20 Lane DA, Pejler G, Flynn AM, Thompson EA, Lindahl U. Neutralization of heparin-related saccharides by histidine-rich glycoprotein and platelet factor 4. J Biol Chem 1986; 261: 3980-6.
    PubMedGoogle Scholar
  • 21 Castellot Jr JJ, Wong K, Herman B, Hoover RL, Albertini DF, Wright TC, Caleb BL, Karnovsky MJ. Binding and internalization of heparin by vascular smooth muscle cells. J Cell Physiol 1985; 124: 13-20.
    CrossrefPubMedGoogle Scholar
  • 22 Kjellen L, Oldberg A, Hook M. Cell-surface heparan sulfate. Mechanisms of proteoglycan-cell association, J Biol Chem 1980; 255: 10407-13.
    PubMedGoogle Scholar
  • 23 Glimelius B, Busch C, Hook M. Binding of heparin on the surface of cultured human endothelial cells. Thromb Res 1978; 12: 773-82.
    CrossrefPubMedGoogle Scholar
  • 24 Biswas C. Heparin and heparan sulfate binding sites on B-16 melanoma cells. J Cell Physiol 1988; 136: 147-53.
    CrossrefPubMedGoogle Scholar
  • 25 Leung L, Saigo K, Grant D. Heparin binds to human monocytes and modulates their procoagulant activities and secretory phenotypes. Effects of histidine-rich glycoprotein. Blood 1989; 73: 177-84.
    PubMedGoogle Scholar
  • 26 Raboudi N, Julian J, Rohde LH, Carson DD. Identification of cell-surface heparin/heparan sulfate-binding proteins of a human uterine epithelial cell line (RL95). J Biol Chem 1992; 267: 11930-9.
    PubMedGoogle Scholar
  • 27 Hoke DE, Regisford EG, Julian J, Amin A, Begue-Kirn C, Carson DD. Murine HIP/L29 is a heparin-binding protein with a restricted pattern of expression in adult tissues. J Biol Chem 1998; 273: 25148-57.
    CrossrefPubMedGoogle Scholar
  • 28 Watson K, Gooderham NJ, Davies DS, Edwards RJ. Nucleosomes bind to cell surface proteoglycans. J Biol Chem 1999; 274: 21707-13.
    CrossrefPubMedGoogle Scholar
  • 29 Lane DA. Heparin binding and neutralising proteins. In: Heparin. Chemical and Biological Properties, Clinical Applications. Lane DA, Lindahl U. (eds.) London: Edward Arnold; 1989: 363-92.
    Google Scholar
  • 30 Callander N, Rapaport SI. Trousseau’s syndrome. West J Med 1993; 158: 364-71.
    PubMedGoogle Scholar
  • 31 Soini Y, Paakko P, Lehto VP. Histopathological evaluation of apoptosis in cancer. Am J Pathol 1998; 153: 1041-53.
    CrossrefPubMedGoogle Scholar
  • 32 Meggiato T, Calabrese F, Valente M, Favaretto E, Baliello E, Del Favero G. Spontaneous apoptosis and proliferation in human pancreatic cancer. Pancreas. 2000; 20: 117-22.
    CrossrefPubMedGoogle Scholar
  • 33 Majno G, Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 1995; 146: 3-15.
    PubMedGoogle Scholar
  • 34 Leist M, Nicotera P. The shape of cell death. Biochem Biophys Res Commun 1997; 236: 1-9.
    CrossrefPubMedGoogle Scholar
  • 35 Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol 1980; 68: 251-306.
    PubMedGoogle Scholar
  • 36 Brown JM, Giaccia AJ. The unique physiology of solid tumors: opportunities (and problems) for cancer therapy. Cancer Res 1998; 8: 1408-16.
    PubMedGoogle Scholar
  • 37 Greeno EW, Bach RR, Moldow CF. Apoptosis is associated with increased cell surface tissue factor procoagulant activity. Lab Invest 1996; 75: 281-9.
    PubMedGoogle Scholar
  • 38 Rosenberg RD, Rosenberg JS. Natural anticoagulant mechanisms. J Clin Invest 1984; 74: 1-6.
    CrossrefPubMedGoogle Scholar
  • 39 Stein PE, Carrell RW. What do dysfunctional serpins tell us about molecular mobility and disease?. Nat Struct Biol 1995; 2: 96-113.
    CrossrefPubMedGoogle Scholar
  • 40 Falanga A. Mechanisms of hypercoagulation in malignancy and during chemotherapy. Haemost 1998; 28 (Suppl. 03) 50-60.
    CrossrefPubMedGoogle Scholar