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Thromb Haemost 2003; 90(02): 351-360
DOI: 10.1160/TH03-02-0088
Cellular Proteolysis and Oncology
Schattauer GmbH

Inhibition of thrombosis in melanoma allografts in mice by endogenous mast cell heparin

Michael Samoszuk
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
,
Mark Corwin
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
,
Hon Yu
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
,
Jun Wang
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
,
Orhan Nalcioglu
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
,
Min-Ying Su
1   Departments of Pathology, Physics, Radiologic Sciences and Center for Functional Onco-Imaging, University of California, Irvine, USA
› Author AffiliationsFinancial support: This work was supported by grants from California Cancer Research program
Further Information

Publication History

Received 07 February 2003

Accepted after resubmission 18 May 2003

Publication Date:
06 December 2017 (online)

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Summary

An unexplained paradox of malignant melanoma is the apparent failure of the blood within the tumor to clot despite the presence of multiple factors that should promote blood clotting. Here we present histochemical evidence that human and murine melanomas are extensively infiltrated by abundant mast cells. Because mast cells contain the natural anticoagulant heparin, the present studies were aimed at defining the role of mast cell heparin in preventing the blood from clotting within B16 melanoma grafts in C57BL/6 J mice. Mice bearing B16 melanoma grafts were treated with non-specific or specific inhibitors of mast cell heparin (protamine or heparinase, respectively). After the drug treatment there was histologic and functional evidence of selective thrombosis of the blood vessels within the protamine and heparinase treated melanoma grafts. A similar, high degree of thrombosis was also observed in B16 tumors grown in transgenic NDST-2 knockout mice bearing a targeted disruption in the gene coding for mast cell heparin synthesis. The tumors grown in the protamine-treated animals were significantly smaller than the tumors from control (untreated mice). By contrast, the tumors treated with heparinase or grown in the NDST-2 knockout mice were significantly larger than the tumors from control (untreated) mice. We conclude that the intrinsic procoagulant properties of malignant melanoma are neutralized in vivo by the anticoagulant properties of endogenous heparin produced by mast cells that naturally infiltrate the tumor. Our results also suggest that thrombosis and hemostasis within melanoma may play a complex role in modulating the growth of the tumor.

Keywords

Mast cell - heparin - cancer - coagulation
 

  • References

  • 1 Baish JW, Netti PA, Jain RK. Transmural coupling of fluid in microcirculatory network and interstitium in tumors. Microvasc Res 1997; 53: 128-41.
    CrossrefPubMedGoogle Scholar
  • 2 Hashizume H, Baluk P, Morikawa S. et al. Openings between defective endothelial cells explain tumor vessel leakiness. Am. J. Pathol 2000; 156: 1363-80.
    PubMedGoogle Scholar
  • 3 Stohrer M, Boucher Y, Stangassinger M, Jain RK. Oncotic pressure in solid tumors is elevated. Cancer Res 2000; 60: 4251-5.
    PubMedGoogle Scholar
  • 4 Chang YS, di Tomaso E, McDonald DM. et al. Mosaic blood vessels in tumors: frequency of cancer cells in contact with flowing blood. Proc Nat Acad Sci 2000; 97: 14608-13.
    CrossrefPubMedGoogle Scholar
  • 5 Siddiqui FA, Amirkhosravi A, Amaya M. et al. Purification and properties of human melanoma cell tissue factor. Clin Appl Thromb Hemos 2001; 7: 289-95.
    CrossrefPubMedGoogle Scholar
  • 6 Bromberg ME, Bailly MA, Konigsberg WH. Role of protease-activated receptor 1 in tumor metastasis promoted by tissue factor. Thromb. Haemost 2001; 86: 1210-4.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Amirkhosravi A, Biggerstaff JP, Warnes G. et al. Determination of tumor cell procoagulant activity by sonoclot analysis in whole blood. Thromb Res 1996; 84: 323-32.
    CrossrefPubMedGoogle Scholar
  • 8 Wotukiewicz MZ, Zacharski LR, Memoli VA. et al. Malignant melanoma. Interaction with coagulation and fibrinolysis pathways in situ. Am J Clin Pathol 1990; 93: 516-21.
    CrossrefPubMedGoogle Scholar
  • 9 Bottasso B, Mari D, Coppola R. et al. Hypercoagulability and hyperfibrinolysis in patients with melanoma. Thromb Res 1996; 81: 345-52.
    CrossrefPubMedGoogle Scholar
  • 10 Dvorak AM, Mihm MC, Osage Jr. JE, Dvorak HF. Melanoma. An ultrastructural study of the host inflammatory and vascular responses. J Invest Dermatol 1980; 75: 388-93.
    CrossrefPubMedGoogle Scholar
  • 11 Demitsu T, Inoue T, Kakurai M. et al. Activation of mast cells within a tumor of angiosarcoma: ultrastructural study of five cases. J Dermatol 2002; 29: 280-9.
    CrossrefPubMedGoogle Scholar
  • 12 Reed JA, McNutt NS, Bogndany JK, Albino AP. Expression of the mast cell growth factor interleukin-3 in melanocytic lesions correlates with an increased number of mast cells in the perilesional stroma: implications for melanoma progression. J Cutan Pathol 1996; 23: 495-505.
    CrossrefPubMedGoogle Scholar
  • 13 Duncan LM, Richards Jr. LA, Mihm MC. Increased mast cell density in invasive melanoma. J Cutan Pathol 1998; 25: 11-15.
    CrossrefPubMedGoogle Scholar
  • 14 Welker P, Schadendorf D, Artuc M, Grabbe J, Henz BM. Expression of SCF splice variants in human melanocytes and melanoma cell lines: potential prognostic implications. Br J Cancer 2000; 82: 1453-8.
    CrossrefPubMedGoogle Scholar
  • 15 Arber DA, Tamayo R, Weiss LM. Paraffin section detection of the c-kit gene product (CD117) in human tissues: value in the diagnosis of mast cell disorders. Hum Pathol 1998; 29: 498-504.
    CrossrefPubMedGoogle Scholar
  • 16 Starkey JR, Crowle PK, Taubenberger S. Mast-cell-defficient W/Wv mice exhibit a decreased rate of tumor angiogenesis. Int J Cancer 1988; 42: 48-52.
    CrossrefPubMedGoogle Scholar
  • 17 Toth-Jakatics R, Jimi S, Takebayashi S, Kawamoto N. Cutaneous malignant melanoma: correlation between neovascularization and peritumor accumulation of mast cells overexpressing vascular endothelial growth factor. Hum Pathol 2000; 31: 955-60.
    CrossrefPubMedGoogle Scholar
  • 18 Valent P, Baghestanian M, Bankl HC. et al. New aspects in thrombosis research : possible role of mast cells as profibrinolytic and anti-thrombotic cells. Thromb Haemost 2002; 87: 786-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Kauhanen P, Kovanen PT, Reunala T, Lassila R. Effects of skin mast cells on bleeding time and coagulation activation at the site of platelet plug formation. Thromb Haemost 1998; 79: 843-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Tchougounova E, Pejler G. Regulation of extravascular coagulation and fibrinolysis by heparin-dependent mast cell chymase. FASEB J 2001; 14: 2763-5.
    PubMedGoogle Scholar
  • 21 Sillaber C, Baghestanian M, Bevec D. et al. The mast cell as site of tissue-type plasminogen activator expression and fibrinolysis. J Immunol 1999; 162: 1032-41.
    CrossrefPubMedGoogle Scholar
  • 22 Mallett SV, Cox DJA. Thromboelastography. Brit J Anaes 1992; 69: 307-13.
    CrossrefPubMedGoogle Scholar
  • 23 Kang YG, Martin DJ, Marquez J. et al. Intraoperative changes in blood coagulation and thromboelastographic monitoring in liver transplantation. Anesth Analg 1985; 64: 888-96.
    PubMedGoogle Scholar
  • 24 Susic D, Mandal AK, Kentera D. Heparin lowers the blood pressure in hypertensive rats. Hypertension 1982; 4: 681-5.
    CrossrefPubMedGoogle Scholar
  • 25 Mandal AK, Lyden TW, Saklayen MG. Heparin lowers blood pressure: biological and clinical perspectives. Kidney Int 1995; 47: 1017-22.
    CrossrefPubMedGoogle Scholar
  • 26 Stucker M, Horstmann I, Nuchel C. et al. Blood flow compared in benign melanocytic naevi, malignant melanomas and basal cell carcinomas. Clin Exp Dermatol 1999; 24: 107-11.
    CrossrefPubMedGoogle Scholar
  • 27 Su MY, Wang Z, Roth GM. et al. Pharmacokinetic changes induced by vaso-modulators in kidneys, livers, muscles, and implanted tumors in rats as measured by dynamic Gd-DTPA-enhanced MRI. Magn Reson Med 1996; 36: 868-77.
    CrossrefPubMedGoogle Scholar
  • 28 Su MY, Muhler A, Lao X, Nalcioglu O. Tumor characterization with dynamic contrast-enhanced MRI using contrast agents of various molecular weights. Magn Reson Med 1998; 39: 259-69.
    CrossrefPubMedGoogle Scholar
  • 29 Lohse DL, Linhardt RJ. Purification and char-acterization of heparin lyases from Flavo-bacterium heparinum. J Bio Chem 1992; 267: 24347-55.
    CrossrefPubMedGoogle Scholar
  • 30 Su MY, Samoszuk MK, Wang J, Nalcioglu O. Assessment of protamine-induced thrombosis of tumor vessels for cancer therapy using dynamic contrast-enhanced MRI. NMR Biomed 2002; 15: 106-13.
    CrossrefPubMedGoogle Scholar
  • 31 Forsberg E, Pejler G, Ringvall M. et al. Abnormal mast cells in mice deficient in a heparin-synthesizing enzyme. Nature 1999; 400: 773-8.
    CrossrefPubMedGoogle Scholar
  • 32 Samoszuk M, Corwin MA. Acceleration of tumor growth and peri-tumoral blood clotting by imatinib mesylate (Gleevec).Int J Cancer. (in press 2003)
    PubMedGoogle Scholar
  • 33 Mannucci PM. Hemostatic drugs. N Engl J Med 1998; 339: 245-51.
    CrossrefPubMedGoogle Scholar
  • 34 Yano H, Kinuta M, Tateishi H. et al. Mast cell infiltration around gastric cancer cells correlates with tumor angiogenesis and metastasis. Gastric Cancer 1999; 1: 26-32.
    CrossrefPubMedGoogle Scholar
  • 35 Elpek GO, Gelen T, Aksoy NH. et al. The prognostic relevance of angiogenesis and mast cells in squamous cell carcinoma of the oesophagus. J Clin Pathol 2001; 54: 940-4.
    CrossrefPubMedGoogle Scholar
  • 36 Dethlefsen SM, Matsuura N, Zetter BR. Mast cell accumulation at sites of murine tumor implantation: implications for angiogenesis and tumor metastasis. Invasion Metastasis 1994-1995 14: 395-408.
    PubMedGoogle Scholar
  • 37 Cooussens LM, Raymond WW, Bergers G. et al. Inflammatory mast cells up-regulate angiogenesis during squamous epithelial carcinogenesis. Genes Dev 1999; 13: 1382-97.
    CrossrefPubMedGoogle Scholar
  • 38 Imada A, Shijubo N, Kojima H, Abe S. Mast cells correlate with angiogenesis and poor outcome in stage I lung adenocarcinoma. Eur Respir J 2000; 15: 1087-93.
    CrossrefPubMedGoogle Scholar
  • 39 Takanami I, Takeuchi K, Naruke M. Mast cell density is associated with angiogenesis and poor prognosis in pulmonary adenocarcinoma. Cancer 2000; 88: 2686-92.
    CrossrefPubMedGoogle Scholar
  • 40 Thomas VA, Wheeless CJ, Stack MS, Johnson DA. Human mast cell tryptase fibrinogenoly-sis: kinetics, anticoagulation mechanism, and cell adhesion disruption. Biochemistry 1998; 37: 2291-8.
    CrossrefPubMedGoogle Scholar
  • 41 Hallgren J, Estrada S, Karlson U, Alving K, Pejler G. Heparin antagonists are potent inhibitors of mast cell tryptase. Biochemistry 2001; 40: 7342-9.
    CrossrefPubMedGoogle Scholar
  • 42 Pejler G. Lactoferrin regulates the activity of heparin proteoglycan-bound mast cell chymase: characterization of the binding of heparin to lactoferrin. Biochem J 1996; 320: 897-903.
    CrossrefPubMedGoogle Scholar
  • 43 Samoszuk MK, Min-Ying S, Ahmad N, Nalcioglu O. Selective thrombosis of tumor blood vessels in mammary adenocarcinoma implants in rats. Am J Pathol 2001; 159: 245-51.
    CrossrefPubMedGoogle Scholar
  • 44 Ornstein DL, Zacharski LR. Treatment of cancer with anticoagulants: rationale in the treatment of melanoma. Int J Hematol 2001; 73: 157-61.
    CrossrefPubMedGoogle Scholar
  • 45 Desai UR, Wang H, Linhardt RJ. Substrate specificity of the heparin lyases from Flavobacterium heparinum. Arch Biochem Biophys 1993; 306: 461-8.
    CrossrefPubMedGoogle Scholar