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Thromb Haemost 1998; 80(04): 598-602
DOI: 10.1055/s-0037-1615428
Rapid Communication
Schattauer GmbH

Pentoxifylline Inhibits Hypoxia-induced Upregulation of Tumor Cell Tissue Factor and Vascular Endothelial Growth Factor

Ali Amirkhosravi
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Todd Meyer
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Gary Warnes
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Mildred Amaya
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Zaiba Malik
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
John P. Biggerstaff
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Farooq A. Siddiqui
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
Paul Sherman
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
,
John L. Francis
1   Cell Biology, Hemostasis and Thrombosis Research Unit, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando, Florida, USA
› Author Affiliations
Further Information

Publication History

Received 09 January 1998

Accepted after resubmission 30 June 1998

Publication Date:
08 December 2017 (online)

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Summary

Tissue factor (TF), the membrane glycoprotein that initiates blood coagulation, is constitutively expressed by many tumor cells and is implicated in peri-tumor fibrin deposition and hypercoagulability in cancer. Upregulation of tumor TF correlates with enhanced metastatic potential. Furthermore, TF has been colocalized with VEGF in breast cancer, specially at sites of early angiogenesis. There are no data on the effect of hypoxia on tumor cell TF expression. Since hypoxia is known to stimulate VEGF production, we studied whether this also induces tumor cell TF expression. Confluent monolayers of A375 melanoma, MCF-7 breast carcinoma and A549 lung carcinoma were cultured in either 95% air, 5% CO2 (normoxic) or 95% N2, 5% CO2 (hypoxic; 25-30 mmHg) for 24 h. Procoagulant activity (PCA) was measured by amidolytic and clotting assays, surface TF antigen by flow cytometry, early apoptosis by annexin V binding and VEGF levels in culture supernatants by ELISA. Hypoxia significantly increased tumor cell PCA in all three cell lines tested and TF antigen on A375 cells was increased four-fold (P <0.05). Pentoxifylline (PTX), a methylxanthine derivative, significantly inhibited the hypoxia-induced increase in PCA as well as VEGF release in all three cell lines tested. In A375 cells, PTX significantly inhibited TF antigen expression by both normoxic and hypoxic cells. Hypoxia induced a slight (5%) but not significant, increase in early apoptosis. Intravenous injection of hypoxic A375 cells into nude rats produced more pronounced thrombocytopenia (n = 5, P <0.01) and more lung metastases (n = 3, P <0.05) compared to normoxic cells. We conclude that hypoxia increases TF expression by malignant cells which enhances tumor cell-platelet binding and hematogenous metastasis. Hypoxia-induced upregulation of TF appears to parallel that of VEGF, although the mechanism remains unclear.

 

  • References

  • 1 Dvorak HF, Harvey VS, Estrella P, Brown LF, McDonagh J, Dvorak AM. Fibrin containing gels induce angiogenesis. Implications for tumor stroma generation and wound healing. Lab Invest 1987; 57: 673-86.
    PubMedGoogle Scholar
  • 2 Edwards RL, Silver J, Rickles FR. Human tumor procoagulants: Registry of the Subcommittee of Haemostasis of the Scientific and Standardization Committee, International Society on Thrombosis and Haemostasis. Thromb Haemost 1993; 69: 205-13.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Herbert JM, Savi P, Laplace MC, Lale A. IL4 inhibits LPSinduced, IL1-betai-nduced and TNFalpha-induced expression of tissue factor in endothelial cells and monocytes. FEBS Lett 1992; 310: 31-3.
    CrossrefPubMedGoogle Scholar
  • 4 Walz DA, Fenton JW. The role of thrombin in tumor cell metastasis. Invasion Metastasis 1994; 14: 303-8.
    PubMedGoogle Scholar
  • 5 Nagy JA, Brown LF, Senger DR, Lanir N, van de Water L, Dvorak AM, Dvorak HF. Pathogenesis of tumor stroma generation: a critical role for leaky blood vessels and fibrin deposition. Biochim Biophys Acta 1988; 948: 305-26.
    PubMedGoogle Scholar
  • 6 Qi J, Kreutzer D. Fibrin activation of vascular endothelial cells: induction of IL8. Journal Of Immunology 1995; 155: 867-76.
    PubMedGoogle Scholar
  • 7 Folkman J, Klagsbrun M. Angiogenic factors. Science 1987; 235: 442-7.
    CrossrefPubMedGoogle Scholar
  • 8 Zhang Y, Deng Y, Luther T, Müller M, Ziegler R, Waldherr R, Stern DM, Nawroth PP. Tissue factor controls the balance of angiogenic and antiangiogenic properties of tumor cells in mice. J Clin Invest 1994; 94: 1320-7.
    CrossrefPubMedGoogle Scholar
  • 9 Bromberg ME, Konigsberg WH, Madison J, Pawashe A, Garen A. Tissue factor promotes melanoma metastasis by a pathway independent of blood coagulation. Proc. Natl. Acad. Sci. USA 1995; 92: 8205-9.
    CrossrefPubMedGoogle Scholar
  • 10 Contrino J, Hair G, Kreutzer DL, Rickles FR. In situ detection of tissue factor in vascular endothelial cells: correlation with the malignant phenotype of human breast disease. Nat Med 1996; 2: 209-15.
    CrossrefPubMedGoogle Scholar
  • 11 Shoji M, Abe K, Nawroth PP, Rickles FR. Molecular mechanisms linking thrombosis and angiogenesis in cancer. Trends In Cardiovascular Medicine 1997; 7: 529.
    PubMedGoogle Scholar
  • 12 Ferrara N. Vascular Endothelial Growth Factor. Eur J Cancer 1996; 32A: 2413-22.
    PubMedGoogle Scholar
  • 13 Shimizu S. Induction of apoptosis as well as necrosis by hypoxia and predominant prevention of apoptosis by Bcl-2 and Bcl-X. Cancer Res 1996; 56: 2161-6.
    PubMedGoogle Scholar
  • 14 Shweioki D, Itin A, Soffer D, Keshet E. Vascular endothelial growth factor induced by hypoxia may mediate hypoxiainitiated angiogenesis. Nature 1992; 359: 843-5.
    CrossrefPubMedGoogle Scholar
  • 15 Herbert JM, Corseaux D, Lale A, Bernat A. Hypoxia primes endotoxininduced tissue factor expression in human monocytes and endothelial cells by a PAFDependent mechanism. J Cell Physiol 1996; 169: 290-9.
    CrossrefPubMedGoogle Scholar
  • 16 Ramani M, Khechai F, Ollivier V, Ternisien C, Bridey F, Hakim J, de Prost D. Interleukin-10 and pentoxifylline inhibit Creactive proteininduced tissue factor gene expression in peripheral human blood monocytes. FEBS Lett 1994; 356: 86-8.
    CrossrefPubMedGoogle Scholar
  • 17 Deprost D. Pentoxifylline: A potential treatment for thrombosis associated with abnormal tissue factor expression by monocytes and endothelial cells. J Cardiovasc Pharmacol 1995; 25: S114-118.
    CrossrefPubMedGoogle Scholar
  • 18 Amirkhosravi A, Biggerstaff JP, Warnes G, Francis DA, Francis JL. Determination of tumor cell procoagulant activity by Sonoclot (TM) analysis in whole blood. Thromb Res 1996; 84: 323-32.
    CrossrefPubMedGoogle Scholar
  • 19 Bradford M. A rapid and sensitive method for the quantitation of of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976; 72: 248-54.
    CrossrefPubMedGoogle Scholar
  • 20 Rickles FR, Levine M, Edwards RL. Hemostatic alterations in cancer patients. Cancer Metastasis Rev 1992; 11: 23748.
    PubMedGoogle Scholar
  • 21 Bick RL. Coagulation abnormalities in malignancy – a review. Semin Thromb Hemostasis 1992; 18: 353-72.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Zacharski LR, Memoli VA, Costantini V, Wojtukiewicz MZ, Ornstein DL. Clotting factors in tumour tissue: implications for cancer therapy. Blood Coagul Fibrinolysis 1990; 1: 718.
    PubMedGoogle Scholar
  • 23 Dvorak HF, Senger DR, Dvorak AM. Fibrin as a component of the tumour stroma: origins and biological significance. Cancer Metast Rev 1983; 2: 4173.
    PubMedGoogle Scholar
  • 24 Contrino J, Hair G, Kreutzer DL, Rickles FR. In situ detection of tissue factor in vascular endothelial cells: Correlation with the malignant phenotype of human breast disease. Nature Med 1996; 2: 209-15.
    CrossrefPubMedGoogle Scholar
  • 25 Yan SF, Gao Y, Zhai C, Oeth P, Mackman N. Hypoxia-induced expression of tissue factor is mediated by the Transcription Factor EGR1. Circulation 1997; 96: I662.
    PubMedGoogle Scholar
  • 26 Amirkhosravi M, Francis JL. Coagulation activation by MC28 fibrosarcoma cells facilitates lung tumor formation. Thromb Haemost 1995; 73: 59-65.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Leclerc NE, HaanArchipoff G, Lenoble M, Beretz A. Inhibitors of phosphodiesterase (pentoxifylline, trequinsin) inhibit apical and subcellular matrix expression of tissue factor in cultured human endothelial cells. J Cardiovasc Pharmacol 1995; 25: S88-91.
    CrossrefPubMedGoogle Scholar
  • 28 Su SJ, Chang KL, Lin TM, Huang YH, Yeh TM. Uromodulin and TammHorsfall protein induce human monocytes to secrete TNF and express tissue factor. Immunology 1997; 158: 3449-56.
    PubMedGoogle Scholar
  • 29 Amirkhosravi A, Warnes G, Biggerstaff J, Malik Z, May K, Francis JL. The effect of pentoxifylline on spontaneous and experimental metastasis of the mouse Neuro2a neuroblastoma. Clin Expl Metastasis 1997; 15: 453-61.
    CrossrefPubMedGoogle Scholar
  • 30 Melnyk O, Shuman MA, Kim KJ. Vascular Endothelial Growth Factor Promotes Tumors Dissemination by a Mechanism Distinct from Its Effect on Primary Tumor Growth. Cancer Res 1996; 56: 921-4.
    PubMedGoogle Scholar
  • 31 Mazure NM, Chen EY, Laderoute KR, and Giaccia AJ. Induction of Vascular Endothelial Growth Factor by Hypoxia Is Modulated by a Phosphatidylinositol 3-Kinase/Akt Signalling Pathway in Ha-ras-Transformed Cells Through a Hypoxia Inducible Factor-1 Transcriptional Element. Blood 1997; 90: 3322-31.
    PubMedGoogle Scholar
  • 32 Clauss M, Grell M, Fangmann C, Fiers W, Scheurich P, Risau W. Synergistic induction of endothelial tissue factor by tumor necrosis factor and vascular endothelial growth factor: Functional analysis of the tumor necrosis factor receptors. FEBS Lett 1996; 390: 3348.
    PubMedGoogle Scholar
  • 33 Ollivier V, Bentolila S, Chabbat J, Hakim J, and de Prost D. Tissue Factor-Dependent Vascular Endothelial Growth Factor Production by Human Fibroblasts in Response to Activated Factor VII. Blood 1998; 91: 2698-703.
    PubMedGoogle Scholar
  • 34 Mueller BM, and Ruf W. Requirement for binding of catalytically active factor VIIa in tissue factor-dependent experimental metastasis. J. Clin. Invest. 1998; 7: 1372-8.
    PubMedGoogle Scholar
  • 35 Pepper MS, Mandriota SJ, Vassalli JD, Orci L, Montesano R. Angiogenesis regulating cytokines: activities and interactions. Curr Top Microbiol Immunol 1996; 213: 31-67.
    PubMedGoogle Scholar
  • 36 Pepper MS. Manipulating angiogenesis – From basic science to the bedside. Arterioscler Thromb Vasc Biol. 1997; 17: 605-19.
    CrossrefPubMedGoogle Scholar
  • 37 Risau W. What, if anything, is an angiogenic factor?. Cancer Metast Rev 1996; 15: 149-51.
    CrossrefPubMedGoogle Scholar
  • 38 Clauss M, Gerlach M, Gerlach H, Brett J, Wang F, Familletti PC, Pan YCE, Olander JV, Connolly DT, Stern D. Vascularpermeability factor – a tumor-derived polypeptide that induces endothelialcell and monocyte procoagulant activity, and promotes monocyte migration. Experimental Medicine 1990; 172: 1535-45.
    CrossrefPubMedGoogle Scholar