Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1998 Feb;152(2):399–411.

Activation of coagulation and angiogenesis in cancer: immunohistochemical localization in situ of clotting proteins and vascular endothelial growth factor in human cancer.

M Shoji 1, W W Hancock 1, K Abe 1, C Micko 1, K A Casper 1, R M Baine 1, J N Wilcox 1, I Danave 1, D L Dillehay 1, E Matthews 1, J Contrino 1, J H Morrissey 1, S Gordon 1, T S Edgington 1, B Kudryk 1, D L Kreutzer 1, F R Rickles 1
PMCID: PMC1857968  PMID: 9466566

Abstract

Thrombin-catalyzed, cross-linked fibrin (XLF) formation is a characteristic histopathological finding in many human and experimental tumors and is thought to be of importance in the local host defense response. Although the pathogenesis of tumor-associated fibrin deposition is not entirely clear, several tumor procoagulants have been described as likely primary stimuli for the generation of thrombin (and XLF) in the tumor microenvironment (TME). In a previous study of a variety of human tumors we have shown that tissue factor (TF) is the major procoagulant. However, the relative contribution to fibrin deposition in the TME of tumor cell TF and host cell TF (eg, macrophage-derived) was not established. In addition, recent evidence has implicated TF in the regulation of the synthesis of the pro-angiogenic factor vascular endothelial growth factor (VEGF) by tumor cells. In the current study we used in situ techniques to determine the cellular localization of XLF, TF, VEGF, and an alternative tumor procoagulant, so-called cancer procoagulant (CP), a cysteine protease that activates clotting factor X. In lung cancer we have found XLF localized predominantly to the surface of tumor-associated macrophages, as well as to some endothelial cells and perivascular fibroblasts in the stromal area of the tumors co-distributed with TF at the interface of the tumor and host cells. Cancer pro-coagulant was localized to tumor cells in several cases but not in conjunction with the deposition of XLF. TF and VEGF were co-localized in both lung cancer and breast cancer cells by in situ hybridization and immunohistochemical staining. Furthermore, a strong relationship was found between the synthesis of TF and VEGF levels in human breast cancer cell lines (r2 = 0.84; P < 0.0001). Taken together, these data are consistent with a highly complex interaction between tumor cells, macrophages, and endothelial cells in the TME leading to fibrin formation and tumor angiogenesis.

Full text

PDF
399

Images in this article

404Figure 1
on p.404
405Figure 2
on p.405
406Figure 3
on p.406
407Figure 4
on p.407
408Figure 5
on p.408

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alessio M. G., Falanga A., Consonni R., Bassan R., Minetti B., Donati M. B., Barbui T. Cancer procoagulant in acute lymphoblastic leukemia. Eur J Haematol. 1990 Aug;45(2):78–81. doi: 10.1111/j.1600-0609.1990.tb00421.x. [DOI] [PubMed] [Google Scholar]
  2. Brett J., Gerlach H., Nawroth P., Steinberg S., Godman G., Stern D. Tumor necrosis factor/cachectin increases permeability of endothelial cell monolayers by a mechanism involving regulatory G proteins. J Exp Med. 1989 Jun 1;169(6):1977–1991. doi: 10.1084/jem.169.6.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clauss M., Gerlach M., Gerlach H., Brett J., Wang F., Familletti P. C., Pan Y. C., Olander J. V., Connolly D. T., Stern D. Vascular permeability factor: a tumor-derived polypeptide that induces endothelial cell and monocyte procoagulant activity, and promotes monocyte migration. J Exp Med. 1990 Dec 1;172(6):1535–1545. doi: 10.1084/jem.172.6.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Contrino J., Goralnick S., Qi J., Hair G., Rickles F. R., Kreutzer D. L. Fibrin induction of tissue factor expression in human vascular endothelial cells. Circulation. 1997 Jul 15;96(2):605–613. [PubMed] [Google Scholar]
  5. Contrino J., Hair G. A., Schmeizl M. A., Rickles F. R., Kreutzer D. L. In situ characterization of antigenic and functional tissue factor expression in human tumors utilizing monoclonal antibodies and recombinant factor VIIa as probes. Am J Pathol. 1994 Dec;145(6):1315–1322. [PMC free article] [PubMed] [Google Scholar]
  6. Contrino J., Hair G., Kreutzer D. L., Rickles F. R. In situ detection of tissue factor in vascular endothelial cells: correlation with the malignant phenotype of human breast disease. Nat Med. 1996 Feb;2(2):209–215. doi: 10.1038/nm0296-209. [DOI] [PubMed] [Google Scholar]
  7. Donati M. B., Gambacorti-Passerini C., Casali B., Falanga A., Vannotti P., Fossati G., Semeraro N., Gordon S. G. Cancer procoagulant in human tumor cells: evidence from melanoma patients. Cancer Res. 1986 Dec;46(12 Pt 1):6471–6474. [PubMed] [Google Scholar]
  8. Drake T. A., Cheng J., Chang A., Taylor F. B., Jr Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol. 1993 May;142(5):1458–1470. [PMC free article] [PubMed] [Google Scholar]
  9. Drake T. A., Morrissey J. H., Edgington T. S. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol. 1989 May;134(5):1087–1097. [PMC free article] [PubMed] [Google Scholar]
  10. Dvorak H. F., Brown L. F., Detmar M., Dvorak A. M. Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol. 1995 May;146(5):1029–1039. [PMC free article] [PubMed] [Google Scholar]
  11. Edwards R. L., Morgan D. L., Rickles F. R. Animal tumor procoagulants: registry of the Subcommittee on Haemostasis and Malignancy of the Scientific and Standardization Committee, International Society of Thrombosis and Haemostasis. Thromb Haemost. 1990 Feb 19;63(1):133–138. [PubMed] [Google Scholar]
  12. Edwards R. L., Rickles F. R., Bobrove A. M. Mononuclear cell tissue factor: cell of origin and requirements for activation. Blood. 1979 Aug;54(2):359–370. [PubMed] [Google Scholar]
  13. Edwards R. L., Rickles F. R., Cronlund M. Abnormalities of blood coagulation in patients with cancer. Mononuclear cell tissue factor generation. J Lab Clin Med. 1981 Dec;98(6):917–928. [PubMed] [Google Scholar]
  14. Edwards R. L., Silver J., Rickles F. R. Human tumor procoagulants: registry of the Subcommittee on Haemostasis and Malignancy of the Scientific and Standardization Committee, International Society on Thrombosis and Haemostasis. Thromb Haemost. 1993 Feb 1;69(2):205–213. [PubMed] [Google Scholar]
  15. Ewan V. A., Cieplinski W., Hancock W. W., Goldschneider I., Boyd A. W., Rickles F. R. Production and characterization of a monoclonal antibody (A1-3) that binds selectively to activated monocytes and inhibits monocyte procoagulant activity. J Immunol. 1986 Apr 1;136(7):2408–2415. [PubMed] [Google Scholar]
  16. Falanga A., Alessio M. G., Donati M. B., Barbui T. A new procoagulant in acute leukemia. Blood. 1988 Apr;71(4):870–875. [PubMed] [Google Scholar]
  17. Folkman J. Tumor angiogenesis and tissue factor. Nat Med. 1996 Feb;2(2):167–168. doi: 10.1038/nm0296-167. [DOI] [PubMed] [Google Scholar]
  18. Gordon S. G., Cross B. A. An enzyme-linked immunosorbent assay for cancer procoagulant and its potential as a new tumor marker. Cancer Res. 1990 Oct 1;50(19):6229–6234. [PubMed] [Google Scholar]
  19. Gregory S. A., Edgington T. S. Tissue factor induction in human monocytes. Two distinct mechanisms displayed by different alloantigen-responsive T cell clones. J Clin Invest. 1985 Dec;76(6):2440–2445. doi: 10.1172/JCI112260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gregory S. A., Kornbluth R. S., Helin H., Remold H. G., Edgington T. S. Monocyte procoagulant inducing factor: a lymphokine involved in the T cell-instructed monocyte procoagulant response to antigen. J Immunol. 1986 Nov 15;137(10):3231–3239. [PubMed] [Google Scholar]
  21. Gregory S. A., Morrissey J. H., Edgington T. S. Regulation of tissue factor gene expression in the monocyte procoagulant response to endotoxin. Mol Cell Biol. 1989 Jun;9(6):2752–2755. doi: 10.1128/mcb.9.6.2752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grignani G., Falanga A., Pacchiarini L., Alessio M. G., Zucchella M., Fratino P., Donati M. B. Human breast and colon carcinomas express cysteine proteinase activities with pro-aggregating and pro-coagulant properties. Int J Cancer. 1988 Oct 15;42(4):554–557. doi: 10.1002/ijc.2910420414. [DOI] [PubMed] [Google Scholar]
  23. Hair G. A., Padula S., Zeff R., Schmeizl M., Contrino J., Kreutzer D. L., de Moerloose P., Boyd A. W., Stanley I., Burgess A. W. Tissue factor expression in human leukemic cells. Leuk Res. 1996 Jan;20(1):1–11. doi: 10.1016/0145-2126(95)00107-7. [DOI] [PubMed] [Google Scholar]
  24. Hancock W. W., Atkins R. C. Immunohistological studies with monoclonal antibodies. Methods Enzymol. 1986;121:828–848. doi: 10.1016/0076-6879(86)21080-0. [DOI] [PubMed] [Google Scholar]
  25. Hancock W. W., Gee D., De Moerloose P., Rickles F. R., Ewan V. A., Atkins R. C. Immunohistological analysis of serial biopsies taken during human renal allograft rejection. Changing profile of infiltrating cells and activation of the coagulation system. Transplantation. 1985 Apr;39(4):430–438. doi: 10.1097/00007890-198504000-00018. [DOI] [PubMed] [Google Scholar]
  26. Hancock W. W., Rickles F. R., Ewan V. A., Atkins R. C. Immunohistological studies with A1-3, a monoclonal antibody to activated human monocytes and macrophages. J Immunol. 1986 Apr 1;136(7):2416–2420. [PubMed] [Google Scholar]
  27. Hogg N. Human monocytes are associated with the formation of fibrin. J Exp Med. 1983 Feb 1;157(2):473–485. doi: 10.1084/jem.157.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kudryk B., Rohoza A., Ahadi M., Chin J., Wiebe M. E. A monoclonal antibody with ability to distinguish between NH2-terminal fragments derived from fibrinogen and fibrin. Mol Immunol. 1983 Nov;20(11):1191–1200. doi: 10.1016/0161-5890(83)90142-6. [DOI] [PubMed] [Google Scholar]
  29. Kudryk B., Rohoza A., Ahadi M., Chin J., Wiebe M. E. Specificity of a monoclonal antibody for the NH2-terminal region of fibrin. Mol Immunol. 1984 Jan;21(1):89–94. doi: 10.1016/0161-5890(84)90093-2. [DOI] [PubMed] [Google Scholar]
  30. Lorenzet R., Peri G., Locati D., Allavena P., Colucci M., Semeraro N., Mantovani A., Donati M. B. Generation of procoagulant activity by mononuclear phagocytes: a possible mechanism contributing to blood clotting activation within malignant tissues. Blood. 1983 Aug;62(2):271–273. [PubMed] [Google Scholar]
  31. Mann K. G., Nesheim M. E., Church W. R., Haley P., Krishnaswamy S. Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood. 1990 Jul 1;76(1):1–16. [PubMed] [Google Scholar]
  32. Morgan D., Edwards R. L., Rickles F. R. Monocyte procoagulant activity as a peripheral marker of clotting activation in cancer patients. Haemostasis. 1988;18(1):55–65. doi: 10.1159/000215783. [DOI] [PubMed] [Google Scholar]
  33. Morrissey J. H., Fair D. S., Edgington T. S. Monoclonal antibody analysis of purified and cell-associated tissue factor. Thromb Res. 1988 Nov 1;52(3):247–261. doi: 10.1016/0049-3848(88)90084-9. [DOI] [PubMed] [Google Scholar]
  34. Nagy J. A., Brown L. F., Senger D. R., Lanir N., Van de Water L., Dvorak A. M., Dvorak H. F. Pathogenesis of tumor stroma generation: a critical role for leaky blood vessels and fibrin deposition. Biochim Biophys Acta. 1989 Feb;948(3):305–326. doi: 10.1016/0304-419x(89)90004-8. [DOI] [PubMed] [Google Scholar]
  35. O'MEARA R. A. Coagulative properties of cancers. Ir J Med Sci. 1958 Oct;394:474–479. [PubMed] [Google Scholar]
  36. Qi J., Kreutzer D. L. Fibrin activation of vascular endothelial cells. Induction of IL-8 expression. J Immunol. 1995 Jul 15;155(2):867–876. [PubMed] [Google Scholar]
  37. Rickles F. R., Edwards R. L. Activation of blood coagulation in cancer: Trousseau's syndrome revisited. Blood. 1983 Jul;62(1):14–31. [PubMed] [Google Scholar]
  38. Rickles F. R., Hancock W. W. Antimetastatic agents. II. Summary of the interactions of tumor cells with blood coagulation factors, platelets, fibrinolytic factors, and inflammatory cells and their soluble mediators: potential for therapeutic interventions. Semin Thromb Hemost. 1988 Jan;14(1):126–132. doi: 10.1055/s-2007-1002765. [DOI] [PubMed] [Google Scholar]
  39. Rickles F. R., Hancock W. W., Edwards R. L., Zacharski L. R. Antimetastatic agents. I. Role of cellular procoagulants in the pathogenesis of fibrin deposition in cancer and the use of anticoagulants and/or antiplatelet drugs in cancer treatment. Semin Thromb Hemost. 1988 Jan;14(1):88–94. doi: 10.1055/s-2007-1002760. [DOI] [PubMed] [Google Scholar]
  40. Rickles F. R., Levine M., Edwards R. L. Hemostatic alterations in cancer patients. Cancer Metastasis Rev. 1992 Nov;11(3-4):237–248. doi: 10.1007/BF01307180. [DOI] [PubMed] [Google Scholar]
  41. Ruf W., Edgington T. S. An anti-tissue factor monoclonal antibody which inhibits TF.VIIa complex is a potent anticoagulant in plasma. Thromb Haemost. 1991 Nov 1;66(5):529–533. [PubMed] [Google Scholar]
  42. Semeraro N., De Lucia O., Lattanzio A., Montemurro P., Giordano D., Loizzi M., Carpagnano F. Procoagulant activity of human alveolar macrophages: different expression in patients with lung cancer. Int J Cancer. 1986 Apr 15;37(4):525–529. doi: 10.1002/ijc.2910370409. [DOI] [PubMed] [Google Scholar]
  43. Semeraro N., Montemurro P., Conese M., Giordano D., Stella M., Restaino A., Cagnazzo G., Colucci M. Procoagulant activity of mononuclear phagocytes from different anatomical sites in patients with gynecological malignancies. Int J Cancer. 1990 Feb 15;45(2):251–254. doi: 10.1002/ijc.2910450208. [DOI] [PubMed] [Google Scholar]
  44. Senger D. R., Galli S. J., Dvorak A. M., Perruzzi C. A., Harvey V. S., Dvorak H. F. Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science. 1983 Feb 25;219(4587):983–985. doi: 10.1126/science.6823562. [DOI] [PubMed] [Google Scholar]
  45. VanDeWater L., Tracy P. B., Aronson D., Mann K. G., Dvorak H. F. Tumor cell generation of thrombin via functional prothrombinase assembly. Cancer Res. 1985 Nov;45(11 Pt 1):5521–5525. [PubMed] [Google Scholar]
  46. Walz D. A., Fenton J. W. The role of thrombin in tumor cell metastasis. Invasion Metastasis. 1994;14(1-6):303–308. [PubMed] [Google Scholar]
  47. Wilcox J. N. Fundamental principles of in situ hybridization. J Histochem Cytochem. 1993 Dec;41(12):1725–1733. doi: 10.1177/41.12.8245419. [DOI] [PubMed] [Google Scholar]
  48. Wilcox J. N., Smith K. M., Schwartz S. M., Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2839–2843. doi: 10.1073/pnas.86.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wilcox J. N., Smith K. M., Williams L. T., Schwartz S. M., Gordon D. Platelet-derived growth factor mRNA detection in human atherosclerotic plaques by in situ hybridization. J Clin Invest. 1988 Sep;82(3):1134–1143. doi: 10.1172/JCI113671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wojtukiewicz M. Z., Zacharski L. R., Memoli V. A., Kisiel W., Kudryk B. J., Rousseau S. M., Stump D. C. Abnormal regulation of coagulation/fibrinolysis in small cell carcinoma of the lung. Cancer. 1990 Feb 1;65(3):481–485. doi: 10.1002/1097-0142(19900201)65:3<481::aid-cncr2820650318>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  51. Zhang Y., Deng Y., Luther T., Müller M., Ziegler R., Waldherr R., Stern D. M., Nawroth P. P. Tissue factor controls the balance of angiogenic and antiangiogenic properties of tumor cells in mice. J Clin Invest. 1994 Sep;94(3):1320–1327. doi: 10.1172/JCI117451. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES