Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1996 Apr;73(7):902–908. doi: 10.1038/bjc.1996.161

Thrombin enhances the adhesion and migration of human colon adenocarcinoma cells via increased beta 3-integrin expression on the tumour cell surface and their inhibition by the snake venom peptide, rhodostomin.

H S Chiang 1, R S Yang 1, T F Huang 1
PMCID: PMC2074261  PMID: 8611404

Abstract

The interactions between tumour cells and the microvasculature, including the adhesion of tumour cells to endothelium and extracellular matrix (ECM) as well as their migratory ability, are prerequisites for metastasis to occur. In this study we showed that thrombin is capable of enhancing in vitro tumour cell metastatic potential in terms of adhesive properties and migratory response. Following exposure to subclotting concentrations of thrombin, SW-480 human colon adenocarcinoma cells exhibited increased adhesion to both the endothelium and ECM component (i.e. fibronectin). Likewise, the pretreatment of thrombin enhanced the migratory ability of SW-480 cells. The enhanced adhesion was significantly inhibited by complexing of thrombin with its inhibitor hirudin, or by serine proteinase inhibition with 3,4-DCI, but was unaffected by pretreatment of tumour cells with actinomycin D or cycloheximide. The effect of thrombin resulted in an upregulated cell-surface expression of beta 3 integrins, a group of receptors mediating interactions between tumour cells and endothelial cells, and between tumour cells and ECM. Antibodies against beta 3 integrins effectively blocked both the enhanced adhesion and migration. This thrombin-mediated up-regulation of beta 3 integrins involved the activation of protein kinase C (PKC) as thrombin-enhanced adhesion was diminished by PKC inhibition. Rhodostomin, an Arg-Gly-Asp-containing antiplatelet snake venom peptide that antagonises the binding of ECM toward beta 3 integrins on SW-480 cells, was about 600 and 500 times, more potent that RGDS in inhibiting thrombin-enhanced adhesion and migration respectively. Our data suggest that PKC inhibitors as well as rhodostomin may serve as inhibitory agents in the prevention of thrombin-enhanced metastasis.

Full text

PDF
908

Selected References

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

  1. Brown P. J. Phorbol ester stimulation of fibronectin-mediated cell adhesion. Biochem Biophys Res Commun. 1988 Sep 15;155(2):603–607. doi: 10.1016/s0006-291x(88)80537-0. [DOI] [PubMed] [Google Scholar]
  2. Bruhn H. D., Zurborn K. H. Influences of clotting factors (thrombin, factor XIII) and of fibronectin on the growth of tumor cells and leukemic cells in vitro. Blut. 1983 Feb;46(2):85–88. doi: 10.1007/BF00320663. [DOI] [PubMed] [Google Scholar]
  3. Cavanaugh P. G., Sloane B. F., Honn K. V. Role of the coagulation system in tumor-cell-induced platelet aggregation and metastasis. Haemostasis. 1988;18(1):37–46. doi: 10.1159/000215781. [DOI] [PubMed] [Google Scholar]
  4. Chiang H. S., Peng H. C., Huang T. F. Characterization of integrin expression and regulation on SW-480 human colon adenocarcinoma cells and the effect of rhodostomin on basal and upregulated tumor cell adhesion. Biochim Biophys Acta. 1994 Dec 30;1224(3):506–516. doi: 10.1016/0167-4889(94)90288-7. [DOI] [PubMed] [Google Scholar]
  5. Chiang H. S., Swaim M. W., Huang T. F. Characterization of platelet aggregation induced by human colon adenocarcinoma cells and its inhibition by snake venom peptides, trigramin and rhodostomin. Br J Haematol. 1994 Jun;87(2):325–331. doi: 10.1111/j.1365-2141.1994.tb04917.x. [DOI] [PubMed] [Google Scholar]
  6. Chopra H., Hatfield J. S., Chang Y. S., Grossi I. M., Fitzgerald L. A., O'Gara C. Y., Marnett L. J., Diglio C. A., Taylor J. D., Honn K. V. Role of tumor cytoskeleton and membrane glycoprotein IRGpIIb/IIIa in platelet adhesion to tumor cell membrane and tumor cell-induced platelet aggregation. Cancer Res. 1988 Jul 1;48(13):3787–3800. [PubMed] [Google Scholar]
  7. Chopra H., Timar J., Chen Y. Q., Rong X. H., Grossi I. M., Fitzgerald L. A., Taylor J. D., Honn K. V. The lipoxygenase metabolite 12(S)-HETE induces a cytoskeleton-dependent increase in surface expression of integrin alpha IIb beta 3 on melanoma cells. Int J Cancer. 1991 Nov 11;49(5):774–786. doi: 10.1002/ijc.2910490524. [DOI] [PubMed] [Google Scholar]
  8. Costantini V., Zacharski L. R. Fibrin and cancer. Thromb Haemost. 1993 May 3;69(5):406–414. [PubMed] [Google Scholar]
  9. DiCorleto P. E., de la Motte C. A. Thrombin causes increased monocytic-cell adhesion to endothelial cells through a protein kinase C-dependent pathway. Biochem J. 1989 Nov 15;264(1):71–77. doi: 10.1042/bj2640071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Du X., Gu M., Weisel J. W., Nagaswami C., Bennett J. S., Bowditch R., Ginsberg M. H. Long range propagation of conformational changes in integrin alpha IIb beta 3. J Biol Chem. 1993 Nov 5;268(31):23087–23092. [PubMed] [Google Scholar]
  11. Fenton J. W., 2nd Regulation of thrombin generation and functions. Semin Thromb Hemost. 1988 Jul;14(3):234–240. doi: 10.1055/s-2007-1002783. [DOI] [PubMed] [Google Scholar]
  12. Fitzgerald L. A., Phillips D. R. Calcium regulation of the platelet membrane glycoprotein IIb-IIIa complex. J Biol Chem. 1985 Sep 15;260(20):11366–11374. [PubMed] [Google Scholar]
  13. Ginsberg M. H., Du X., Plow E. F. Inside-out integrin signalling. Curr Opin Cell Biol. 1992 Oct;4(5):766–771. doi: 10.1016/0955-0674(92)90099-x. [DOI] [PubMed] [Google Scholar]
  14. Gomez M. L., Medrano E. E., Cafferatta E. G., Tellez-Inon M. T. Protein kinase C is differentially regulated by thrombin, insulin, and epidermal growth factor in human mammary tumor cells. Exp Cell Res. 1988 Mar;175(1):74–80. doi: 10.1016/0014-4827(88)90256-x. [DOI] [PubMed] [Google Scholar]
  15. Greco N. J., Jamieson G. A. High and moderate affinity pathways for alpha-thrombin-induced platelet activation. Proc Soc Exp Biol Med. 1991 Dec;198(3):792–799. doi: 10.3181/00379727-198-43321d. [DOI] [PubMed] [Google Scholar]
  16. Grossi I. M., Fitzgerald L. A., Umbarger L. A., Nelson K. K., Diglio C. A., Taylor J. D., Honn K. V. Bidirectional control of membrane expression and/or activation of the tumor cell IRGpIIb/IIIa receptor and tumor cell adhesion by lipoxygenase products of arachidonic acid and linoleic acid. Cancer Res. 1989 Feb 15;49(4):1029–1037. [PubMed] [Google Scholar]
  17. Grossi I. M., Hatfield J. S., Fitzgerald L. A., Newcombe M., Taylor J. D., Honn K. V. Role of tumor cell glycoproteins immunologically related to glycoproteins Ib and IIb/IIIa in tumor cell-platelet and tumor cell-matrix interactions. FASEB J. 1988 May;2(8):2385–2395. doi: 10.1096/fasebj.2.8.2452113. [DOI] [PubMed] [Google Scholar]
  18. Honn K. V., Chen Y. Q., Timar J., Onoda J. M., Hatfield J. S., Fligiel S. E., Steinert B. W., Diglio C. A., Grossi I. M., Nelson K. K. Alpha IIb beta 3 integrin expression and function in subpopulations of murine tumors. Exp Cell Res. 1992 Jul;201(1):23–32. doi: 10.1016/0014-4827(92)90344-8. [DOI] [PubMed] [Google Scholar]
  19. Huang T. F., Holt J. C., Kirby E. P., Niewiarowski S. Trigramin: primary structure and its inhibition of von Willebrand factor binding to glycoprotein IIb/IIIa complex on human platelets. Biochemistry. 1989 Jan 24;28(2):661–666. doi: 10.1021/bi00428a037. [DOI] [PubMed] [Google Scholar]
  20. Huang T. F., Holt J. C., Lukasiewicz H., Niewiarowski S. Trigramin. A low molecular weight peptide inhibiting fibrinogen interaction with platelet receptors expressed on glycoprotein IIb-IIIa complex. J Biol Chem. 1987 Nov 25;262(33):16157–16163. [PubMed] [Google Scholar]
  21. Huang T. F., Liu C. Z., Ouyang C. H., Teng C. M. Halysin, an antiplatelet Arg-Gly-Asp-containing snake venom peptide, as fibrinogen receptor antagonist. Biochem Pharmacol. 1991 Aug 22;42(6):1209–1219. doi: 10.1016/0006-2952(91)90256-5. [DOI] [PubMed] [Google Scholar]
  22. Huang T. F., Wang W. J., Teng C. M., Ouyang C. Mechanism of action of the antiplatelet peptide, arietin, from Bitis arietans venom. Biochim Biophys Acta. 1991 May 24;1074(1):144–150. doi: 10.1016/0304-4165(91)90053-j. [DOI] [PubMed] [Google Scholar]
  23. Huang T. F., Wu Y. J., Ouyang C. Characterization of a potent platelet aggregation inhibitor from Agkistrodon rhodostoma snake venom. Biochim Biophys Acta. 1987 Sep 11;925(3):248–257. doi: 10.1016/0304-4165(87)90189-9. [DOI] [PubMed] [Google Scholar]
  24. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liu B., Renaud C., Nelson K. K., Chen Y. Q., Bazaz R., Kowynia J., Timar J., Diglio C. A., Honn K. V. Protein-kinase-C inhibitor calphostin C reduces B16 amelanotic melanoma cell adhesion to endothelium and lung colonization. Int J Cancer. 1992 Aug 19;52(1):147–152. doi: 10.1002/ijc.2910520126. [DOI] [PubMed] [Google Scholar]
  26. Manning D. R., Brass L. F. The role of GTP-binding proteins in platelet activation. Thromb Haemost. 1991 Oct 1;66(4):393–399. [PubMed] [Google Scholar]
  27. McGregor B. C., McGregor J. L., Weiss L. M., Wood G. S., Hu C. H., Boukerche H., Warnke R. A. Presence of cytoadhesins (IIb-IIIa-like glycoproteins) on human metastatic melanomas but not on benign melanocytes. Am J Clin Pathol. 1989 Oct;92(4):495–499. doi: 10.1093/ajcp/92.4.495. [DOI] [PubMed] [Google Scholar]
  28. Medrano E. E., Cafferata E. G., Larcher F. Role of thrombin in the proliferative response of T-47D mammary tumor cells. Mitogenic action and pleiotropic modifications induced together with epidermal growth factor and insulin. Exp Cell Res. 1987 Oct;172(2):354–364. doi: 10.1016/0014-4827(87)90393-4. [DOI] [PubMed] [Google Scholar]
  29. Nierodzik M. L., 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 Jun 15;52(12):3267–3272. [PubMed] [Google Scholar]
  30. O'Toole T. E., Mandelman D., Forsyth J., Shattil S. J., Plow E. F., Ginsberg M. H. Modulation of the affinity of integrin alpha IIb beta 3 (GPIIb-IIIa) by the cytoplasmic domain of alpha IIb. Science. 1991 Nov 8;254(5033):845–847. doi: 10.1126/science.1948065. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Rucinski B., Niewiarowski S., Holt J. C., Soszka T., Knudsen K. A. Batroxostatin, an Arg-Gly-Asp-containing peptide from Bothrops atrox, is a potent inhibitor of platelet aggregation and cell interaction with fibronectin. Biochim Biophys Acta. 1990 Sep 24;1054(3):257–262. doi: 10.1016/0167-4889(90)90096-v. [DOI] [PubMed] [Google Scholar]
  33. Shebuski R. J., Ramjit D. R., Bencen G. H., Polokoff M. A. Characterization and platelet inhibitory activity of bitistatin, a potent arginine-glycine-aspartic acid-containing peptide from the venom of the viper Bitis arietans. J Biol Chem. 1989 Dec 25;264(36):21550–21556. [PubMed] [Google Scholar]
  34. Vaporciyan A. A., Jones M. L., Ward P. A. Rapid analysis of leukocyte-endothelial adhesion. J Immunol Methods. 1993 Feb 26;159(1-2):93–100. doi: 10.1016/0022-1759(93)90145-w. [DOI] [PubMed] [Google Scholar]
  35. Vu T. K., Hung D. T., Wheaton V. I., Coughlin S. R. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell. 1991 Mar 22;64(6):1057–1068. doi: 10.1016/0092-8674(91)90261-v. [DOI] [PubMed] [Google Scholar]
  36. Wojtukiewicz M. Z., Tang D. G., Ben-Josef E., Renaud C., Walz D. A., Honn K. V. Solid tumor cells express functional "tethered ligand" thrombin receptor. Cancer Res. 1995 Feb 1;55(3):698–704. [PubMed] [Google Scholar]
  37. Wojtukiewicz M. Z., Tang D. G., Ciarelli J. J., Nelson K. K., Walz D. A., Diglio C. A., Mammen E. F., Honn K. V. Thrombin increases the metastatic potential of tumor cells. Int J Cancer. 1993 Jul 9;54(5):793–806. doi: 10.1002/ijc.2910540514. [DOI] [PubMed] [Google Scholar]
  38. Wojtukiewicz M. Z., Tang D. G., Nelson K. K., Walz D. A., Diglio C. A., Honn K. V. Thrombin enhances tumor cell adhesive and metastatic properties via increased alpha IIb beta 3 expression on the cell surface. Thromb Res. 1992 Nov 1;68(3):233–245. doi: 10.1016/0049-3848(92)90081-k. [DOI] [PubMed] [Google Scholar]
  39. Zacharski L. R., Costantini V., Wojtukiewicz M. Z., Memoli V. A., Kudryk B. J. Anticoagulants as cancer therapy. Semin Oncol. 1990 Apr;17(2):217–227. [PubMed] [Google Scholar]
  40. Zacharski L. R., Meehan K. R., Algarra S. M., Calvo F. A. Clinical trials with anticoagulant and antiplatelet therapies. Cancer Metastasis Rev. 1992 Nov;11(3-4):421–431. doi: 10.1007/BF01307191. [DOI] [PubMed] [Google Scholar]
  41. Zacharski L. R., Wojtukiewicz M. Z., Costantini V., Ornstein D. L., Memoli V. A. Pathways of coagulation/fibrinolysis activation in malignancy. Semin Thromb Hemost. 1992 Jan;18(1):104–116. doi: 10.1055/s-2007-1002415. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES