Summary
Platelets and coagulation are involved in the pathogenesis of blood-borne metastases. The aim of this study is to obtain more information about the mechanisms involved in the initial adhesion of tumor cells to endothelial cells. In short term experiments with tumor cells, suspended in the medium of cultured endothelial cells, we tested whether addition of both platelets and thrombin cause more tumor cell adhesion to endothelial cells, than when either platelets or thrombin are acting alone.
HeLa cells or HT29 cells, prelabeled with radioactive 51Cr, human platelets, and thrombin were added to human endothelial cell cultures. Following 15 min of shaking at 37° C, the percentage of tumor cell adhesion was calculated. The percentages of adhering tumor cells with the presence of both platelets and thrombin were greatly increased compared to controls. Addition of hirudin 2 min before thrombin lowered the adhesion percentage of tumor cells. Hirudin added immediately before and 2 min after thrombin gave only minor effects. When the endothelium was treated with superoxide dismutase, catalase, and mannitol, the adhesion of tumor cells was lowered with catalase and superoxide dismutase. The cause of tumor cell-endothelial cell interaction is probably complex. Our results show that activated platelets enhance the tumor cell adhesion, and that generation of active oxygen species may be important in the initial phase of the interaction.
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Abecassis, J.; Millon-Collard, R.; Klein-Soyer, C., et al. Adhesion of human breast cancer cell line MCF-7 to human vascular endothelial cells in culture: enhancement by activated platelets. Int. J. Cancer. 40:525–531; 1987.
Al-Mondhiry, H.; McGarvey, V. Tumor interaction with vascular endothelium. Haemostasis 17:245–253; 1987.
Bastida, E.; Escolar, G.; Ordinas, A., et al. Morphometric evaluation of thrombogenesis by microvesicles from human tumor cell lines with thrombin-dependent (U87MG) and adenosine diphosphate-dependent (SKNMC) platelet-activating mechanisms. J. Lab. Clin. Med. 108:622–627; 1986.
Bastida, E.; Ordinas, A. Platelet contribution to the formation of metastatic foci: the role of cancer cell-induced platelet activation. Haemostasis. 18:29–36; 1988.
Bastida, E.; Ordinas, A.; Giardina, S. L., et al. Differentiation of platelet-aggregating effects of human tumor cell lines based on inhibition studies with apyrase, hirudin, and phospholipase. Cancer Res. 42:4348–4352; 1982.
Bereta, M.; Bereta, J.; Cohen, S., et al. Effect of inflammatory cytokines on the adherence of tumor cell to endothelium in a murine model. Cell. Immunol. 136:263–277; 1991.
Boogaerts, M. A.; Van de Broeck, J.; Deckmyn, H., et al. Protective effect of vitamin E on immune triggered, granulocyte mediated endothelial injury. Thromb. Haemostasis 51:89–92; 1984.
Boukerche, H.; Berthier-Vergnes, O.; Bailly, M., et al. A monoclonal antibody (LYP18) directed against the blood platelet glycoprotein IIb/IIIa complex inhibits human melanoma growth in vivo. Blood. 74:909–912; 1989.
Burrows, F. J.; Haskard, D. O.; Hart, I. R., et al. Influence of tumorderived interleukin I on melanoma-endothelial cell interactions in vitro. Cancer Res. 51:4768–4775; 1991.
Crissman, J. D.; Hatfield, J. S.; Menter, D. G., et al. Morphological study of the interaction of intravascular tumor cells with endothelial cells and subendothelial matrix. Cancer Res. 48:4065–4072; 1988.
Crissman, J. D.; Hatfield, J.; Schaldenbrand, M., et al. Arrest and extravasation of B16 amelanotic melanoma in murine lungs: a light and electron microscopic study. Lab. Invest. 53:470–478; 1985.
Fong, K. L.; McCay, P. B.; Poyer, J. L., et al. Evidence that peroxidation of lysosomal membranes is initiated by hydroxyl free radicals produced during flavine enzyme activity. J. Biol. Chem. 248:7792–7797; 1973.
Gasic, G. J.; Gasic, T. B.; Galanti, N., et al. Platelet-tumor-cell interactions in mice. The role of platelets in the spread of malignant disease. Int. J. Cancer. 1:704–718; 1973.
Gasic, G. J.; Gasic, T. B.; Stewart, C. C. Antimetastatic effects associated with platelet reduction. Proc. Natl. Acad. Sci. USA 61:46–52; 1968.
Gastpar, H. Inhibition of cancer cell stickiness by the blocking of platelet aggregation. S. Afr. Med. J. 48:621–627; 1974.
Hara, Y.; Steiner, M.; Baldini, M. G. Characterization of the platelet-aggregating activity of tumor cells. Cancer Res. 40:1217–1222; 1980.
Helland, I.; Jørgensen, L. Interaction in vitro between HeLa-cells, thrombin-stimulated platelets, and endothelial cells or subendothelium. XII Congress International Society of Thrombosis and Haemostasis, Tokyo, Japan, 1989. Thromb. Haemostas 62:138; 1989.
Honn, K. V.; Busse, W. D.; Sloane, B. F. Prostacyclin and thromboxanes: implication for their role in tumor cell metastasis. Biochem. Pharmacol. 32:I-II; 1983.
Honn, K. V.; Grossi, I. M.; Diglio, C. A., et al. Enhanced tumor cell adhesion to the subendothelial matrix resulting from 12(S)-HETE-induced endothelial cell retraction. FASEB J. 3:2285–2293; 1989.
Honn, K. V.; Grossi, I. M.; Fitzgerald, L. A., et al. Lipoxygenase products regulate IRGpIIb/IIIa receptor mediated adhesion of tumor cells to endothelial cells, subendothelial matrix and fibronectin. Proc. Soc. Exp. Biol. Med. 189:130–135; 1988.
Honn, K. V.; Tang, D. G. Adhesion molecules and tumor cell interaction with endothelium and subendothelial matrix. Cancer Metastasis Rev. 11:353–375; 1992.
Jaffe, E. A.; Nachman, R. L.; Becker, C. G., et al. Culture of human endothelial cell derived from umbilical veins: identification by morphologic and immunologic criteria. J. Clin. Invest. 52:2745–2756; 1973.
Jørgensen, L.; Grøthe, A. G.; Larsen, T., et al. Injury to cultured endothelial cells by thrombin-stimulated platelets. Lab. Invest. 54:408–415; 1986.
Klepfish, A.; Greco, M. A.; Karpatkin, S. Thrombin stimulates melanoma tumor-cell binding to endothelial cells and subendothelial matrix. Int. J. Cancer. 53:978–982; 1993.
Kramer, R. H.; Enenstein, J.; Ramos, D. M., et al. The role of integrin receptors in tumor cell adhesion to the microvasculature. In: Orr, F. W.; Buchanan, M. R.; Weiss, L., eds. Microcirculation in cancer metastasis. Boca Raton: CRC Press; 1991:145–167.
Larsen, T.; Sørensen, M. B.; Olsen, R., et al. Effect of scavengers of active oxygen species and pretreatment with acetyl-salicylic acid on the injury to cultured endothelial cells by thrombin-stimulated platelets. In Vitro Cell. Dev. Biol. 25:276–282; 1989.
Marcum, J. M.; McGill, M.; Bastida, E., et al. The interaction of platelets, tumor cells, and vascular subendothelium. J. Lab. Clin. Med. 96:1046–1053; 1980.
Markwardt, F. Hirudin as an inhibitor of thrombin. Methods Enzymol. 69:924–932; 1970.
Martin, W. J. Neutrophils kill pulmonary endothelial cells by a hydrogen-peroxide-dependent pathway: an in vitro model of neutrophil-mediated lung injury. Am. Rev. Respir. Dis. 130:209–213; 1984.
Mehta, P. Potential role of platelets in the pathogenesis of tumor metastasis. Blood 63:55–65; 1984.
Menter, D. G.; Steinert, B. W.; Sloane, B. F., et al. Role of platelet membrane in enhancement of tumor cell adhesion to endothelial cell extracellular matrix. Cancer Res. 47:6751–6762; 1987.
Morrison, A. R.; Winokur, T. S.; Brown, W. A. Inhibition of soybean lipoxygenase by mannitol. Biochem. Biophys. Res. Commun. 108:1757–1762; 1982.
Mustard, J. F.; Kinlough-Rathbone, R. L.; Packham, M. A. Isolation of human platelets from plasma by centrifugation and washing. In: Hawiger, J., ed. Methods in enzymology: platelets, receptors, adhesion, secretion. Part A; New York: Academic Press; 169:3–11; 1989.
Rice, G. E.; Gimborne, M. A., Jr.; Bevilacqua, M. P. Tumor cell-endothelial interactions. Increased adhesion of human melanoma cells to activated vascular endothelium. Am. J. Pathol. 133:204–210; 1988.
Rickles, F. R.; Levine, M.; Edwards, R. L. Hemostatic alterations in cancer patients. Cancer Metastasis Rev. 11:237–248; 1992.
Solberg, S.; Larsen, T.; Jørgensen, L. Differences in reactivity of confluent and nonconfluent cultures of human endothelial cells toward thrombin-stimulated platelets or heparinized salt solution. In Vitro Cell. Dev. Biol. 21:612–616; 1985.
Tang, D. G.; Grossi, I. M.; Chen, Y. Q., et al. 12(S)-HETE promotes tumor-cell adhesion by increasing surface expression of Vβ3 integrins on endothelial cells. Int. J. Cancer 54:102–111; 1993.
Warren, B. A.; Güldner; F.-H. Ultrastructure of the adhesion of HeLa cells to human vein wall. Angiologica 6:32–53; 1969.
Wörner, P. Arachidonic acid-induced chemiluminescence of human platelets: contribution of the prostaglandin and lipoxygenase pathways. Thromb. Haemostasis 46:584–589; 1981.
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Helland, I.B., Klementsen, B. & Jørgensen, L. Addition of both platelets and thrombin in combination accelerates tumor cells to adhere to endothelial cells In vitro . In Vitro Cell.Dev.Biol.-Animal 33, 182–186 (1997). https://doi.org/10.1007/s11626-997-0139-6
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DOI: https://doi.org/10.1007/s11626-997-0139-6