Abstract
The growth of primary tumours beyond a critical mass is dependent on angiogenesis. The switch to the angiogenic phenotype involves changes in the local equilibrium of cytokines with either pro- or anti-angiogenic properties. Vascular Endothelial Growth Factor (VEGF) is one of the major positive regulators of tumour angiogenesis. Serum VEGF is, in cancer patients, correlated with worse prognosis. Recent evidence suggests that platelets are the main contributors of serum VEGF. We demonstrate, ultrastructurally and with immunofluorescence techniques, the alpha granule and membranous localisation of VEGF and provide further evidence for the role of platelets, both in healthy individuals as in patients with locally and advanced breast cancer, in the storage of circulating VEGF. We also demonstrate that, linear with tumoural progression, platelets accumulate more VEGF. Enhanced production in bone marrow platelet progenitors as well as endocytosis of circulating VEGF by platelets and/or megakaryocytes could explain the higher VEGF load in platelets from advanced cancer patients. This study provides further evidence for a role of platelets in transporting VEGF.
Similar content being viewed by others
References
Conn G, Bayne ML, Sodermann DD et al. Amino acid and cDNA sequences of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor. Proc Natl Acad Sci USA 1990; 87(7): 2628-32.
Tischer E, Mitchell R, Hartman T et al. The human gene for vascular endothelial growth factor: Multiple protein forms are encoded through alternative exon splicing. J Biol Chem 1991; 266: 11947-54.
Veikkola T, Karkainen M, Claesson-Welsh L, Alitalo K. Regulation of angiogenesis via vascular endothelial growth factor receptors. Cancer Res 2000; 60: 203-12.
Benjamin L, Golijanin D, Itin A et al. Selective ablation of immature blood vessels in established human tumors follows vascular endothelial growth factor withdrawal. J Clin Invest 1999; 103: 159-65.
Yamagishi S, Yonekura H, Yamamoto Y et al. Vascular endothelial growth factor acts as a pericyte mitogen under hypoxic conditions. Lab Invest 1999; 79(4): 501-9.
Clauss M, Weich H, Breier G et al. The vascular endothelial growth receptor Flt-1 mediates biological activities. Implications for a functional role of placental growth factor in monocyte activation and chemotaxis. J Biol Chem 1996; 271: 17629-34.
Gruber BL, Marchese MJ, Kew R. Angiogenic factors stimulate mast-cell migration. Blood 1995; 86(7): 2488-93.
Frank S, Hubner G, Breier G et al. Regulation of VEGF expression in cultured keratinocytes. Implications for normal and impaired wound healing. J Biol Chem 1995; 270: 12607-13.
Cohen T, Nahari D, Weiss C et al. Interleukin-6 induces the expression of vascular endothelial growth factor. J Biol Chem 1996; 271: 736-41.
Dvorak HF, Brown LF, Detmar M, Dvorak AM. Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol 1995; 146(5): 1029-39.
Olofsson B, Pajusola K, Kaipanen A et al. Vascular endothelial growth factor B, a novel growth factor for endothelial cells. Proc Natl Acad Sci USA 1996; 93: 2576-81.
Joukov V, Pajusola K, Kaipanen A et al. A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt-4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases. EMBO J 1996; 15: 290-8.
Yamada Y, Nezu J, Shimane M. Molecular cloning of a vascular endothelial growth factor, VEGF-D. Genomics 1997; 42: 483-8.
Carmeliet P, Ferreira V, Breier G et al. Abnormal blood vessel development and lethality in mice lacking a single VEGF allele. Nature 1996; 380: 435-9.
Ferrara N. Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney Int 1999; 56: 794-814.
Dirix LY, Vermeulen PB, Pawinsky A et al. Elevated levels of the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in sera of cancer patients. Br J Cancer 1997; 76: 238-43.
Leung DW, Cachianes G, Kuang W-J et al. Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 1989; 246: 1306-09.
Warren RS, Yuan H, Matli MR et al. Induction of vascular endothelial growth factor by insulin-like growth factor 1 in colorectal carcinoma. J Biol Chem 1996; 271: 29483-88.
Salgado R, Vermeulen PB, Benoy I et al. Platelet number and interleukin-6 correlate with VEGF but not with bFGF serum levels of advanced cancer patients. Br J Cancer 1999; 80(5/6): 892-7.
Vermeulen PB, Salven P, Benoy I Blood platelets and serum VEGF in cancer patients. Br J Cancer 1999; 79(2): 370-3.
Banks RE, Forbes MA, Kinsey SE et al. Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets: Significance for VEGF measurements and cancer biology. Br J Cancer 1998; 77(6): 956-64.
Möhle R, Green D, Moore M Constitutive production and thrombin-induced release of vascular endothelial growth factor by human megakaryocytes and platelets. Proc Natl Acad Sci USA 1997; 94: 663-8.
Katoh O, Tauchi H, Kawaishi K Expression of the Vascular Endothelial Growth Factor (VEGF) receptor gene, KDR, in hematopoietic cells and inhibitory effect of VEGF on apoptotic cell death caused by ionizing radiation. Cancer Res 1995; 55: 5687-92.
Wartiovaara U, Salven P, Mikkola H et al. Peripheral blood platelets express VEGF-C and VEGF which are released during platelet activation. Thromb Haemost 1998; 80(1): 171-5.
Salven P, Orpana A, Joensuu H. Leucocytes and platelets of patients with cancer contain high levels of vascular endothelial growth factor. Clin Cancer Res 1999; 5: 487-91.
Bogers JJ, Nibbeling HA, Deelder HM, Van Marck E. Quantitative and morphological aspects of Unicryl versus Lowicryl K4M embedding in immunoelectron microscopic studies. J Histochem Cytochem 1996; 44(1): 43-48.
White JG. The ultrastructural and regulatory mechanisms of blood platelets. Blood Platelet, Function and Medicinal Chemistry. Amsterdam: Elsevier Science Publishers 1984; 15-59.
Harrison P, Cramer M. Platelet α-granules. Blood reviews 1993; 7: 52-62.
Benoy I, Vermeulen P, Wuyts H, Dirix L. Vascular Endothelial Growth Factor (VEGF) serum concentrations change according to the phase of the menstrual cycle. Eur J Cancer 1998; 34(8): 1298-9.
Gunsilius E, Petzer A, Stockhammer G et al. Thrombocytes are the major source for soluble vascular endothelial growth factor in peripheral blood. Oncology 2000; 58: 169-74.
Verheul HMW, Hoekman K, Luykx-de Bakker S et al. Platelet: Transporter of vascular endothelial growth factor. Clin Cancer Res 1997; 3: 2187-90.
Penington DG, Streatfield K. Heterogeneity of megakaryocytes and platelets. Ser Haematol 1975; 8(1): 22-48.
Harrison P, Savidge GF, Cramer EM. The origin and physiological relevance of alfa-granules adhesive proteins. Br J Haem 1990; 74: 125-30.
Elscolar G, White JG. The platelet open canalicular system: A final common pathway. Blood Cells 1991; 17: 467.
Frojmovic MM, Milton JG. Physical, chemical and functional changes following platelet activation in normal and giant platelets. Blood Cells 1983; 9: 359.
Cramer E, Berger G. Immunoelectron microscopic changes of the platelet plasma membrane after activation. Blood Coagul Fibrinolysis 1996; 7: 172-77.
Handagama P, Shuman M, Bainton D. Incorporation of intravenously injected albumin, immunoglobulin G and fibrinogen in guinea pig megakaryocytes granules. J Clin Invest 1989; 84: 73-82.
Harrison P, Wilbourn B, Debili N et al. Uptake of plasma fibrinogen into the alpha granules of human megakaryocytes and platelets. J Clin Invest 1989; 84: 1320-4.
Zucker-Franklin D. Endocytosis by human platelets, metabolic and freeze-fracture studies. J Cell Biol 1981; 91: 706-15.
George J, Saucerman S. Platelet IgG, IgM, and Albumin: Correlation of platelet and plasma concentrations in normal subjects and in patients with ITP or dysproteinemia. Blood 1988; 72(1): 362-5.
Estrov Z, Talpaz M, Mavligit G et al. Elevated plasma thrombopoietic activity in patients with metastatic cancer-related thrombocytosis. Am J Med 1995; 98: 551-8.
Ray MR, Ghosh A, Nandi SK. Stimulation of thrombopoiesis in mice bearing experimental tumors. Neoplasms 1991; 38(1): 69-76.
Imai T, Koike K, Kubo T. Interleukin-6 supports human megakaryocytic proliferation and differentiation in vitro. Blood 1991; 78: 1969-74.
Bobik R, Hong Y, Breier G et al. Thrombopoietin stimulates VEGF release from c-MPL expressing cell lines and haematopoietic progenitors. FEBS Lett 1998; 423(1): 10-14.
Kema I, de Vries E, Schellings A et al. Improved diagnosis of carcinoid tumors by measurement of platelet serotonin. Clin Chem 1992; 38(4): 534-40.
Page CP. The involvement of platelets in non-thrombotic processes. Trends Pharmacol Sci 1998; 9: 66-71.
Monreal M, Fernandez-Llamazares J, Piñol M et al. Platelet count and survival in patients with colorectal cancer: A preliminary study. Thromb Haemost 1998; 79(5): 916-18.
Gücer F, Moser F, Tamussino K et al. Thrombocytosis as a prognostic factor in endometrial carcinoma. Gynecol Oncol 1998; 70: 210-14.
Pearlstein E, Abrogio C, Karpatkin S. Lack of effect of PGI2 on the development of pulmonary metastases in mice following the intravenous injection of three syngeneic tumours: Inhibition of metastases with anti-platelet antibody. Proc Am Fed Clin Res, Washington DC, 1984; 32(2): 421A.
Pinedo HM, Verheul HMW, D'Amato RJ, Folkman J. Involvement of platelets in tumour angiogenesis? Lancet 1998; 352: 1775-77.
Salgado R, Benoy I, Weytjens R et al. Serum vascular endothelial growth factor load and interleukin-6 in cancer patients. Br J Cancer 2000; 82(11): 1896.
O'Byrne KJ, Dobbs N, Harris AL. Serum vascular endothelial growth factor load and interleukin-6 in cancer patients. Br J Cancer 2000; 82(11): 1895.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Salgado, R., Benoy, I., Bogers, J. et al. Platelets and vascular endothelial growth factor (VEGF): A morphological and functional study. Angiogenesis 4, 37–43 (2001). https://doi.org/10.1023/A:1016611230747
Issue Date:
DOI: https://doi.org/10.1023/A:1016611230747