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HUVEC monolayers were either left untreated or were treated with 100 ng/ml of VEGF or bFGF, or with 20 ng/ml of PMA every 6 hours for 48 hours.
4H, lane 3 and 4), in contrast, in bFGF stimulated cells (Fig.
The effects of PGs on cell proliferation and collagen production have been widely studied in different cell types [ 13 14 15 16 17 26 ] . TXA 2 has been studied extensively because of its apparent role in atherosclerosis, due to its prothrombotic and mitogenic activities on vascular smooth muscle cells [ 15 16 ] . These mitogenic effects are potentiated by growth factors [ 15 16 27 28 ] . In vascular smooth muscle cells TXA 2 stimulates synthesis of bFGF and increases the expression of the proto-oncogenes c-fos , c-myc , and egr-1 , which are associated with entry into the cell growth cycle [ 15 ] . In addition, TXA 2 increases proliferation of fibroblasts [ 13 ] and smooth muscle-like glomerular mesangial cells [ 14 ] .
Mediators known to activate ECs include inflammatory cytokines, growth factors such as VEGF/VPF and basic fibroblast growth factor (bFGF), extracellular matrix (ECM) proteins such as collagen and fibronectin, and proteases such as MMPs [ 1 2 ] . In vitro and in vivo assays indicate that VEGF signaling promotes increased permeability, cell migration, proliferation, and differentiation, functioning through two EC-specific tyrosine kinase receptor such as Kinase domain receptor (KDR) [ 1 2 3 ] . In the majority of solid tumors, VEGF is upregulated in response to hypoxia, inducing the expression of pro-angiogenic genes that promote tumor angiogenesis, growth and eventual metastasis [ 3 4 5 ] . The expression of both positive and negative factors must be tightly and specifically regulated in a coordinated manner.
A number of in vitro model systems have thus been developed to study EC activation that employs either one or more combinations of ECM molecules including fibrin, fibronectin, collagens, laminins, and Matrigel ®matrices together with PMA and FGF [ 7 8 9 ] . When placed in a 3D type I collagen matrix and stimulated with PMA, VEGF, or bFGF, ECs undergo rapid morphological changes and differentiate into capillary-like tubular networks.
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