Soluble tissue factor has unique angiogenic activities that selectively promote migration and differentiation but not proliferation of endothelial cells
Section snippets
Materials and methods
Human recombined sTF from E. coli. Coding region for human sTF (TF1-219, residues 1–219) was amplified by PCR using TF cDNA. sTF, constructed and expressed in E. coli BL21/DE3, was purified and refolded thereafter. The clotting activity of refolded sTF was analyzed and verified after reconstitution with phospholipids as described previously [9].
Cell culture. Primary HUVECs were isolated from freshly delivered umbilical cords and grown in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen)
sTF stimulates the migration of endothelial cells
Angiogenesis involves cell migration, proliferation, and tubule formation [6]. The mobilization of endothelial cells at the microvascular level is a prerequisite for the neovascular occurrence [8]. Thus, we first set out to determine the migratory effect of sTF on HMECs and HUVECs using Millicell separate culture plate inserts. After 6 h, the migration of HMECs on collagen-precoated transwell filter in response to the increasing concentrations of sTF was significantly stimulated over that in the
Discussion
TF, a transmembrane receptor protein, is the primary initiator of coagulation and also participates in tumor angiogenesis, growth, and metastasis [2]. Importantly, elevated levels of circulating TF were discovered in the blood of patients with angiogenesis-related tumor, hypercoagulability, and vascular diseases. For example, circulating TF and membrane TF expressed by tumor cells positively correlates with tumorigenesis and angiogenesis in human colorectal carcinoma [4]. Besides, it was
Acknowledgments
This study was supported in part by the General Programs of National Natural Science Foundation of China (Nos. 30670419, 30771083), and the State Key Development Program for Basic Research of China (No. 2006CB910305).
References (22)
- et al.
Oncogenic events regulate tissue factor expression in colorectal cancer cells: implications for tumor progression and angiogenesis
Blood
(2005) - et al.
Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis
Microvasc. Res.
(1977) - et al.
Molecular mechanisms of blood vessel formation
Trends Biochem. Sci.
(1997) - et al.
Nucleolin is a receptor that mediates antiangiogenic and antitumor activity of endostatin
Blood
(2007) - et al.
Deficiency of disulfide bonds facilitating fibrillogenesis of endostatin
J. Biol. Chem.
(2006) - et al.
The angiogenic function of nucleolin is mediated by vascular endothelial growth factor and nonmuscle myosin
Blood
(2006) - et al.
Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis
Cell
(1996) - et al.
Tissue factor-mediated endocytosis, recycling, and degradation of factor VIIa by a clathrin-independent mechanism not requiring the cytoplasmic domain of tissue factor
Blood
(2001) - et al.
Down-regulation of monocyte tissue factor mediated by tissue factor pathway inhibitor and the low density lipoprotein receptor-related protein
J. Biol. Chem.
(1999) - et al.
Extrinsic-pathway activation in cancer with high factor VIIa and tissue factor
Lancet
(1995)
Initiation of coagulation by tissue factor
CRC Crit. Rev. Biochem.
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