Soluble vascular endothelial growth factor (VEGF) receptor-1 inhibits migration of human monocytic THP-1 cells in response to VEGF
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We aimed to investigate the regulation and contribution of vascular endothelial growth factor (VEGF) and sFlt-1(1–3) to human monocytic THP-1 migration.
Materials and methods
Ad-sFlt-1/FLAG, a recombinant adenovirus carrying the human sFlt-1(1–3) (the first three extracellular domains of FLT-1, the hVEGF receptor-1) gene, was constructed. L929 cells were infected with Ad-sFlt-1/FLAG and the expression of sFlt-1 was detected by immunofluorescent assay and ELISA. Corning® Transwell® Filter Inserts containing polyethylene terephthalate (PET) membranes with pore sizes of 3 μm were used as an experimental model to simulate THP-1 migration. Five VEGF concentrations (0, 0.1, 1, 10 and 100 ng/ml), four concentrations of sFlt-1(1–3)/FLAG expression supernatants (0.1, 1, 10 and 100 ng/ml), and monocyte chemoattractant protein-1 (MCP-1, 10 ng/ml) were used to test the ability of THP-1 cells to migrate through PET membranes.
The sFlt-1(1–3) gene was successfully recombined into Ad-sFlt-1/FLAG. sFlt-1(1–3) was expressed in L929 cells transfected with Ad-sFlt-1/FLAG. THP-1 cell migration increased with increasing concentrations of VEGF, while cell migration decreased with increasing concentrations of sFlt1(1–3)/FLAG. sFlt1(1–3)/FLAG had no effect on MCP-1-induced cell migration.
This study demonstrated that VEGF is able to elicit a migratory response in THP-1 cells, and that sFlt-1(1–3) is an effective inhibitor of THP-1 migration towards VEGF.
KeywordsVEGF sFlt-1(1–3) gene Human monocytic THP-1 Migration
- 13.Kim I, Moon SO, Kim SH, Kim HJ, Koh YS, Koh GY. Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells. J Biol Chem. 2001;276:7614–20.PubMedCrossRefGoogle Scholar
- 15.Kondo K, Hiratsuka S, Subbalakshmi E, Matsushime H, Shibuya M. Genomic organization of the flt-1 gene encoding for vascular endothelial growth factor (VEGF) receptor-1 suggests an intimate evolutionary relationship between the 7-Ig and the 5-Ig tyrosine kinase receptors. Gene. 1998;208:297–305.PubMedCrossRefGoogle Scholar