FIG. 11.

Both AOAA and vascular endothelial growth factor (VEGF) neutralization inhibit tumor-induced endothelial cell migration. Tumor-induced endothelial cell migration was tested in vitro in a co-culture assay involving human umbilical endothelial cells (EAhy926) and human colon cancer cells (HCT116). HCT116 cells were seeded to confluence in the lower chamber of a 6.5 mm Transwell insert (8.0 μm pore); while EAhy926 were serum starved for 5 h, detached by Trypsin-EDTA, re-suspended in serum-free media, and added to the upper chamber (10⁵ cells/well). Cells were allowed to migrate for 4 h. Migrated cells were fixed with Carson's fixative, stained by 0.33% Toluidine blue, and quantified by visual counting. AOAA (1 mM) and antihuman VEGF polyclonal antibody (5 μg/ml) were added to the HCT116 culture at 45 min (37°C) before the addition of the endothelial cells to the upper chamber (*p<0.05 vs. vehicle and ^#p<0.05 vs. HCT116). The comparable degree of inhibition of endothelial cell migration by AOAA and of VEGF neutralization should be noted. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars