Article Figures & Data
Figures
- Figure 1.
A, structure of FC. B, cytotoxic effect of FC on HUVECs under nonproliferative conditions. HUVEC cells were exposed to FC in M199 medium containing 5% FBS (without heparin and growth factors) for 24 h and the proportion of metabolically viable cells was assessed by the XTT assay. Mean ± SEM, n = 3. ***, P < 0.01; ***, P < 0.001 compared with control. C, effect of FC on VEGF-induced tube formation (18-h stimulation in the absence or presence of FC). Data in graph were presented as mean ± SEM, n = 5 fields. ###, P < 0.001 compared with basal control; ***, P < 0.001 compared with VEGF-stimulated group. D, effect of FC on VEGF-A (10 ng/mL)–induced proliferation of HUVECs for 48 h. The relative number of metabolically viable cells were estimated by the XTT assay. Data were means ± SEM, n = 3 independent wells. ###, P < 0.001 compared with basal control; *, P < 0.05 compared with VEGF-stimulated group.
- Figure 2.
A, effect of FC on VEGF-induced cell motility (wound-healing test). Confluent HUVEC monolayers on 0.1% gelatin-coated six-well plates were scratch wounded. The cells were treated with various concentrations of FC in M199 with 5% FBS, 10 ng/mL VEGF, and 5 units/mL heparin for 16 h. Representative fields were photographed, ×100 magnification. Graph shows the quantitative effect of FC on VEGF-induced HUVEC motility. Data were presented as mean ± SEM, n = 3 wells. ###, P < 0.001 compared with basal control; **, P < 0.01; ***, P < 0.001 versus VEGF-stimulated group. B, effect of FC on VEGF-induced invasion of HUVEC through Matrigel in 12 h. Data were presented as means ± SEM n = 3 wells. ###, P < 0.001 compared with basal control; **, P < 0.01 compared with VEGF-stimulated group. C, effect of FC on VEGF-induced MMP-2 secretion from HUVECs after 20 h examined by zymography. D, effect of FC on VEGF-induced vessel sprouting ex vivo from rat aortic segments. Rat aortic segments were cultured on Matrigel and treated with VEGF (20 ng/mL) in the absence or presence of FC for 6 d. Graph shows the endothelial sprouts counts as the relative ratio to VEGF-stimulated control as 100. Data were presented as mean ± SEM, n = 3 aortas. ##, P < 0.01 compared with basal control; *, P < 0.05 and **, P < 0.01 compared with VEGF-stimulated control.
- Figure 3.
Effect of FC on the in vivo growth of LLC allograft tumors in mice and on angiogenesis and proliferation indices. LLC cells (5 × 105) were injected s.c. into the right flank of C57BL/6 mice. After 5 d, mice were given i.p. injection of FC (0.1 mg/kg and 1 mg/kg) or PBS (control) once a day. A, body weights of mice. B, tumor growth over time. Mean ± SD, n = 10 mice per group. *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared with control. C, CD34 index (angiogenesis). D, Ki67 index (proliferation), E, TUNEL index (apoptosis). *, P < 0.05; **, P < 0.01 compared with control. Mean ± SD, n = 10.
- Figure 4.
A, effect of FC on the binding of VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1) to immobilized VEGF. Data were presented as means ± SEM, n = 3. **, P < 0.01; ***, P < 0.001 compared with control. B, Western blot analyses of effect of FC on VEGFR-signaling kinases. HUVECs were pretreated with FC for 30 min and further stimulated with 10 ng VEGF/mL for 10 min. C, in vitro Src kinase activity of FC-treated HUVEC cells prepared as in B.
- Figure 5.
The potential binding modes (poses) of FC at the active site of Src tyrosine kinase (A–C) and FAK (D–F) by docking simulation. A and D, hydrophobic pocket near the ATP-binding site in Src and FAK, respectively. Connolly surface was generated for the protein by MOLCAD and colored by cavity depth, in which orange means deep cavity of the hydrophobic pocket. B and C, two possible binding modes of FC in Src. E and F, two possible binding modes of FC in FAK. FC is displayed in ball-and-stick and its carbon colors are magenta/purple in Src and orange/white in FAK, respectively. The Connolly surface was Z-clipped and the nonpolar hydrogen atoms are not displayed for clarity. Hydrogen bonds are displayed as yellow dashed lines and the participating residues are marked.
Additional Files
Supplementary Data, Lee et al.
Files in this Data Supplement:
Volume 9, Issue 2
