Paeonol Oxime Inhibits bFGF-Induced Angiogenesis and Reduces VEGF Levels in Fibrosarcoma Cells
Figure 5
PO downregulates the expression of VEGF and the PI3K/AKT signaling in HT-1080 cells.
(A) HT-1080 cells were treated with PO for 24 h and the cell viability was assessed by MTT assay. (B) The conditioned medium from HT-1080 cells treated with or without PO for 24 h was used for VEGF measurement using an ELISA kit. Level of VEGF was normalized to the cell number of each sample after 24 h of PO treatment. ***, p<0.001 versus control. (C) HT-1080 cells were treated with PO (0, 6.25 or 12.5 µg/ml) for 24 h. The levels of VEGF, PI3K, phospho-AKT and AKT were determined by Western blotting. (D) HT-1080 cells were treated with LY294002 or PO for 24 h and subjected to Western blotting for VEGF, phospho-AKT and AKT. (E) Cells were treated with LY294002 or PO as described in (D) and incubated with anti-phospho-AKT and FITC conjugated secondary antibodies. The immunostained cells were mounted in medium containing DAPI and visualized under a Carl Zeiss LSM5 confocal microscope.