Figure 5

(A) Representative western blot analysis of COX-2 expression by VEGF-treated B16M cells. Cultured B16M cells were given 10 or 100 ng/ml murine recombinant VEGF for 4 h. Cell lysates were collected and assayed for COX-2 and β-tubulin levels by western immunoblot. (B) Effect of celecoxib on the proadhesive response of VEGF-treated B16M cells on immobilized VCAM-1. B16M cells received 1 μg/ml celecoxib for 30 min and then incubated with 100 ng/ml rmVEGF for 4 h. In other experiments B16M cells were given 10 or 100 ng/ml of PGE2 for 2 h. Then, cell adhesion assay to rhVCAM-1-coated plate was performed. Data are expressed as mean percent of added labeled-cells binding to quadruplicate wells ± SD. Statistical significance by ANOVA and Bonferroni's post-hoc test: *P < 0.01 as compared with basal medium-treated B16M cells; **P < .001 as compared with VEGF-treated B16M cells. C) Effect of celecoxib and anti-VEGF on the proadhesive response of A375M cells to bone marrow-conditioned media on immobilized VCAM-1. Human A375M cells received 1 μg/ml celecoxib for 30 min and then incubated in the presence of basal medium, hBMSC-CM, LPS-treated hBMSC-CM or rhVEGF (10 ng/ml) for 4 h. Then, cell adhesion assay to a rhVCAM-1-coated plate was performed. Data are expressed as mean percent of added labeled-cells binding to quadruplicate wells ± SD. Statistical significance by ANOVA and Bonferroni's post-hoc test: *P < 0.01 as compared with basal medium-treated A375M cells; **P < 0.01 as compared with BMSC-CM-; +P < 0.01 as compared with LPS-treated BMSC-CM-treated A375M cells; #P < 0.01 as compared with rhVEGF-treated A375M cells.