Fig. 5

GLPG0187 suppresses breast tumor invasion and metastasis in zebrafish. (A) MDA-MB-231 cells were treated with vehicle, 0.5 ng/ml or 1 ng/ml of GLPG0187 for 48 h with or without 5 ng/ml transforming growth factor (TGF)-β for 2 h and immunoblotted as shown. (B) MDA-MB-231 cells were treated with vehicle, 0.5 ng/ml or 1 ng/ml of GLPG0187 for 48 h with or without 5 ng/ml TGF-β for 6 h and subjected to quantitative PCR analysis. mRNA levels of PAI-1, CTGF, and IL-11 were normalized to GAPDH expression. (C) MDA-MB-231 cells were pretreated for 6 h with 1 ng/ml GLPG0187 or vehicle, then injected into the blood circulation of zebrafish 48-h post fertilization and followed with or without 0.5 ng/ml GLPG0187 treatment, as indicated. Confocal images of overview zebrafish were made 5 days post implantion (dpi). Arrows indicate invasive tumor cells, scale bar: 500 µm. (D) Percentage of embryos displaying invasion and metastasis at 1, 3, and 5 dpi is shown; vehicle versus GLPG0187 P <0.001. (E) Representative images of the zebrafish described in panel D. Experimental micrometastasis was detected in the posterior tail fin at 5 dpi. Arrows indicate invasive tumor cells, scale bar: 100 µm.