Figure 2

Cell-based evidence that MT1-MMP directly regulates COX-2 expression in U87 glioma cell lines. Monolayers or neurospheres from glioblastoma-derived cells were either Mock-transfected, transfected with a cDNA plasmid encoding MT1-MMP (Wt), or transfected with an siRNA (Si) against MT1-MMP as described in the Methods section. (A) Gelatin zymography was performed to monitor the extent of latent proMMP-2 and active MMP-2 expression from the conditioned media of the serum-starved cells. Cell lysates were isolated from U87 glioblastoma-derived cells and SDS-PAGE performed (20 μg protein/well), followed by Western-blotting and COX-2 or GAPDH immunodetection. (B) Total RNA was isolated from monolayers (white bars) or neurospheres (black bars) of U87 Mock-transfected cells, or from U87 cells transfected with MT1-MMP cDNA or siRNA against MT1-MMP, and reverse-transcribed as described in the Methods section. Quantitative PCR was performed in order to monitor COX-2 gene expression levels.