Figure 1

COX-2-directed migration of human chondrosarcoma cells. JJ012 cells were transfected with IPTG/COX-2 expression plasmid or control vector for 24 hr followed by stimulation with IPTG (5 mM) for 24 hr, the COX-2 expression, PGE₂ production and migration activity were determined by Western blot analysis (A), ELISA assay (B) and Transwell (C). JJ012 cells were transfected with IPTG/COX-2 expression plasmid or control vector for 24 hr, and pretreated with valeryl salicylate (20 μM), celebrex (10 μM) or NS-398 (20 μM) for 30 min followed by stimulation with IPTG (5 mM), and in vitro migration was measured with the Transwell after 24 hr (D). JJ012 cells were incubated with various concentrations of PGE₂, and in vitro migration activity measured with the Transwell after 24 hr (E). Total RNA were extracted from normal cartilage (lines 1-4), chondrosarcoma patients (lines 5-8) or from chondrosarcoma cell lines (SW1353 and JJ012), and subjected to qPCR analysis for COX-2 (F). The migration activity of each cells measured in vitro with the Transwell chamber after 24 h showed a significantly higher migration activity in primary chondrosarcoma and chondrosarcoma cell lines as compared with primary chondrocyte (G). Results are expressed as the mean ± S.E. *, p < 0.05 compared with control; #, p < 0.05 compared with PGE₂-treated group.