Figure 2

CCL3-directed migration activity of human chondrosarcoma cells involves up-regulation of MMP-2. (A-B): Cells were incubated with CCL3 for 24 h, and in vitro migration was measured either by Transwell (A) or by a wound-healing assay (B). (C): JJ012 cells were incubated with CCL3 for 24 h, and MMP-1, MMP-2, MMP-3, MMP-9, and MMP-13 mRNA levels were determined using qPCR. (D): JJ012 cells were incubated with CCL3 for the indicated periods or with the indicated doses of CCL3, and cell lysates and supernatants were collected. MMP-2 protein levels in cell lysates and enzymatic activity in supernatants were determined by western blotting and zymography. (E-F): JJ012 cells were transfected with MMP-2 siRNA or were pre-treated with MMP-2 inhibitor, and in vitro migration was measured with the Transwell (E) or the wound-healing assay (F). (G-H): In vitro migration activity of JJ012/control-shRNA and JJ012/CCL3-shRNA cells was measured with Transwell and wound-healing assays. (I): The protein levels of CCL3 and MMP-2 of in JJ012 cells transfected with control-shRNA or CCL3-shRNA was measured by western blotting. The results are expressed as the mean ± SE. *P < 0.05 compared with control. ^#P < 0.05 compared with CCL3-treated group.