Figure 2

Effect of MMP-2 siRNA on ameloblastoma cells. Ameloblastoma cells were transfected with mock, lipofectamine, pRNA-neg, pRNA-MMP-2 (1 μg), pRNA-MMP-2 (2 μg), and pRNA-MMP-2 (3 μg) for the indicated times. GAPDH was used as an internal control to normalize the amounts of protein or mRNA. The relative protein or mRNA level of MMP-2 was depicted as the ratio of the density of MMP-2 to GAPDH for the same time point. The results shown are representative of triplicate experiments. *Compared with the mock, lipifectamine or vector controls, respectively, p < 0.05; #Compared with the 1 μg plasmid transfection group, p < 0.05. (A) Western blots were used to analyze MMP-2 protein levels in MMP-2 siRNA-transfected ameloblastoma cells. MMP-2 siRNA transfection decreased MMP-2 protein expression levels in a dose- and time-dependent manner compared with mock and scrambled vector controls. (B) RT-PCR was employed to detect the transcription of MMP-2 after MMP-2 siRNA transfection. Transcript levels of MMP-2 in MMP-2 siRNA-transfected cells were significantly reduced in a dose- and time-dependent manner compared with cells transfected with mock and lipofectamine and pRNA-neg. (C) Zymographic analysis for MMP-2 activity in the conditioned medium of ameloblastoma cells transfected with pRNA-MMP-2 for 96 h. Both latent and active form of MMP-2 and MMP-9 were detected in the gelatin zymogram. MMP-2 siRNA transfection inhibited MMP-2 activity in a dose-dependent manner in ameloblastoma cells.