Figure 4.

Up‐regulating ADAM8 induced EMT effect and enhanced the invasive ability of colon cancer cells. A, The level of ADAM8 at mRNA and protein in six colon cancer cell lines (SW480, SW620, DLD1, HCT116, HCT8 and HT29) was analysed by qRT‐PCR and Western blotting. B, HCT116 was transfected pcDNA3.1, 3 μg or 5 μg Flag‐ADAM8 for 48 h and cells were harvested for Western blotting and qRT‐PCR analysis. C, D and E, HCT116 cells treated with 5 μg Flag‐ADAM8 and 5 ng/mL TGF‐β for 48 h. Western blotting and qRT‐PCR were employed to assess the level of ADAM8, E‐cadherin, N‐cadherin, and Vimentin at protein and mRNA. F and G, Transwell invasion assay was used to assess the invasiveness of HCT116 treated with Flag‐ADAM8 and TGF‐β for 48 h. The average number of cells invaded through the filter was counted. The data were obtained from three independent experiments. The data were presented as the mean ± standard deviation and two‐way ANOVA. The expression of all proteins was quantified by Image J software and normalized to the quantified value of GAPDH. *P < .05; **P < .01; ***P < .001; ns, no significant