Figure 6.

P38-MAPK activation regulated invasion, EMT and the expression of Bcl-2, c-Myc, MMP9 and cytokines in DU145R80. (a) Invasion capability of DU145R80 cells untreated or treated with SB (30 μℳ) for 24 h was evaluated as the ability to invade matrigel coated chambers (see Materials and Methods). Values are means±S.D. from three independent experiments performed in duplicates and statistical analysis is reported (*P<0.001). (b) MMP-9 concentration was evaluated in the culture media of DU145R80 cells untreated or treated with SB (30 μℳ) for the indicated time points after seeding, by a multiplex ELISA-based immunoassay assay, and were plotted with box-and-whisker graphs. The boxes extend from the 25th to the 75th percentile and the line in the middle is the median. The error bars extend down to the lowest value and up to the highest. (c) IL-12, VEGF, Eotaxin-1, Bcl-2, MMP9 and ZEB1 mRNA expression was evaluated by RT-PCR in DU145R80 cells untreated or treated with SB (30 μℳ) for 6 h. Values are means±S.D. from three independent experiments performed in duplicates and statistical analysis is reported (*P<0.01). (d) Western blot analysis of Bcl2, c-Myc, Keratin 8/18, E-cadherin and ZEB1 proteins expression evaluated in DU145R80 cells untreated or treated with SB (30 μℳ) for 72 h of cell culture. GAPDH and γ-tubulin were used as protein loading control