Figure 7.

MMP9 overexpression increased invasion of SUM225 cells and made the cells more sensitive to everolimus inhibition of invasion. A, MMP9 and p-P70S6K1 levels in control vector or MMP9 expression vector transfected SUM225 cells, which were treated wtih 10 nM everolimus for 24 hours. GAPDH is loading control. B, Everolimus inhibited invasion of 3D-spheroids of SUM225 control and MMP9 overexpressing cells through Matrigel. The representative images were acquired with a 20× objective. Scale bars: 50 μm. C, The invasive percentage was calculated by No. of invasive 3D-organoid / No. of total 3D-organoid X 100. Data are presented as means ± SEM, n=3, *p<0.05 and ## p<0.01 represents the comparison between SUM225 cells transfected with empty vector and everolimus treatment or MMP9 overexpression, or both by one-way ANOVA.