Figures & data
Table 1. CA I, II, IX and XII inhibition data with sulfamates FC8–399A and S4 in comparison with AAZ as standard by a stopped-flow CO2 hydrase assayCitation46.
Figure 1. CA IX expression and cytotoxic effect of FC9–399A and S4 inhibitors on Me30966 cells. (a) Western blot analysis of CA IX expression in total cellular extract of Me30966 cultured in unbuffered, line 1 and buffered, line 2 conditions. As a control for protein loading the membranes were blotted with anti-GAPDH antibody (b). Me30966 cells were incubated in buffered and unbuffered medium, treated for 24 h with 5, 10, 50 and 100 μM of FC9–399A (b) and S4 (c). Columns, mean percentages of cell death of three independent experiments run in triplicate; bars indicate SD. (*) indicates p < 0.001 (compared to single treatments).
Figure 2. Combined effect of Lansoprazole and FC9–399A or S4 treatment on Me30966 cell growth. Me30966 cells cultured in unbuffered conditions, were treated with LAN for 24 or 48 h. Then, the cells were treated for additional 24 h with FC9–399A or S4 at two different concentrations: 10 and 50 μM. Cell proliferation was evaluated by the 405 nm absorbance measured by a spectrophotometer EL × 800. Columns, mean percentages of cell growth of three independent experiments run in triplicate; bars indicate SD. (*) indicates p < 0.001 (compared to single treatments).
Figure 3. Combined effect of Lansoprazole and FC9–399A or S4 treatment on cell death and CA IX protein expression in Me30966 cells. (a) Me30966 cells cultured in unbuffered conditions, were treated with LAN for 24 h. Then, the cells were treated for additional 24 h with FC9–399A or S4 at two different concentrations: 10 and 50 μM. Tumor cell death was evaluated by Trypan blue exclusion method. Columns, mean percentages of cell death of three independent experiments run in triplicate; bars indicate SD. (*) indicates p < 0.001 (compared to single treatments). (b) Western blot analysis of CA IX expression in total cellular extract of (1) Me30966; (2) Me30966 treated with LAN for 24 h; (3) Me30966 treated with FC9–399A for 24 h; (4) Me30966 treated with S4 for 24 h; (5) Me30966 pretreated with LAN and then treated with FC9–399A (50 μM) for additional 24 h; (6) Me30966 pretreated with LAN and then treated with S4 (50 μM) for additional 24 h. As a control for protein loading, the membranes were blotted with anti-tubulin antibody.
Figure 4. Chemical structure of ureido-sulfamate compounds FC9–399A and S4 and Lansoprazole.
