Figure 5.

MIM-4 could act as an adjuvant when associated with etoposide or resveratrol, in an in vitro 3D-spheroid model of CRC. (A) Spheroids were cultivated in standard conditions and treated for 24, 48 or 72 h with either ETP (orange curve) or RSV (pink curve), at 50 µM each. Spheroids cultivated in standard conditions without any treatment were used as a positive control (Ctrl). Spheroid volumes were calculated and the growth evolution was represented as the fold change of the initial spheroid volume at the time of the treatment initiation. Results are presented as the growth mean ± SD for each condition (n = 3). (B) The cytotoxicity of both ETP and RSV was appraised in the same conditions as described in (A). The results are presented as the mean normalized green fluorescence intensity (NGFI) ± SD from CellTox green fluorescent dye, for each condition (n = 3). NGFI was calculated as described in Section 4.4.3. Spheroids were cultivated in the presence of RSV (C) or ETP (D) for 24, 48, or 72 h, concomitantly with either the Veh. (grey histograms) or MIM-4 (squared histograms) and the spheroid volumes were calculated in each condition. The results are presented as the mean ± SD of the volume percentage, the Veh. conditions being set at 100% (n = 3). The cytotoxicity of the association of RSV with either Veh. or MIM-4 (E) and the cytotoxicity of the association of ETP with either Veh. or MIM-4 (F) was appraised in the same assessed conditions as in (B,C). The results are presented as the mean ± SD of the NGFI percentage, the Veh.-conditions being set at 100% (n = 3).