Fig.3. CFI-402257 suppresses growth of MM cells in vitro and has no effect on normal mesothelial cells.

Five human MM cell lines (a) and three primary cultures of mesothelial cells derived from non-cancer patients (b) were plated 3 × 10³ cells/well of 96-well plate and treated with increasing concentrations of CFI-402257. Alamar Blue (Thermo Fisher, MA, US) viability assay was performed after 5 days of treatment. EC50 values were calculated using GraphPad PRISM software. (c) The ability of MM cell lines to form colony in soft agar was evaluated under treatment with 10 nM and 100 nM CFI-402257 or DMSO. Total of 20 (Phi) and 10 (Mill) colonies were measured from 3 wells per condition. The graph represents average colony size expressed as the percentage of vehicle. Pictures are representative of the experiment performed with Phi cells. (d) Mill and Hmeso cell lines were plated 3 × 10³ cells/well of 96-well plate and treated with 10µM Cisplatin and/or 10nM CFI-402257. After 72 hr, Alamar Blue viability assay was performed. To determine the effects of drug combinations CDI was calculated⁵⁸ as follows: CDI = AB/(A × B), where AB is the ratio of the measured effect of combination to control group; A or B is the ratio of the single agent to control group. Thus, CDI <1, = 1 or >1 indicates that the drugs are synergistic, additive or antagonistic, respectively. CDI <0.7 indicates that the drug is significantly synergistic.