Interaction effects of enniatin B, deoxinivalenol and alternariol in Caco-2 cells

Abstract Enniatin B (ENN B), deoxinivalenol (DON) and alternariol (AOH) are secondary metabolites of filamentous fungi. These mycotoxins are contaminants of vegetables and cereals. They are cytotoxic and their effects are enhanced by their mixtures. The objectives of this study were to compare the cytotoxicity of ENN B, DON and AOH alone or in combination in human adenocarcinoma (Caco-2) cells and to evaluate the type of interactions of mycotoxin mixtures by the isobologram analysis. Cells were treated with concentrations ranging from 1.85 to 90 μM (AOH) and from 0.312 to 10 μM (for ENN B and DON), individually and in combination of two and three mycotoxins (from 1.85 to 30 μM for AOH and from 0.312 to 5 μM for ENN B and DON). The relation ratios between the mixtures DON + ENN B was 1:1; AOH + DON and ENN B + AOH was 1:6, and for the tertiary combination DON, ENN B and AOH 1:1:6. The IC 50 value of ENN B and DON were 3.87 and 5.54 μM, respectively. No IC 50 values were obtained for the AOH at any time tested in Caco-2 cells. With the isobologram the type of interaction between mycotoxin was evaluated. Synergistic, antagonistic and addictive effect was observed for the combination studied depending on the concentration affected. Mycotoxins combinations reduce cellular viability in the following increasing order: (DON + ENN B) > (ENN B + AOH) > (DON + AOH) > (DON + AOH + ENN B)