Combined exposure to T-2 and HT-2 mycotoxins induces ferroptosis through down-regulation of SLC7A11/GPX4 and NAD(P)H/FSP1/CoQ10 pathways in Caco-2 cells.

The co-contamination of T-2 and HT-2 mycotoxins in food poses significant health risks to humans. Our previous studies demonstrated that combined exposure to T-2 and HT-2 induces inflammatory responses and damage of intestinal barrier functions. However, the involvement of ferroptosis remains poorly understood together with other potential pathways. In this study, we investigated the combined toxic effects of T-2 and HT-2 and explored the mechanisms of action focusing on excessive reactive oxygen species (ROS)-induced ferroptosis in colorectal cancer (Caco-2) cells. In general, significantly elevated levels of ROS, lipid reactive oxygen species (L-ROS), iron content, and mitochondrial structural damage, and altered expression of six ferroptosis-related marker genes and proteins were observed in cells treated with T-2 and HT-2 individually and in combination. Furthermore, supplementation with the ferroptosis inhibitor Ferrostatin-1 (Fer-1) and ferroptosis suppressor protein 1 inhibition (iFSP1) effectively modulated oxidative stress imbalance, mitochondrial injury, and ferroptotic responses. These findings indicate that combined exposure to T-2 and HT-2 induces ferroptosis, primarily through the Cystine/GPX4 and NAD(P)H/FSP1/CoQ10 regulatory pathways. This study provides a theoretical basis for understanding mycotoxin-induced ferroptosis and offers valuable insights into the potential health risks associated with the combined toxicity of T-2 and HT-2.