Enhanced Cytotoxicity and Mechanistic Responses to Co-Exposure to the Pesticide β-Cypermethrin and the Foodborne Contaminant Ethyl Carbamate in Caco-2 Cells.

چکیده اصلی

Beta-cypermethrin (β-CYP) is a widely used pyrethroid insecticide, whereas ethyl carbamate (EC) is a processing-derived contaminant in fermented foods. This mechanism-oriented in vitro study examined cellular responses to EC, β-CYP, and their combination in Caco-2 cells; the concentrations were substantially higher than typical dietary exposure and were not intended for quantitative risk assessment. Cell viability, apoptosis, cell cycle, wound closure, and mitochondrial membrane potential (ΔΨm) were evaluated via CCK-8, AO/EB staining, flow cytometry, wound-healing, and JC-1 assays. Network toxicology, molecular docking, RNA sequencing (RNA-seq), and quantitative real-time PCR (qRT-PCR) were integrated to generate mechanistic hypotheses. EC and β-CYP each reduced cell viability, wound closure, and ΔΨm in a concentration-dependent manner. To rigorously determine the interaction type, a 2 × 2 factorial design coupled with two-way ANOVA was employed. Co-exposure exhibited an additive trend for cell viability and cell cycle (G0/G1) arrest (interaction p > 0.05), whereas significant synergistic interactions were observed for apoptosis, impaired wound closure, and loss of ΔΨm (interaction p < 0.05). Although CI values for viability were below 1, they were considered only as exploratory references due to extrapolation beyond the observed response range, and were not treated as statistically demonstrated synergism. Bioinformatic analyses implicated stress-response and cell-fate pathways, including PI3K-Akt, MAPK, FoxO, and TGF-β, but do not establish direct target engagement or colorectal cancer causation. Overall, co-exposure enhanced Caco-2 cell injury under the tested conditions. These findings support further mixture-toxicity studies at exposure-relevant concentrations but cannot be directly extrapolated to human dietary risk.

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