Monohaloacetonitriles induce cytotoxicity and exhibit different mode of action in endocrine disruption

Haloacetonitriles are emerging disinfection by-products that can be detected in various aquatic environments. They are cytotoxic, genotoxic, mutagenic, and tumorigenic in vitro and in vivo, but their endocrine-disrupting potency remains unknown. In this study, we examined the agonistic and antagonistic estrogenic and androgenic activities of haloacetonitriles, as well as their cytotoxicity, using a yeast-based reporter assay. We also investigated the interactions of haloacetonitriles with human estrogen receptor alpha (hERα) through molecular docking. We observed that iodoacetonitrile (median lethal dose: 1.96 × 10^-5 M) and bromoacetonitrile (median lethal dose: 1.97 × 10^-5 M) had similar cytotoxicities, which are higher than that of chloroacetonitrile (median lethal dose: 7.16 × 10^-5 M). We observed bromoacetonitrile and chloroacetonitrile elicited estrogenic activity with 10% effective concentrations of 3.30 × 10^-9 M and 2.36 × 10^-9 M, respectively. This finding indicates that bromoacetonitrile and chloroacetonitrile may mimic estrogen signaling through interaction with hERα. Consistent with that result, we identified bromoacetonitrile and chloroacetonitrile interacted with residues in the original estrogen recognition sites of hERα. Our results show that bromoacetonitrile and chloroacetonitrile affect the endocrine-disrupting potency mediated via estrogen receptors by using in vitro assay and molecular docking.