Glyphosate-based herbicides are the most widely used across the world; they are commercialized
in different formulations. Their residues are frequent pollutants in the environment.
In addition, these herbicides are spread on most eaten transgenic plants, modified
to tolerate high levels of these compounds in their cells. Up to 400 ppm of their
residues are accepted in some feed. We exposed human liver HepG2 cells, a well-known
model to study xenobiotic toxicity, to four different formulations and to glyphosate,
which is usually tested alone in chronic in vivo regulatory studies. We measured cytotoxicity
with three assays (Alamar Blue, MTT, ToxiLight), plus genotoxicity (comet assay),
anti-estrogenic (on ERalpha, ERbeta) and anti-androgenic effects (on AR) using gene
reporter tests. We also checked androgen to estrogen conversion by aromatase activity
and mRNA. All parameters were disrupted at sub-agricultural doses with all formulations
within 24h. These effects were more dependent on the formulation than on the glyphosate
concentration. First, we observed a human cell endocrine disruption from 0.5 ppm on
the androgen receptor in MDA-MB453-kb2 cells for the most active formulation (R400),
then from 2 ppm the transcriptional activities on both estrogen receptors were also
inhibited on HepG2. Aromatase transcription and activity were disrupted from 10 ppm.
Cytotoxic effects started at 10 ppm with Alamar Blue assay (the most sensitive), and
DNA damages at 5 ppm. A real cell impact of glyphosate-based herbicides residues in
food, feed or in the environment has thus to be considered, and their classifications
as carcinogens/mutagens/reprotoxics is discussed.