Glyphosate concentration detected in human breast milk alters milk proteins level in a mouse mammary cell line

ROTH IL; ODDI SL; GOMEZ AL; SCHIERANO-MAROTTI G; KASS L; ALTAMIRANO GA

Mar del Plata

Congreso; REUNIÓN CONJUNTA SAIC SAB AAFE AACYTAL 2023; 2023

Previously, we demonstrated that milk proteinsynthesis is affected by environmental estrogens using a 3D in vitro model. Inrecent years, the broad-spectrum herbicide glyphosate (GLY) has been classifiedas a potential estrogenic and androgenic compound. Here, our aim was to analyzewhether direct exposure to glyphosate-based herbicide (GBH) and its active ingredientGLY affects milk proteins synthesis during mammary differentiation using the murinecell line HC11. Cells were grown in medium supplemented with epidermal growthfactor, insulin and fetal bovine serum (FBS) until confluent. Cells were then culturedfor 72 h with medium supplemented with 2% charcoal-stripped FBS, after whichthey were differentiated with lactogenic hormones (prolactin, insulin anddexamethasone) and exposed for 72 h to: a) Vehicle (phenol red-free medium), b)0.01 and 0.5 μM GBH and c) 0.01 and 0.5 μM GLY. The concentrationsof GBH and GLY were selected considering the detected values of GLY both in humanmilk and in serum and urine of pregnant women. After 72 h ofdifferentiation and exposure to GBH and GLY, cell viability assay was carriedout using the WST-1 kit. Also, the mRNA levels of milk proteins: beta-casein (CSN2)and whey acidic protein (WAP), considered markers of functionaldifferentiation, were analyzed by real-time RT-PCR. No significant differencesin cell viability were found between the treatments. On the other hand, themRNA level of CSN2 was increased in HC11 cells treated with 0.01 µM GBH and GLY(p<0.05). In addition, the WAP mRNA level was only increased in cellstreated with 0.01 µM GBH. These findings show that direct exposure to aconcentration similar to that found in breast milk (0.01 μM) alters milkprotein synthesis. In conclusion, exposure to the herbicide could interferewith the mammary functional differentiation and, therefore, with the growth andhealth of the offspring.