EXPLORING GENISTEIN’S IMPACT ON CELL ADHESION IN BOVINE OVIDUCTAL EPITHELIAL CELLS

VELLA, MILDA A.; DE BOECK, MAXIMILIANO; GARCÍA, DANIELA C.; VALDECANTOS, PABLO A; MARIELA ROLDÁN OLARTE

Mar del Plata

Congreso; Reunión Conjunta de Sociedades de Biología; 2023

SAIC/ SAB/ AAFE7 AACYTAL

An optimal oviductal microenvironment is pivotal during the early phases of the reproductive process in mammals. The adhesion of cells that constitutes the oviductal epithelium is of importance to contribute to the maintenance of structural integrity and the efficiency of gamete transport, sperm capacitation and ensures the proper exchange of nutrients between the nterstitial cellular space and the oviductal lumen. Genistein (GNT), an isoflavone with multiple biological properties, is capable of diffusing through the oviductal epithelium. Given the significance of preserving oviductal integrity and the diverse properties of GNT, the aim of this work was to evaluate its impact on cell adhesion in bovine oviductal epithelial cells (BOECs). Cells were obtained for mechanical pressure from recently slaughtered heifer oviducts. Explant and monolayer cultures were stimulated with GNT at various concentrations (0.1 µM - 10 µM) for 24 hours. Subsequently, BOEC adhesion was assessed. To achieve this, monolayer cultures were dissociated with 0.25% trypsin, and 10000 live cells were seeded in wells previously coated with type I collagen, derived from dissected bovine tendons. After 2 hours, cells were washed with PBS solution to remove debris and unattached cells; then were stained with 0.1% Crystal violet for adherent cell visualization and count them with ImageJ software. A decrease in the number of adherent BOECs per mm² was observed with all tested GNT concentrations, highlighting that with 10 µM the most pronounced effect was recorded. The in vitro expression of PTK2 and PXN, genes involved in focal adhesion formation, was evaluated using RT-PCR. In GNT- treated cultures, a reduction in the expression of both genes was evident. These findings lead us to conclude that GNT-treated cultures showed reduced expression of both genes, suggesting that this isoflavone negatively impacts in BOEC adhesion in a dose-dependent manner, possibly by influencing focal adhesion-related genes.