INTERACTION OF NOTCH AND IGF-1 PATHWAYS IN THE MAC-T BOVINE MAMMARY EPITHELIAL CELLS

PERRONE, S; BECÚ-VILLALOBOS, D.; CHIMENTO, A; CRISTINA, C.; BONADEO, NADIA; DALLARD, B.; ISABEL MARIA LACAU

Mar del Plata

Congreso; REUNIÓN ANUAL DE SOCIEDADES DE BIOCIENCIA -XXI Annual Meeting of the Argentinean Biology Society (SAB); 2019

Sociedad Argentina de Biologia

INTERACTION OF NOTCH AND IGF-1 PATHWAYS IN THE MAC-T BOVINE MAMMARY EPITHELIALCELLSWe previously described the involvement of IGF-1 system as well as the expression and activation of the Notch pathway in the pubertal development of the heifers? mammary gland, with a possible role of Notch in cell proliferation and angiogenesis. We therefore aimed to study the effect of Notch pathway inhibition with DAPT, and the IGF-1 treatment on the MAC-T bovine mammary epithelial cell line, in an attempt to elucidate those pathways interaction.Cells were cultured with 50 M DAPT to inhibit Notch pathway during 24, 48 or 72 h, or with DMSO as control. We determined that 72 h DAPT treatment decreases the NOTCH4 receptor expression by Western Blot (p=0,047), and triggers a trend to decrease the HES1 target gene expression (p=0,08), indicating the effectiveness of DAPT in Notch pathway inhibition.We then performed MTS and wound healing assays to functionally study the effect of DAPT on MAC-T cells. The viability of MAC-T cells decreased with 72 h DAPT treatment (p=0,02) and cell migration was altered with 24 h DAPT. Interestingly, mRNA IGFR1 expression increased with 72 h DAPT treatment by RT-qPCR (p=0,03).When MAC-T cells were stimulated with 10 ng/ml IGF-1 during 24 h we observed anincrement in cell viability by MTS assay and cell migration was altered respect to the control, determined by wound healing assay. In addition, we observed a trend to increase the expression of the Notch target gene HEY1 with 10 ng/ml IGF-1 treatment, by RT-qPCR.To study IGF-1 and Notch pathway interaction we treated the MAC-T cells with 50 MDAPT during 24, 48 and 72 h and 10 ng/ml IGF-1 during 24 or 48 h. We observed that IGF1 reversed the decrease in cell viability induced by DAPT (72h DAPT-24h IGF-1).These results suggest that IGF-1 and Notch pathways may interact to regulate cell proliferation and migration in the bovine mammary cells. Further studies are needed to deepen the knowledge of the regulation of mammary gland development.