IBYME   02675
INSTITUTO DE BIOLOGIA Y MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
NOTCH PATHWAY ACTIVATION IN MAC-T BOVINE MAMMARY EPITHELIAL CELLS
Autor/es:
SOFÍA PERRONE; LACAU ISABEL MARÍA; DAMASIA BECU-VILLALOBOS; NADIA BONADEO; CAROLINA CRISTINA
Lugar:
Buenos Aires
Reunión:
Congreso; Reunión Conjunta de Sociedades de Biociencias; 2017
Institución organizadora:
Sociedad Argentina de Investigación Clínica SAIC
Resumen:
We previously described 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. In this work, we aimed to study basal expression and the prolactin regulation of the Notch pathway components in the MAC-T bovine mammary epithelial cell line. MAC-T cells were grown in 50% DMEM and 50% RPMI medium containing 10% fetal bovine serum (FBS), 5 μg/mL insulin, 5 μg/mL hydrocortisone, 1% glutamine and 1% penicillin/streptomycin at 37°C in an atmosphere of 5% CO2. They were plated at a density of 45,000 cells/well and cultured in 5% FBS as control. Treated cells were cultured with 1 or 5 μg/mL prolactin during 96 h. Proteins were extracted to perform Western Blot assay and mRNA was isolated for Real Time PCR. Notch receptors in its active (80 KDa) and membrane (110 KDa) forms, target gene Hes1 and p-ERK expression levels were determined by Western blot and Jagged1 ligand and the Hes1 and Hey1 target genes were evaluated by qRT-PCR. We determined that MAC-T cells express several components of the pathway: the four Notch receptor isoforms, the Jagged1 ligand and the Hes1 and Hey1 target genes. We found a dose dependent increase in p-ERK expression (p=0.04) and a trend to increase in Notch2 and Notch4 active domains with prolactin treatment. It was described that ERK1/2 phosphorylation is upstream to the Jagged1 gene induction and that it is increased by prolactin in cancer cells. Our results suggest that prolactin would induce Notch pathway activation in these bovine mammary cells