Downregulation of muscarinic receptors gene expression in human breast cancer cells regulates anchorage-independent cell growth in vitro and angiogenesis in vivo.

NOZYCE ALEJANDRA; LOMBARDI, GABRIELA; GIAMBALVO, DILEYVIC; LASERNA LUCAS; MARTINEZ ADRIANA; ORONO, MANUEL

Buenos Aires

Congreso; Reunión anual de sociedades de biociencia 2019; 2019

Downregulation of muscarinic receptors gene expression in human breast cancer cells regulates anchorage-independent cell growth in vitro and angiogenesis in vivo.Giambalvo Dileyvic, Martínez Adriana, Alejandra Nozyce, Laserna Lucas, Oroño Manuel, Lombardi María Gabriela.Laboratorio de Oncoinmunología Molecular.Centro de Estudios Farmacológicos y Botánicos (CEFyBO) UBA-CONICET.Muscarinic receptors (M) expression, activation and signalling play important roles in regulating many cellular process and cancer progression. It has been reported that human breast cancer MCF-7 cells express muscarinic receptors M3 and M4 subtypes and its activation promotes tumoral progression. We previously reported that the silencing of both M3 and M4 in MCF-7 cells significantly reduced neovascularization capacity of tumoral cells in vivo. The aim of this work was to evaluate the specific contribution of each M receptor on different tumoral progression parameters like anchorage-independent cell growth and angiogenesis in vivo. Here, we silenced M3 or M4 subtypes in MCF-7 cells by specific RNAi. After 5 days we used the different experimental groups (siM3, siM4 and MCF-7 cells with and without carbachol (Carb, -8M)) in the following assays. Briefly, for soft agar colony assay we seeded 2X104 cells of each group into medium with soft agar. After 2 weeks, the colonies larger than 60µm in diameter were counted. We observed that cholinergic stimulation of siM cells showed a significant reduction in colony number when compared with MCF-7+Carb, however this effect was greater in siM3 cells than in siM4 cells (siM3: 99.97±9.50%, siM4: 289.4±5.3% vs MCF-7: 509.1±11.8%; p