Downregulation of muscarinic receptors gene expression in human breast cancer cellsregulates anchorage-independent cell growthin vitro and angiogenesis in vivo.

NOCYZE, ALEJANDRA; LOMBARDI, GABRIELA; GIAMBALVO GÓMEZ, DILEYVIC; LASERNA, LUCAS NAHUEL; MARTÍNEZ PÉREZ A.M; OROÑO, MANUEL

Mar del plata, Bs. As.

Congreso; Reunión Anual Sociedad Argentina de Investigación Clínica; 2019

SAIC

Downregulation of muscarinic receptors gene expression in human breast cancer cellsregulates anchorage-independent cell growthin vitro and angiogenesis in vivo.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 thathuman breast cancerMCF-7 cells express muscarinic receptors M3 and M4 subtypes and its activation promotes tumoral progression. We previously reported thatthe silencing of both M3 and M4in MCF-7 cells significantly reduced neovascularization capacity of tumoral cellsin vivo.The aim of this work was to evaluate the specific contribution of each M receptor on different tumoral progression parameters likeanchorage-independent cell growth and angiogenesis in vivo. Here, we silencedM3 or M4 subtypes in MCF-7 cells by specific RNAi. After 5 days we used the different experimental groups(siM3, siM4andMCF-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 thatcholinergic stimulation of siMcells showed a significant reduction in colony number when compared withMCF-7+Carb, however this effect was greater in siM3 cells than in siM4 cells (siM3: 99.97±9.50%, siM4: 289.4±5.3% vsMCF-7: 509.1±11.8%;p