Evaluation of muscarinic receptors gene editing by CRISPR/cas9 on migration, spheroidgrowth and angiogenesis in human breast cancer cells.

NOCYZE, ALEJANDRA; MARTINEZ, ADRIANA MARCELA; ORABONA, MARÍA AGUSTINA; GIAMBALVO GÓMEZ, DILEYVIC; LOMBARDI, GABRIELA

Mar del plata, Bs. As.

Congreso; Reunión Anual Sociedad Argentina de Investigación Clínica; 2019

SAIC

Evaluation of muscarinic receptors gene editing by CRISPR/cas9 on migration, spheroidgrowth and angiogenesis in human breast cancer cells.Martínez Pérez A.M., Giambalvo Gómez D., Nozyce A., Orabona A., Lombardi M.G.Laboratorio de Oncoinmunología Molecular.Centro de Estudios Farmacológicos y Botánicos (CEFyBO) UBA-CONICET.It has been reported that muscarinic receptors (M) are absent in normal breast cells and are up-regulated in tumor cells. Particularly, human breast cancer MCF-7 cellsexpress M3 and M4subtypes andits activation promotes cancer progression. We demonstrated that M expression in non-tumorigenic human mammary cell lines triggers malignant transformation. To confirm the contribution of M ontumoral progression, we developed new cell lines by genome editing of M3 and/or M4 receptors in MCF-7 cells using specific CRISPR-Cas9-gRNA complexes(cM3, cM4 and cM3M4). We analyzed the effect on cell migration capacityby wound healing assay, on the ability to generate three-dimensional structures (spheroids)in vitroand on induced angiogenesis in vivo. All cM cell lines significantly decreased their migration capacity (cM3:67+2%; cM4: 77+0.4%; cM3/4:36.5+6.7%; n=3)in comparison with MCF-7 cells(considered as 100%, p