Muscarinic receptors in non-tumorigenicMCF-10A breast cells. effect of carbachol on three-dimensional growth and vegf-A expression

LOMBARDI, GABRIELA; SALES, MARIA ELENA; MARTINEZ PULIDO, PAOLA

CABA

Congreso; Reunión Conjunta de Sociedades de Biomedicina,; 2017

9 Sociedades de Biomedicina

It was reported that muscarinic acetylcholine receptors (mAChRs)are constitutively expressed in different types of tumors. In our laboratorywe demonstrated that the activation of mAChRs with thecholinergic agonist carbachol (CARB) promotes malignant progressionin murine and human breast tumors. In line with theseresults, we developed a new model to confirm the contribution ofmAChRs to malignant transformation, transfecting the non-tumorigenichuman mammary cell line, MCF-10A with mAChRs. Consideringthat bidimensional culturing is the most used techniquebut not the best model to mimic the first stages of tumor growthin vivo, we developed three-dimensional (3D) cultures (spheroids)with mAChRs-transfected MCF-10A cells and analyzed the effect ofmAChRs activation on growth (ìm2) and the expression of vascularendothelial growth factor-A (VEGF-A) by Western blot (%). Thede novo expression of mAChRs in MCF-10A cells promoted theformation of spheroids an VEGF-A expression in 3D cultures. Theaddition of CARB increased spheroids size (mAChR3:4745±502;mAChR4: 5427±357; mAChR3R4: 6055±475) in comparison to untreatedcells (mAChR3:3350±467; mAChR4: 2903±462; mAChR3R4:4330±362)(p