CEFYBO   02669
CENTRO DE ESTUDIOS FARMACOLOGICOS Y BOTANICOS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Muscarinic receptors expressed in non-tumorigenic MCF-10A cells induced a malignant phenotype that activates human endothelial cells.
Autor/es:
MARTÍNEZ, P PULIDO; FIZMAN, GABRIEL; LOMBARDI, GABRIELA; SALES, M E; SANCHEZ FRANCISCO
Lugar:
Mar del Plata
Reunión:
Congreso; LXIII Reunión Anual de la Sociedad Argentina de Investigación Clínica-LXVI Reunión Anual de la Sociedad Argentina de Inmunología-Reunión Anual de la Sociedad Argentina de Fisiología; 2018
Institución organizadora:
SAIC SAI SAFIS
Resumen:
Our laboratory reported the participation of muscarinic acetylcholine receptors (mAChR) in different steps of tumor progression in murine and human breast cancer. To confirm the contribution of mAChR to malignant transformation, we developed a new cell line by stably transfecting the non-tumorigenic human mammary cell line MCF-10A with mAChR, subtypes 3 and/or 4. Transfected cells acquired the ability to generate three-dimensional structures (spheroids) that mimic the first stages of tumor growth in vivo. Tumor microenvironment has a crucial role in tumorigenesis, particularly endothelial cells, as they form the tumor microvasculature. Considering the later, we analyzed the effect of spheroid-derived conditioned media (S-DCM) from different days of culture on human mammary endothelial cells-1 (HMEC-1) viability, vascular endothelial growth factor-A (VEGF-A) expression and tubulogenesis. S-DCM from 14 days significantly increased HMEC-1 cell viability (mAChR3:153.8%+11,7; mAChR4: 211.2+13.3%; mAChR3/4:171.9+13.5%; n=3) in comparison with conditioned media from MCF-10A (100+18.6%), and in a similar manner than MCF-7 tumor cells (155.6+16.7%; n=3). VEGF-A expression was up-regulated when HMEC-1 cells were treated with S-DCM of 14 days (mAChR3:167.4+5.6%; mAChR4:61.5+10.7%; mAChR3R4:126.7+7.7%; n=3) comparably to S-DCM from MCF-7 tumor cell spheroids (109.5+23.0; n=3). Finally, we investigate the effect of S-DCM of 14 days on HMEC-1 cells in a tube formation assay (mAChR3:119.4+9.9; mAChR4: 117.5+9.8; mAChR3R4:102.2+7.2; MCF-7:130.6+6.33) vs. HMEC-1 cells without treatment. In conclusion, the transfection of non-tumorigenic breast cells MCF-10A with mAChR induces a malignant phenotype that triggers three-dimensional growth and the liberation of pro-angiogenic mediators stimulating HMEC-1 cell viability, VEGF-A expression and tubulogenesis in a similar manner to that observed in breast tumors.