HYPOXIA INDUCIBLE FACTOR-1 ALPHA IMPAIRS SYNCYTIALIZATION OF HUMAN TROPHOBLAST BEWO CELLS: ROLE OF ENDOCANNABINOID SYSTEM.

ACCIALINI PAULA; SARACO NORA; PALLIGAS MARCOS; MARTINEZ NORA; ETCHEVERRY TOMÁS; ABÁN CYNTIA; FARINA MARIANA

CABA

Congreso; 2019 IFPA - VIII SLIMP; 2019

Objectives: We have reported that hypoxia inducible factor-1 alpha (HIF-1α) deregulates the endocannabinoid system (ECS). The aim of the present work was to assess the impact of HIF-1α stabilization and ECS on BeWo cell line syncytialization.Methods: BeWo cell line was cultured with 25µM forskolin (FSK) to induce cell fusion. Stimulated cells were treated with or without cobalt chloride (50µM-100µM CoCl2), a hypoxia-mimetic agent that stabilizes HIF-1α; or R-(+)-Methanandamide (0.01µM-10µM Met-AEA), a non-hydrolyzable anandamide analogous. AM251, SR141716 and AM630 (0.01µM-1µM) were used to antagonize cannabinoid receptors (CBs). Cell viability and integrity were analyzed by MTT assay and LDH activity, respectively. HIF-1α stabilization was confirmed by Western blot. Fatty acid amide hydrolase (FAAH), the enzyme that degrades anandamide, mRNA levels were measured by RT-qPCR. Several parameters were used to study syncytialization: GCM1 mRNA levels by RT-qPCR; human chorionic gonadotropin (hCG) release by chemiluminescence immunoassay; syncytin-1 protein expression, propidium iodide (PI) staining and cell size by flow cytometry; and E-cadherin distribution by immunocytochemistry.Results: Cells treated with FSK showed increased GCM1 mRNA levels, hCG secretion, syncytin-1 protein expression, DNA content (PI staining) and cell size compared to the non-treated control (p