THE DISRUPTION OF CAVEOLAE/CAVEOLIN-1 PRESENT IN TROPHOBLAST CELLS DID NOT AFFECT ENDOTHELIAL CELL MIGRATION

SZPILBARG NATALIA; REPPETTI, JULIETA; SEYAHIAN, ABRIL; MARTINEZ, NORA; DAMIANO, ALICIA E; RECA, ALEJANDRA

Chillán

Congreso; II MEETING IN RESEARCH AND INNOVATION IN VASCULAR HEALTH, IV RESEARCH MEETING IN HYPERTENSION IN PREGNANCY; 2017

Migration plays key roles in many physiological and pathological processes including development, angiogenesis, tissue regeneration and metastasis. Caveolin-1 (Cav-1) and caveolae constitute domains that compartmentalize intracellular signalling pathways. During placentation the human extravillous trophoblast (EVT) cells invade the endometrium and maternal vasculature within the uterus. Objective: To evaluate the role of caveolae/Cav-1 in the migration and invasion of trophoblast cells and the interaction with the endothelial cells.Methods: Cav-1 expression was examined in trophoblast cells Swan-71 by immunocytochemistry and western blot. Cells were treated with 10 mM methyl-β-cyclodextrin (β-MCD) to disrupt caveolae. Conditioned medium (CM) was also collected. Migration was assessed by wound healing assay at 8, 18 and 24 h. To evaluate invasion, trophoblast cells were seeded in the top of transwell inserts of 8-µm pore precoated with Matrigel (0.3 mg/ml). After 24 h, cells located on the undersurface were fixed with methanol and stained with hematoxylin. To evaluate the interaction with the endothelial cells (EA.hy926), proliferation (evaluated by MTT assay) and migration were performed after the culture of the cells with the CM obtained from Swan-71 cells.Results: The migration of Swan-71 treated with β-MCD was decreased compared with control: 8 h (32.78 ± 1.14 vs 17.78 ± 0.5561), 18 h (66.74 ± 1.434 vs 51.79 ± 1.158), 24 h (84.45 ± 1.160 vs 64.95 ± 0.7165). In the Invasion assay in Swan-71 no differences were observed between control and β-MCD treated cells. No differences were also found in the proliferation and in the migration of endothelial cells treated with the CM from Swan-71.Conclusion: These results show that lipid raft disruption significantly attenuates migration (but no invasion) in Swan-71 cells suggesting a role for Caveolae/caveolin-1 during the early stages of placental development. Nevertheless the lack of caveolae/caveolin-1 in the trophoblast cells did not affect the proliferation and the migration of endothelial cells.