- Role of reproductive bioactive lipids in human first trimester trophoblast migration.

CAÑUMIL VA; RIBEIRO ML; BELTRAME JS; MARKERT UR; SORDELLI MS; FRANCHI AM

Orlando

Congreso; 64th Annual Scientific Meeting of the Society of Reproductive Investigation; 2017

Society of Reproductive Investigatiob

Introduction: Bioactive lipids as lysophosphatidic acid(LPA) and prostaglandins (PGs) play major roles at the maternal-fetal interface.We reported that LPA augments cyclooxygenase-2 (COX-2) derived PGE2 synthesisin the rat uterus during implantation. Trophoblasts are the main source of LPA infirst trimester pregnancy, suggesting that LPA participates in its functions. PGE2regulates decidualization and vascularization, two processes parallel totrophoblast migration and invasion. Defects in these events are related toimplantation failures. Thus, we postulate that LPA and PGs crosstalk modulatecrucial events at the maternal-fetal interface. Our aim was to investigate LPAaction on human first trimester trophoblast migration and its interaction with PGE2. Methods: HTR-8/SVneo cell line was used as a model of human firsttrimester extravillous trophoblast. Cells were subjected to migration (woundhealing assay, 18h), invasion (transwell inserts of 8 µm coated with Matrigel,24h) and proliferation assays (MTS, 48h). PGE2 concentration was determined byradioimmunoassay and the localization of COX-1 and 2 by immunocytochemistry. Results: Treatment of HTR-8/SVneo with LPA (50 µM) augments trophoblastmigration and invasion (p<0.05), but does not modify proliferation. LPAconcentration was selected based on the physiological concentration found inplasma of women in the first trimester of gestation. The co-incubation of LPAtogether with BrP-LPA (10-5M, an antagonist of LPA1 to LPA4) revertsLPA effect on trophoblast migration (p<0.05), suggesting that Gprotein-coupled receptors are involved. To investigate the participation of PGsin LPA action, cells were co-incubated with LPA and indomethacin (10-6M,a COX-1 and 2 inhibitor) or NS-398 (10-6M, a selective COX-2 inhibitor).COXs inhibitors diminish LPA increased migration, indicating that COX derived PGsmediate the increase triggered by LPA on trophoblasts cells. Interestingly, LPAstimulates PGE2 production (p<0.05) and HTR-8/SVneo stain positive for COX-1and 2 in the cytoplasm and plasma membrane. Conclusion: Our results suggest that LPAperform primordial functions at the maternal-fetal interface modulatingtrophoblast invasion and migration. LPA stimulates migration by a PGs mediated pathway.Localization of COXs suggests a possible interaction with LPAreceptors during this mechanism.