Leptin is involved in human trophoblastic migration and invasion

AYELEN TORO; SAMPAYO, ROCÍO; PEREZ PEREZ, ANTONIO; MASKIN, BERNARDO; SIMIAN, MARINA; SÁNCHEZ MARGALET, VICTOR; CECILIA VARONE

Mar del Plata

Congreso; Latin American Symposium on Maternal Fetal Interaction & Placenta; 2015

Cytotrophoblastic (CTB) cells of the human placenta proliferate, migrate and invade the uterus during pregnancy to allow implantation and placentation. To successfully become invasive the CTB cells undergo a transition from epithelial to highly invasive mesenchymal phenotype, called EMT (epithelial-mesenchymal transition). EMT is characterized by up-regulation of motility promoting molecules such as the adhesion molecule β1 Integrin and the loss of epithelial markers such as E-cadherin. Leptin is produced by placenta and we have previously demonstrated that promotes proliferation and survival of trophoblastic cells. OBJECTIVEWe aimed to study a possible role of leptin in cell migration and invasion by evaluating its effect on β1 Integrin and E-cadherin expression. METHODSThe human CTB cell line from first trimester Swan-71 was used. Cells were treated with different leptin concentrations (50, 100 and 250 ng/ml) for 48h. To study if leptin effect was reversible, after leptin treatment, media with leptin was replaced with fresh media for 24h. We also investigated if metalloproteinases (MMPs) were necessary for leptin action. For this aim, cells were cultured with a MMPs inhibitor (GM6001, 10µM) during 2h before leptin treatment. β1 Integrin and E-cadherin expression were analyzed by qRT-PCR or/and WB assays. To evaluate leptin effect on cell migration wound healing assays were performed.RESULTSLeptin significantly reduced E-cadherin expression at mRNA level and in a dose dependent manner at protein level. This effect was reversible and MMPs activity was necessary. β1 Integrin protein levels were ̴ 2 fold increased after stimulation with leptin (p