Shiga toxin type 2 suppresses human extravillous trophoblast migration and invasion

SCALISE M.LUJAN; GARCIA CECILIA; DAMIANO ALICIA; SACERDOTI FLAVIA; IBARRA CRISTINA

CABA

Congreso; Reunión conjunta de sociedades de biociencias; 2017

LXII Reunión de la Sociedad Argentina de Investigación Clínica (SAIC).

During human placentation, the extravillous trophoblast (EVT) invades the maternal decidua and reconstructs maternal spiral arteries. We have previously reported that Shiga toxin producing Escherichia coli (STEC) infections during pregnancy may cause maternal or fetal damage mediated by Shiga toxin type 2 (Stx2). We propose that an early STEC infection during pregnancy could damage human trophoblast mediated by Stx2. The aim of this work was to evaluate the effects of Stx2 on cell viability, migration and invasion of human EVT. Swan 71 cell line was used as human first trimester trophoblast model. Firstly, cells were submitted to a lipid extraction for globotriaosylceramide (Gb3, Stx2 receptor) analysis by thin layer chromatography. Cell viability was evaluated by neutral red uptake at 24, 48 and 72h of treatment with Stx2 (0.001-1μg/ml). To assess the rates of apoptosis, cells were fluorescently stained with orange acridine/ethidium-bromide and analyzed under optical microscopy. The effects of Stx2 on migration were studied by the wound-healing assay at 8 and 24h. Proteine activity of matrix metaloproteinase-2 (MMP-2) and invasiveness were analyzed at 24h after treatment by zymography and Matrigel invasion assay, respectively. Gb3 was expressed in Swan 71. Stx2 (1μg/ml) inhibited cell viability and increased the apoptotic rate at 72h respectively (p