TNF-α inhibits trophoblast integration into endothelial cellular networks

Preeclampsia has been linked to shallow trophoblast invasion and failure of uterine spiral artery transformation. Interaction between trophoblast cells and maternal uterine endothelium is critically important for this remodelling. The aim of our study was to investigate the effect of TNF-alpha on the interactions of trophoblast-derived JEG-3 cells into capillary-like cellular networks. We have employed an in vitro trophoblast-endothelial cell co-culture model to quantify trophoblast integration into endothelial cellular networks and to investigate the effects of TNF-alpha. Controlled co-cultures were also treated with anti-TNF-alpha antibody (5 mu g/ml) to specifically block the effect of TNF-alpha. The invasion was evaluated by performing quantitative PCR (Q-PCR) to analyse gene expression of matrix metalloproteinases-2 (MMP-2), MMP-9, tissue inhibitor of matrix metalloproteinase (TIMP)-1, integrins (alpha(1)beta(1) and alpha(6)beta(4)). plasminogen activator inhibitor (PAI)-1, E-cadherin and VE-cadherin. JEG-3 cell integration into endothelial networks was significantly inhibited by exogenous TNF-alpha. The inhibition was observed in the range of 0.2-5 ng/ml, to a maximum 56% inhibition at the highest concentration. This inhibition was reversed by anti-TNF-alpha antibody. Q-PCR analysis showed that mRNA expression of integrins alpha(1)beta(1) and MMP-2 was significantly decreased. VE-cadherin mRNA expression was significantly up-regulated (32-80%, p < 0.01) but its protein concentration in the cell lysates was significantly reduced (20-45%, p < 0.01). PAI-1, MMP-9, TIMP-1 and E-cadherin were not affected. In conclusion. TNF-alpha can inhibit trophoblast-like cells (JEG-3) integration into maternal endothelial cellular networks, and this process involves the inhibition of MMP-2 and a failure of integrins switch from alpha(6)beta(4) to alpha(1)beta(1). These molecular correlations reflect the changes identified in human preeclampsia. (C) 2011 Elsevier Ltd. All rights reserved.