title: TNF-α inhibits trophoblast integration into endothelial cellular networks.
abstract: 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-α 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-α. Controlled co-cultures were also treated with anti-TNF-α antibody (5 μg/ml) to specifically block the effect of TNF-α. 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 (α(1)β(1) and α(6)β(4)), plasminogen activator inhibitor (PAI)-1, E-cadherin and VE-cadherin. JEG-3 cell integration into endothelial networks was significantly inhibited by exogenous TNF-α. 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-α antibody. Q-PCR analysis showed that mRNA expression of integrins α(1)β(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-α 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 α(6)β(4) to α(1)β(1.) These molecular correlations reflect the changes identified in human preeclampsia.
authors.affiliation: Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Australia. xub@hri.org.au
authors.forename: B
authors.initials: B
authors.lastname: Xu
authors.forename: S
authors.initials: S
authors.lastname: Nakhla
authors.forename: A
authors.initials: A
authors.lastname: Makris
authors.forename: A
authors.initials: A
authors.lastname: Hennessy
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D015820:Cadherins / Q000235:genetics / Q000276:immunology; D002450:Cell Communication / Q000187:drug effects* / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D018920:Coculture Techniques; D005260:Female; D006801:Humans; D016023:Integrins / Q000235:genetics / Q000276:immunology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000276:immunology; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000276:immunology; D009215:Myometrium / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000276:immunology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000276:immunology; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D016023:Integrins; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D017395:Plasminogen Activator Inhibitor 1; D012313:RNA; D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D000099152:Cadherin 5