title: Decidual NK cell-derived conditioned medium enhances capillary tube and network organization in an extravillous cytotrophoblast cell line.
abstract: Extravillous cytotrophoblast (EVT) migration, invasion and endovascular differentiation are regulated by a variety of growth factors, cytokines and adhesion molecules. Decidual natural killer cells (dNK) and their secreted cytokines probably modulate these processes. In this study, we used dNK-derived conditioned medium (dNK-CM) to investigate whether or not (i) dNK-CM was able to enhance capillary tube and network formation of an EVT cell line, HTR8/SVneo, on Matrigel, (ii) PI3K/AKT pathway and p38 MAPK pathway activation were involved, and (iii) HTR8/SVneo surface ICAM-1 played a role in the process of HTR8/SVneo endovascular differentiation. The results demonstrated that HTR8/SVneo constitutively form 'vascular' tubes and networks after culture on Matrigel. dNK-CM enhanced and maintained tube and network formation, acquiring an endothelium-like angiogenic morphology followed by increased VEGF-C production. HTR8/SVneo cell expression level of VE-cadherin, PECAM-1, VCAM-1 and alphavbeta3 was unaltered by dNK-CM, whereas ICAM-1 expression level was increased. Anti-human ICAM-1 blocking antibody inhibited HTR8/SVneo migration and partially reversed dNK-CM-mediated enhancement of HTR8/SVneo tube and network formation. PI3K/AKT and p38 MAPK pathways were activated in dNK-CM-mediated enhancement of HTR8/SVneo tube and network formation. The PI3K/AKT and p38 MAPK pathway inhibitors (LY294002 and SB202190, respectively) decreased dNK-CM-stimulated ICAM-1 induction, HTR8/SVneo migration, and reversed tube and network formation. The results suggest that dNK cell-secreted growth factors and cytokines participate in the regulation of HTR8/SVneo endothelium-like tube formation. Adhesion molecules, particularly ICAM-1, expressed on EVT may participate in the process. To our knowledge, this is the first report of a role for ICAM-1 in EVT angiogenesis, as previously reported for endothelial cells.
authors.affiliation: Department of Obstetrics and Gynaecology, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Y
authors.initials: Y
authors.lastname: Hu
authors.forename: G
authors.initials: G
authors.lastname: Eastabrook
authors.forename: R
authors.initials: R
authors.lastname: Tan
authors.forename: C D
authors.initials: CD
authors.lastname: MacCalman
authors.forename: J P
authors.initials: JP
authors.lastname: Dutz
authors.forename: P
authors.initials: P
authors.lastname: von Dadelszen
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002867:Chromones / Q000494:pharmacology; D017077:Culture Media, Conditioned; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000502:physiology*; D007694:Killer Cells, Natural / Q000502:physiology*; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D011247:Pregnancy; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000502:physiology; D011725:Pyridines / Q000494:pharmacology; D018709:Statistics, Nonparametric; D014327:Trophoblasts / Q000201:enzymology / Q000502:physiology*; D042582:Vascular Endothelial Growth Factor C / Q000502:physiology*
chemical_list: D002867:Chromones; D017077:Culture Media, Conditioned; D007093:Imidazoles; D009025:Morpholines; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D042582:Vascular Endothelial Growth Factor C; D018799:Intercellular Adhesion Molecule-1; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole