title: Oncostatin M and leukaemia inhibitory factor trigger signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 pathways but result in heterogeneous cellular responses in trophoblast cells.
abstract: Leukaemia inhibitory factor (LIF) and oncostatin M (OSM) are pleiotropic cytokines present at the implantation site that are important for the normal development of human pregnancy. These cytokines share the cell membrane receptor subunit gp130, resulting in similar functions. The aim of this study was to compare the response to LIF and OSM in several trophoblast models with particular regard to intracellular mechanisms and invasion. Four trophoblast cell lines with different characteristics were used: HTR-8/SVneo, JEG-3, ACH-3P and AC1-M59 cells. Cells were incubated with LIF, OSM (both at 10ngmL(-1)) and the signal transducer and activator of transcription (STAT) 3 inhibitor S3I-201 (200µM). Expression and phosphorylation of STAT3 (tyr(705)) and extracellular regulated kinase (ERK) 1/2 (thr(202/204)) and the STAT3 DNA-binding capacity were analysed by Western blotting and DNA-binding assays, respectively. Cell viability and invasiveness were assessed by the methylthiazole tetrazolium salt (MTS) and Matrigel assays. Enzymatic activity of matrix metalloproteinase (MMP)-2 and MMP-9 was investigated by zymography. OSM and LIF triggered phosphorylation of STAT3 and ERK1/2, followed by a significant increase in STAT3 DNA-binding activity in all tested cell lines. Stimulation with LIF but not OSM significantly enhanced invasion of ACH-3P and JEG-3 cells, but not HTR-8/SVneo or AC1-M59 cells. Similarly, STAT3 inhibition significantly decreased the invasiveness of only ACH-3P and JEG-3 cells concomitant with decreases in secreted MMP-2 and MMP-9. OSM shares with LIF the capacity to activate ERK1/2 and STAT3 pathways in all cell lines tested, but their resulting effects are dependent on cell type. This suggests that LIF and OSM may partially substitute for each other in case of deficiencies or therapeutic interventions.
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Wittaya
authors.initials: W
authors.lastname: Chaiwangyen
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Ospina-Prieto
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Morales-Prieto
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Francisco Lazaro
authors.initials: FL
authors.lastname: Pereira de Sousa
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Pastuschek
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Justine S
authors.initials: JS
authors.lastname: Fitzgerald
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Schleussner
authors.affiliation: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Bachstrasse 18, 07743 Jena, Germany.
authors.forename: Udo R
authors.initials: UR
authors.lastname: Markert
mesh_terms: D001665:Binding Sites; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D004247:DNA / Q000378:metabolism; D005260:Female; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D053683:Oncostatin M / Q000494:pharmacology*; D010766:Phosphorylation; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014327:Trophoblasts / Q000187:drug effects* / Q000201:enzymology
chemical_list: D053605:Leukemia Inhibitory Factor; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D053683:Oncostatin M; D004247:DNA; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9