title: Stimulation of the JAK/STAT pathway by LIF and OSM in the human granulosa cell line COV434.
abstract: The development of the follicle and competent oocyte is highly coordinated, requiring interplay among several systems. These implicate endocrine, immune, and metabolic signals, intrafollicular paracrine factors from theca, mural, and cumulus granulosa cells, and the oocyte itself. Granulosa cells play a key role in their interaction. COV434 is one of the few human granulosa cell lines that can be used as an in vitro model for ovarian research. We aimed to evaluate the possible activation of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway by IL-6-type cytokines leukemia inhibitory factor (LIF) and oncostatin M (OSM) in COV434 cells. Expression of GP130 (glycoprotein 130), STAT3 (signal transducer and activators of transcription 3), PIAS3 (protein inhibitor of activated STAT 3), and SOCS3 (suppressor of cytokine signaling 3) genes after stimulation with LIF or OSM was assessed using RT-qPCR (real-time PCR). GP130 transcripts were significantly upregulated after incubation with LIF or OSM for 24h. Expression of the STAT3 gene was stimulated only after incubation with LIF, but not OSM. SOCS3 showed significant upregulation for all time periods and the levels of PIAS3 were initially down- and after 24h upregulated. Furthermore, the major signaling components of the JAK/STAT pathway, GP130 and STAT3, and the kinase activation patterns of STAT3, were examined at protein level. We found constitutive protein expression for GP130, STAT3, pSTAT3(ser727) and upregulation of pSTAT3(tyr705) by LIF and OSM. Our results demonstrate the activation of the JAK/STAT pathway by LIF and OSM in human granulosa cells.
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany.
authors: Jana
authors: J
authors: Pastuschek
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany; Institute of Immunology, Hannover Medical School, Hannover, Germany.
authors: Jenny
authors: J
authors: Poetzsch
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany.
authors: Diana M
authors: DM
authors: Morales-Prieto
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany.
authors: Ekkehard
authors: E
authors: Schleußner
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany. Electronic address: markert@med.uni-jena.de.
authors: Udo R
authors: UR
authors: Markert
authors: Placenta-Lab, Department of Obstetrics, University Hospital Jena, Germany; Department of Cytology, Histology and Embryology, Biology Faculty, Sofia University, Sofia, Bulgaria.
authors: Georgi
authors: G
authors: Georgiev
mesh_terms: D002450:Cell Communication; D002460:Cell Line; D050822:Cytokine Receptor gp130 / Q000235:genetics / Q000378:metabolism*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006107:Granulosa Cells / Q000276:immunology*; D006801:Humans; D053612:Janus Kinases / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000276:immunology; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D053683:Oncostatin M / Q000276:immunology; D011247:Pregnancy; D050828:Protein Inhibitors of Activated STAT / Q000235:genetics / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D053605:Leukemia Inhibitory Factor; D018832:Molecular Chaperones; C484745:PIAS3 protein, human; D050828:Protein Inhibitors of Activated STAT; C106644:SOCS3 protein, human; D050796:STAT3 Transcription Factor; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D053683:Oncostatin M; D050822:Cytokine Receptor gp130; D053612:Janus Kinases
keywords: COV434 cell line; Granulosa cells; IL6-type cytokines; JAK/STAT pathway; LIF; OSM