title: Nitric Oxide Prevents Mouse Embryonic Stem Cell Differentiation Through Regulation of Gene Expression, Cell Signaling, and Control of Cell Proliferation.
abstract: Nitric oxide (NO) delays mouse embryonic stem cell (mESC) differentiation by regulating genes linked to pluripotency and differentiation. Nevertheless, no profound study has been conducted on cell differentiation regulation by this molecule through signaling on essential biological functions. We sought to demonstrate that NO positively regulates the pluripotency transcriptional core, enforcing changes in the chromatin structure, in addition to regulating cell proliferation, and signaling pathways with key roles in stemness. Culturing mESCs with 2 μM of the NO donor diethylenetriamine/NO (DETA/NO) in the absence of leukemia inhibitory factor (LIF) induced significant changes in the expression of 16 genes of the pluripotency transcriptional core. Furthermore, treatment with DETA/NO resulted in a high occupancy of activating H3K4me3 at the Oct4 and Nanog promoters and repressive H3K9me3 and H3k27me3 at the Brachyury promoter. Additionally, the activation of signaling pathways involved in pluripotency, such as Gsk3-β/β-catenin, was observed, in addition to activation of PI3 K/Akt, which is consistent with the protection of mESCs from cell death. Finally, a decrease in cell proliferation coincides with cell cycle arrest in G2/M. Our results provide novel insights into NO-mediated gene regulation and cell proliferation and suggest that NO is necessary but not sufficient for the maintenance of pluripotency and the prevention of cell differentiation. J. Cell. Biochem. 117: 2078-2088, 2016. © 2016 Wiley Periodicals, Inc.
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|RED-TERCEL, Seville, Spain.
authors: Rafael
authors: R
authors: Tapia-Limonchi
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.
authors: Gladys M
authors: GM
authors: Cahuana
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.
authors: Estefania
authors: E
authors: Caballano-Infantes
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, Fundación Progreso y Salud, Seville, Spain.
authors: Carmen
authors: C
authors: Salguero-Aranda
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.
authors: Amparo
authors: A
authors: Beltran-Povea
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.
authors: Ana B
authors: AB
authors: Hitos
authors: RED-TERCEL, Seville, Spain.|Andalusian Center for Molecular Biology and Regenerative Medicine, Fundación Progreso y Salud, Seville, Spain.
authors: Abdelkrim
authors: A
authors: Hmadcha
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|RED-TERCEL, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.
authors: Franz
authors: F
authors: Martin
authors: RED-TERCEL, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.|Andalusian Center for Molecular Biology and Regenerative Medicine, Fundación Progreso y Salud, Seville, Spain.
authors: Bernat
authors: B
authors: Soria
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|RED-TERCEL, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.
authors: Francisco J
authors: FJ
authors: Bedoya
authors: Andalusian Center for Molecular Biology and Regenerative Medicine, University Pablo de Olavide, Seville, Spain.|RED-TERCEL, Seville, Spain.|Biomedical Research Network on Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain.
authors: Juan R
authors: JR
authors: Tejedo
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects / Q000502:physiology*; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology*; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014226:Triazenes / Q000494:pharmacology
chemical_list: C105351:1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene; D014226:Triazenes; D009569:Nitric Oxide; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt
keywords: CELL DIFFERENTIATION; EMBRYONIC STEM CELLS; GENE REGULATION; NITRIC OXIDE; PLURIPOTENCY