Regulation of cardiotrophin-1 expression during mouse embryonic stem cell differentiation by hypoxia and reactive oxygen species
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Cardiomyogenesis in differentiating mouse embryonic stem (ES) cells is promoted by cardiotrophin-1 (CT-1), a member of the IL-6 interleukin superfamily and acts through the gp130 cytokine receptor. It was shown that prooxidants (menadione, hydrogen peroxide) as well as chemical (CoCl2) and physiological (1% O2) hypoxia increased CT-1 as well as HIF-1a protein and mRNA expression in embryoid bodies, indicating that CT-1 expression is regulated by reactive oxygen species (ROS) and hypoxia. Treatment with either prooxidants or chemical hypoxia increased gp130phosphorylation and protein expression of NADPH-oxidase subunits p22-phox, p47-phox, p67-phox, as well as Nox-1 and Nox-4 mRNA. Consequently, inhibition of NADPH-oxidase activity by diphenylen iodonium chloride (DPI) and apocynin abolished prooxidant- and chemical hypoxia-induced up-regulation of CT-1 and HIF-1. Prooxidants and chemical hypoxia activated ERK1,2, JNK and p38 as well as PI3-kinase, JAK2 and STAT3. The pro-oxidant and CoCl2-mediated up-regulation of CT-1 and HIF-1 was significantly inhibited in the presence of the ERK1,2 antagonist UO126, the JNK antagonist SP600125, the p38 antagonist SKF86002, the PI3-kinase antagonist LY294002, the JAK-2 antagonist AG490 as well as in the presenceof free radical scavengers. Moreover, developing embryoid bodies derived from HIF-1a-/--ES cells lack cardiomyogenesis, and prooxidants as well as chemical hypoxia failed to upregulate CT-1 expression. Treatment of cells obtained by collagenase dissociation of beating EBs with pro-oxidants and chemical hypoxia resulted in the translocation of CT-1 into the nucleus within 2-4 h. In summary our data demonstrate that CT-1 expression in ES cells is regulated by ROS and HIF-1 and imply a crucial role of CT-1 in survival and proliferation of ES cell-derived cardiac cells. The importance of CT-1 nuclear transport to CT-1 mediated gene expression is still to be determined.Verknüpfung zu Publikationen oder weiteren Datensätzen
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Journal of Cell Science, 119 (2006), S. 1043-1052