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This study investigated the signal molecules linking the alteration in 2-dexoyglucose (2-DG) uptake and DNA synthesis in mouse embryonic stem (ES) cells under hypoxia. Hypoxia increased the 2-DG uptake and GLUT-1 protein expression level while the undifferentiated state of ES cells and cell viability were not affected by the hypoxia (1 - 48h). Subsequently, [(3)H] thymidine incorporation was significantly increased at 12 hours of hypoxic exposure. Hypoxia increased the Ca(2+) uptake and PKC beta (I), epsilon, and zeta translocation from the cytosol to the membrane fraction. Moreover, hypoxia increased the level of p44/42 mitogen-activated protein kinases (MAPKs) phosphorylation and hypoxia inducible factor-1alpha (HIF-1alpha) in a time-dependent manner. On the other hand, inhibition of these pathways blocked the hypoxia-induced increase in the 2-DG uptake and GLUT-1 protein expression level. Under hypoxia, cell cycle regulatory protein expression [cyclin D1, cyclin E, cyclin-dependent kinase (CDK) 2, and CDK 4] were increased in a time-dependent manner, which were blocked by PD 98059. pRB protein was also increased in a time-dependent manner. In conclusion, under hypoxia, there might be a parallel relationship between the expression of GLUT1 and DNA synthesis, which is mediated by the Ca(2+) /PKC, MAPK, and the HIF-1alpha signal pathways in mouse ES cells.

cellular physiology and biochemistry

Effect of Hypoxia on 2-Deoxyglucose Uptake and Cell Cycle Regulatory Protein Expression of Mouse Embryonic Stem Cells: Involvement of Ca&lt;sup&gt;2+&lt;/sup&gt; /PKC, MAPKs and HIF-1α