The stabilization of hypoxia inducible factor modulates differentiation status and inhibits the proliferation of mouse embryonic stem cells
Language English Country Ireland Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
26723917
DOI
10.1016/j.cbi.2015.12.007
PII: S0009-2797(15)30142-3
Knihovny.cz E-resources
- Keywords
- DMOG, Embryonic stem cells, HIF-1, Hypoxia, JNJ-42041935, Prolyl hydroxylase,
- MeSH
- Amino Acids, Dicarboxylic chemistry pharmacology MeSH
- Benzimidazoles chemistry pharmacology MeSH
- Cell Differentiation drug effects MeSH
- Cell Cycle drug effects MeSH
- Hypoxia-Inducible Factor 1 metabolism MeSH
- Hypoxia metabolism MeSH
- Enzyme Inhibitors chemistry pharmacology MeSH
- Cells, Cultured MeSH
- Mouse Embryonic Stem Cells cytology drug effects metabolism MeSH
- Mice MeSH
- Cell Proliferation drug effects MeSH
- Hypoxia-Inducible Factor-Proline Dioxygenases antagonists & inhibitors metabolism MeSH
- Pyrazoles chemistry pharmacology MeSH
- Protein Stability drug effects MeSH
- Cell Survival drug effects MeSH
- Dose-Response Relationship, Drug MeSH
- Structure-Activity Relationship MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 1-(5-chloro-6-(trifluoromethoxy)-1H-benzoimidazol-2-yl)-1H-pyrazole-4-carboxylic acid MeSH Browser
- Amino Acids, Dicarboxylic MeSH
- Benzimidazoles MeSH
- Hypoxia-Inducible Factor 1 MeSH
- Enzyme Inhibitors MeSH
- oxalylglycine MeSH Browser
- Hypoxia-Inducible Factor-Proline Dioxygenases MeSH
- Pyrazoles MeSH
Hypoxic conditions are suggested to affect the differentiation status of stem cells (SC), including embryonic stem cells (ESC). Hypoxia inducible factor (HIF) is one of the main intracellular molecules responsible for the cellular response to hypoxia. Hypoxia stabilizes HIF by inhibiting the activity of HIF prolyl-hydroxylases (PHD), which are responsible for targeting HIF-alpha subunits for proteosomal degradation. To address the impact of HIF stabilization on the maintenance of the stemness signature of mouse ESC (mESC), we tested the influence of the inhibition of PHDs and hypoxia (1% O2 and 5% O2) on spontaneous ESC differentiation triggered by leukemia inhibitory factor withdrawal for 24 and 48 h. The widely used panhydroxylase inhibitor dimethyloxaloylglycine (DMOG) and PHD inhibitor JNJ-42041935 (JNJ) with suggested higher specificity towards PHDs were employed. Both inhibitors and both levels of hypoxia significantly increased HIF-1alpha and HIF-2alpha protein levels and HIF transcriptional activity in spontaneously differentiating mESC. This was accompanied by significant downregulation of cell proliferation manifested by the complete inhibition of DNA synthesis and partial arrest in the S phase after 48 h. Further, HIF stabilization enhanced downregulation of the expressions of some pluripotency markers (OCT-4, NANOG, ZFP-42, TNAP) in spontaneously differentiating mESC. However, at the same time, there was also a significant decrease in the expression of some genes selected as markers of cell differentiation (e.g. SOX1, BRACH T, ELF5). In conclusion, the short term stabilization of HIF mediated by the PHD inhibitors JNJ and DMOG and hypoxia did not prevent the spontaneous loss of pluripotency markers in mESC. However, it significantly downregulated the proliferation of these cells.
References provided by Crossref.org
Hypoxia favors myosin heavy chain beta gene expression in an Hif-1alpha-dependent manner
