| Gene name: | MIR17 |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | Heart |
| Cell name: | MCF |
| Experiment: | SA-β-gal activity assay//Cell apoptosis assay//qRT-PCR |
| Description: | Transfection of the miR-17-3p inhibitor enhanced MCF senescence and apoptosis in cells treated with H2O2, whereas expression of miR-17-3p repressed senescence and apoptosis.Consistent with these results, we found that the aging hearts expressed significantly lower levels of miR-17-3p compared with the levels in hearts from young animals. |
| Target gene: | PAR4 |
| Official symbol(s): | PAR4 |
| R-AG-Target gene: | Downregulation |
| Subcategory: | Unclear |
| Target gene experiment: | Luciferase reporter assay//Western blot//SA-β-gal activity assay |
| Target gene description: | Luciferase activity was repressed when the construct (Luc-Par4) was co-transfected with miR-17-3p mimic, and the repression was abolished when the target site was mutated, thus confirming direct targeting of Par4 by miR-17-3p. Protein lysates prepared from miR-17- and vector-transfected cells were subjected to western blotting, which showed that ectopic transfection of miR-17 repressed Par4 expression significantly. Ectopic expression of Par4 promoted senescence when the cells were cultured in serum-free medium or treated with H2O2. |
| Regulatory pathway: | CEBPB-FAK |
| R-AG-Pathway: | -- |
| Official symbol(s): | CEBPB-PTK2 |
| Pathway experiment: | Western blot//IP//RT-PCR//PCR//ChIP |
| Pathway description: | The ChIP assay indicated that Par4 might bind to the CEBPB promoter directly and repress CEBPB transcription, whereas CEBPB could bind to the FAK promoter and enhance FAK transcription. By using western blotting, we confirmed that silencing Par4 promoted the expression of CEBPB and FAK.The Par4-expressing cells showed high levels of Par4 and low levels of CEBPB and FAK, and the regulation appeared to occur at the transcriptional level. In addition, ectopic transfection of CEBPB enhanced FAK expression at the mRNA and protein levels. By ChIP, immunoprecipitated exogenous Par4 was shown to pull down more CEBPB promoter DNA, whereas immunoprecipitated exogenous CEBPB could pull down more FAK promoter DNA. |
Annotation:
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