| Gene name: | NQO1 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | 2BS |
| Experiment: | SA-β-gal activity assay//qRT-PCR//Cell proliferation assay//BrdU assay |
| Description: | NQO1 depletion resulted in continuous cell growth compared with corresponding control lentiviral vector (Cont) infected cells . In addition, the morphology changed during the OIS, with senescent cells exhibiting the typical enlarged and flattened shape. Five days after introducing RasG12V, NQO1 depleted 2BS cells showed a different morphology compared with control 2BS cells. Accordingly, we found that 2BS cells with NQO1 depletion reduced with SA-β-gal staining after expressing RasG12V. We observed that knockdown of NQO1 resulted in an increased percentage of 2BS cells with BrdU incorporation after expressing RasG12V. This result suggests that NQO1 depletion delayed the onset of RasG12V-induced cellular senescence. |
| Target gene: | P53 |
| Official symbol(s): | P53 |
| R-AG-Target gene: | Upregulation |
| Subcategory: | Phosphorylation |
| Target gene experiment: | Western blot |
| Target gene description: | We found that NQO1 depletion clearly impeded p53 accumulation during OIS. |
| Regulatory pathway: | NRF2-KEAP1 |
| R-AG-Pathway: | -- |
| Official symbol(s): | NFE2L2-KEAP1 |
| Pathway experiment: | Western blot//CHIP//qPCR//Knockdown//Luciferase reporter assay |
| Pathway description: | MAF and KEAP1 were immunoprecipitated from 2BS cells, and western blot analysis verified that during OIS NRF2 was dislocated from KEAP1 and bound with MAF, suggesting that OIS activates NRF2 and results in NRF2-mediated up-regulation of NQO1. Next, we detected whether NRF2 bound to the endogenous NQO1 promoter by chromatin immunoprecipitation (ChIP) assay. Interaction between NRF2 and the NQO1 promoter region was detected by qPCR. As expected, interaction between NRF2 and NQO1 promoter only observed in senescent 2BS. We found that knockdown of NRF2 diminished its interaction with the NQO1 promoter. In addition, we also determined whether NRF2 affected NQO1 promoter activity with ARE by the luciferase reporter assay. |
Annotation:
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