| Gene name: | TGFB1 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | HCEC |
| Experiment: | SA-β-gal activity assay//Flow cytometry//qPCR//Western blot |
| Description: | Cell cycle analysis by flow cytometer showed that the HCECs accumulated at G1 phase with a concomitant depletion of S phase cells after TGF-β1 exposure, suggesting that cell cycle arrest during HCECs senescence induced by TGF-β1 occurred at G1 phase, while H2O2 induced an obvious G2/M phase arrest. In association with the G1 arrest, we also found TGF-β1 increased the percentage of SA-β-gal staining cells and concomitantly increased the expression of p16 and p21, as analyzed by real-time PCR or western blot. In order to further confirm the effect of TGF-β on cellular senescence, we interrupted the TGF-β signaling pathway using a specific inhibitor, LY364947. When treated with LY364947, the percentage of SA-β-gal staining cells was significantly decreased, and the levels of p21 and p16 were also downregulated. |
| Regulatory pathway: | NF-κB |
| R-AG-Pathway: | Activation |
| Official symbol(s): | NFKB1 |
| Pathway experiment: | Western blot |
| Pathway description: | Compared with pre-senescent cells, TGF-β1 treatment led to significant reduction in IκBα protein levels. IκBα is a canonical and specific inhibitor of NF-κB, and its reduction suggested elevated NF-κB activity in TGFβ-induced senescent cells; our subsequent results confirmed this. IκBα loss is the key step that immediately releases cytoplasmic NF-κB for nuclear translocation and the subsequent phosphorylation of p65, a subunit of NF-κB. Therefore, we measured the phosphorylation of p65 in HCECs treated with or without TGF-β. Much more phosphorylation of p65was found in TGF-β-treated HCECs than in untreated cells. |
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