| Gene name: | BAG2 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | TRE293 |
| Experiment: | Western blot//BrdU assay//SA-β-gal activity assay//SAHF |
| Description: | To assess the functional role of BAG2 in senescence, we generated a TRE293 cell line with inducible BAG2 (TRE293-BAG2) in which the BAG2 expression is triggered by the tetracycline regulatory (Tet-on) system. Like other senescent cells, the TRE293-BAG2 cells exhibited cell cycle arrest and slow proliferation and a significant increase in SA-β-Gal staining. In a senescence cell, it is basically accepted that the abundant enhancement of senescence-associated heterochromatic foci (SAHF) with heterochromatin proteins such as HP1 represses expression of proliferation-promoting genes. The senescence induced by BAG2 was further confirmed by examining changes in the chromatin structure. |
| Regulatory pathway: | P21 |
| R-AG-Pathway: | Activation |
| Official symbol(s): | CDKN1A |
| Pathway experiment: | Western blot//Knockdown//SA-β-gal activity assay |
| Pathway description: | We monitored the levels of typical biomarkers of senescence in TRE293-BAG2 cells, (p21CIP1, p16INK4a, or p27KIP1). Western blotting revealed no significant change in the levels of p16INK4a or p27KIP1. However, there was a strong accumulation of p21CIP1, suggesting that the senescence induced by BAG2 was likely via the p21CIP1 pathway, which was also observed in H520-BAG2 cells. This interpretation was further supported by BAG2 knock-down experiments.The level of p21CIP1 protein was reduced in the TRE293 cells treated with BAG2 small interfering RNA (siRNA). To determine if the senescence induced by BAG2 was mediated only through the p21CIP1 pathway, we transfected p21CIP1 shRNA into TRE293- BAG2 cells. We found that senescence was almost fully restored when p21CIP1 was down-regulated. |
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