External factors
| External factors: |
Nutlin-3 |
| Category: |
Chemical compounds |
| Experiment: |
SA-β-gal activity assay//Flow cytometry//Cell morphological analysis |
| Description: |
Striking morphologic alterations characteristic of senescent cells were indeed observed in SK-N-SH cells on nutlin-3 administration, including a flattened and enlarged cell shape with increased cytoplasmic granularity. Gene expression of the cell cycle inhibitor CDKN1A was increased 14-fold after 24 hours of incubation with 16 μmol/L nutlin-3, with ～70% of cells residing in G1 phase. Treatment with 16 μmol/L nutlin-3 significantly enhanced the number of SA-β-gal-expressing cells with rapid kinetics (411 per mm2 after 24 hours; 95% CI, 371-451; P < 0.0001), suggesting that nutlin-3 does not just select for SA-β-gal-expressing cells but Activately induces a senescence-like phenomenon in surviving SK-N-SH cells. After 7 days of exposure to 16 μmol/L nutlin-3, ～100% of SK-N-SH cells not subject to apoptotic cell death stained positive for SA-β-gal and had acquired a senescence-like morphology. |
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Regulatory relationship
| Official symbol(s): |
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TP53
|
| Pathway experiment: |
Western blot |
| Pathway description: |
Western blot analysis of p53 and p21WAF1/CIP1 expression showed induction of a p53 response after treatment with 16 μmol/L nutlin-3 for 24 hours in all three types of control-infected SK-N-SH cells . |
Aging network
Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.