External factors
| External factors: |
Doxorubicin |
| Category: |
Chemical compounds |
| Experiment: |
SA-β-gal activity assay//Flow cytometry |
| Description: |
Flow cytometry：Flow cytometric examination of the cell cycle distribution of NCI-H292 cells showed a significant increase of cells with DNA content corresponding to G2/M at 80 nM of doxorubicin compared to the control and 20 nM (p<0.05), whereas the cells with DNA content corresponding to G0/G1 concurrently decreased (p<0.05). SA-β-gal activity assay：The number of β-gal-positive senescent cells was markedly increased in doxorubicin treated by comparison with untreated NCI-H292 cultures. |
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Regulatory relationship
| Target gene:
|
P53//γH2AX//P21 |
| R-EF-Target gene:
|
Upregulation//Activation//Upregulation |
| Official symbol(s): |
P53//H2AX//P21 |
| Target gene experiment: |
Western blot |
| Target gene description: |
In order to determine the effect of doxorubicin on p53 protein levels western blotting analysis was performed.Doxorubicin at of 20, 40 and 80 nM induced a significant increase of p53 protein levels to 1.7 fold compared to the untreated condition (p<0.01). By using a specific antibody directed against phosphorylated H2AX, we observed that 40 and 80 nM doxorubicin promoted a significant phosphorylation of H2AX at the Ser139 residue (γH2AX) compared to 20 nM and UT, favoring its activation. Data reported, panels C-E also demonstrated that the increase of p53 was accompanied by a significant up-elevation of p21 at 80 nM doxorubicin compared to all other conditions (p<0.001). Moreover, p21 protein levels were significantly higher at 40 nM doxorubicin compared to UT and 20 nM (p<0.001). |
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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.
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