External factors: | Cyclosporine A |
Aging type: | Accelerate |
Aging characteristic: |
Category: | Chemical compounds |
Phenotype: | Glioma |
Experiment: | SA-β-gal activity assay//Flow cytometry//Western blot//Cell morphological analysis |
Description: | CsA increases the number of cells in the G1 phase of cell cycle and significantly reduces the number of proliferating cells in the S phase. p21WAF1/Cip1 protein is an universal inhibitor of cyclin kinases and plays an important role in inhibiting cell proliferation. Western blot analysis of p21WAF1/Cip1 level was performed on human glioma cells: U87-MG (wild-type p53), T98G and U373-MG (p53 mutated) following CsA treatment.The levels of p21WAF1/Cip1 protein increased in all examined glioma cell lines. The highest up-regulation was observed in T98G and the moderate p21WAF1/Cip1 protein accumulation was observed in U373-MG and U87MG cells. In T98G cells the increase of p21WAF1/Cip1 protein was observed at 5 h, reaching the high level at 15 h, which remained elevated 25 h after CsA treatment.Untreated U87-MG and T98G cells demonstrated very rare cells positive for the SA-β-Gal staining. After 24 h of CsA treatment the percentage of cells positive for SA-β-Gal was doubled in U87-MG cells compared to control, while only few T98G cells were positively stained. SA-β-Gal positive U87-MG cells increased in size and flattened out, thereby attaining a morphology of senescent-like cells. |
Regulatory pathway: | -- |
R-EF-Pathway: | -- |
Pathway experiment: | -- |
Pathway description: | -- |
Annotation: