| External factors: | Compound fuling granule |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Ovarian cancer |
| Experiment: | SA-β-gal activity assay//Flow cytometry//Western blot//RT-PCR//Immunofluorescence |
| Description: | The cell cycle distribution of CFG-treated HEY and SKOV3 cells, determined by flow cytometry analysis, indicated that the percentage of S + G2/M was greatly decreased in HEY cells after CFG treatment, whereas in SKOV3 cells it was actually slightly upregulated.Quantitative PCR also showed that the cell cycle associated proteins were regulated by CFG, and this phenomenon was more obvious in HEY than in SKOV3 cells. Similarly, immunofluorescence staining of HEY cells with antibodies against Cdt1 (red, G1 phase) and Geminin (green, G2 phase) revealed that proportion of cells in the G2 phase was also reduced after CFG treatment. Furthermore, the analysis of the cell cycle related proteins by Western blotting showed that the expression of these proteins was indeed significantly changed in CFG treated cells.β-gal staining showed that CFG could induce cellular senescence in HEY cells only. |
| Regulatory pathway: | Akt-GSK3β |
| R-EF-Pathway: | Downregulation |
| Official symbol(s): | AKT |
| Pathway experiment: | Western blot |
| Pathway description: | The results revealed a specific reduction in the level of pAKT protein in HEY and SKOV3 cells treated with CFG, compared with that of untreated cells, as well as with that of cells singly treated with TGFβ1, as controls. The total AKT level, however, remained unaffected by all treated conditions. The expression levels of AKT downstream substrates Bcl-Xl, BAD, GSK3β, SNAIL, and SLUG were also assessed. CFG treatment also decreased the Bcl-Xl/BAD complex, pGSK3β and SLUG without affecting the nonphosphorylated form of GSK3. |
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