| Gene name: | DUSP16 |
| Aging type: | Prevent |
| Aging characteristic: | Others |
| Tissue type: | Human HCC sample,Tumor tissue |
| Cell name: | PLC-PRF-5 |
| Experiment: | Knockdown//SA-β-gal activity assay//TUNEL assay//Flow cytometry//BrdU assay |
| Description: | TUNEL and Propidium Iodide (PI) staining, respectively. TUNEL-positive signals could be detected in DNase I or chemotherapeutic drug doxorubicin treated cells, but not in DUSP16-silenced cells ,By contrast, we found that upon DUSP16 knockdown, the PLC/PRF/5 cells were blocked in the G1 phase, suggesting that these cells were unable to initiate DNA replication and maintained a growth-arrested state. We also noticed that DUSP16-silenced cells exhibited a lower BrdU incorporation rate when compared with control cells.In accordance with the impaired cell cycle transition, the cell cycle-related proteins CDK1, CDK2, CDK4 and cyclin E were markedly down-regulated in DUSP16-silenced cells. SA-β-Gal activity is a widely used marker for measuring cellular senescence. Intriguingly, the percentage of SA-β-Gal-positive cells significantly increased upon DUSP16 silencing. |
| Regulatory pathway: | P53//RB |
| R-AG-Pathway: | --//-- |
| Official symbol(s): | TP53//retinoblastoma |
| Pathway experiment: | Knockdown//SA-β-gal activity assay//Western blot//qRT-PCR |
| Pathway description: | Upon DUSP16 knockdown, p53 downstream effectors were dramatically up-regulated .In addition, we found that upon DUSP16 knockdown, the increase of SA-β-Gal-positive cells and the decrease of the number of viable cells were efficiently attenuated by shRNA-mediated p53 silencing .Interestingly, in PLC/PRF/5 cells, we identified that decreased levels of CDKs upon DUSP16 silencing led to reduced phosphorylation of Rb , suggesting that the activation of Rb is involved in DUSP16 silencing-induced senescence in p53 mutant liver cancer cells .In addition, in the HepG2 cell line that carries wild-type p53, our results showed increased phosphorylation of p53, as well as, reduced phosphorylation of Rb upon DUSP16 silencing ,suggesting that both p53 and Rb are likely downstream effectors of DUSP16 silencing-induced senescence in p53 wild-type liver cancer cells. |
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