External factors
| Category: |
Chemical compounds |
| Experiment: |
MTT assay// FACS analysis//Immunofluorescence//Western blot//SA-β-gal activity assay |
| Description: |
RSV inhibited the proliferation of U2OS cells in a dose-dependent manner. Analysis of cell cycle distribution by flow cytometry indicated that RSV dose-dependently induced S-phase arrest. There was a remarkable increase in the amount of γ-H2AX, a marker for DNA double-strand breaks, in RSV-treated cells, as revealed by immunofluorescence. Meanwhile, ATM, a master regulator of DNA damage response, was activated by RSV.In addition, typically of cells experiencing replication stress, the level of phosphorylated CHK1 level was increased. Consistent with previous report in HCT116 colon cancer cells12, there was a significant induction of cellular senescence. It appeared that the cells arrested at S-phase could progress to senescence, as the majority of cells remained in S-phase when they became senescent. |
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Regulatory relationship
| Target gene:
|
NUCLEOSIDES//P53//CXCR2 |
| R-EF-Target gene:
|
--//--//-- |
| Official symbol(s): |
NUCLEOSIDES//P53//CXCR2 |
| Target gene experiment: |
RT-PCR//Flow cytometry//SA-β-gal activity assay |
| Target gene description: |
Importantly, RSV-induced senescence was greatly reduced by nucleosides. We tested the inhibitory effect of nucleosides on RSV-induced senescence in two additional cell lines HT1080 (fibrosarcoma) and A549 (lung cancer) and obtained similar results. Indeed, CXCR2 transcription was significantly increased by RSV in a dose- and time-dependent manner . The increased expression of CXCR2 was confirmed by flow cytometry analysis of CXCR2 (CD182). However, after CXCR2 expression reached its peak level at day 5, it began to subside in the following days.We confirmed that the upregulation of CXCR2 following RSV treatment was dependent on p53, since the expression of CXCR2, like that of p21, in response to RSV was greatly reduced when p53 was depleted. Importantly, depletion of CXCR2 greatly attenuated RSV-induced senescence. |
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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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