External factors
| External factors: |
Adriamycin |
| Category: |
Chemical compounds |
| Experiment: |
MTT assay//SA-β-gal activity assay//Cell morphological analysis |
| Description: |
MTT assays showed that adriamycin was able to decrease the proliferation indices of NCI-H460 and A549 cells at 500 nM and 2 M, respectively. We then found that NCIH460 and A549 cells treated with adriamycin exhibited phenotypic changes that resembled those observed in cells undergoing senescence,including theintensified SA-β- gal staining, flattened cell morphology, and enlarged cell size, in just 1 week.We next examined the endogenous SA-β-gal (pH 6.0) activity.A great increase in SA-β-gal activity was seen 6 days after adriamycin treatment,in comparison with untreated cells. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation |
| Target gene experiment: |
SA-β-gal activity assay//Knockdown//Western blot//qRT-PCR |
| Target gene description: |
We then showed that suppression of endogenous BMP4 expression restrained the adriamycin-induced senescence ,eliciting the critical role of BMP4 in thisprocess. Representative results show that induction of the BMP4 expression by DOX was both doseand time-dependent. Moreover, Western blots revealed that the induction of BMP4 expression by DOX was well equivalent to the extent of BMP4 up-regulation by adriamycin.RT-PCR and quantitative real-time PCR data demonstrated that the BMP4 mRNA level was greatly up-regulated when NCI-H460 and A549 cells were treated with adriamycin at 500 nM and 2 M, respectively. |
| Regulatory pathway:
|
BMP-Smad |
| Official symbol(s): |
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BMPR2-SMAD4
|
| Pathway experiment: |
SA-β-gal activity assay//Knockdown//Western blot |
| Pathway description: |
The ectopic expression of Smad6- myc was first confirmed by Western blot analysis . We then showed that suppression of the BMP-Smad pathway by Smad6 did interfere with the BMP4-induced premature senescence, as revealed by SA-β-gal activity assay, SA-β-gal staining, and cellular immunofluorescence. |
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.