| External factors: | Adriamycin |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Lung cancer |
| 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. |
| Target gene: | BMP4 |
| R-EF-Target gene: | Upregulation |
| Official symbol(s): | BMP4 |
| 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 |
| R-EF-Pathway: | Activation |
| Official symbol(s): | 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. |
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