External factors: | ABT-737 |
Aging type: | Accelerate |
Aging characteristic: |
Category: | Chemical compounds |
Phenotype: | Prostate cancer |
Experiment: | MTT assay//BrdU assay//Colony formation assay |
Description: | MTT assay: Long-term exposure of PV-10 cells to ABT-737 induced significant growth inhibition and a G1/S cell cycle blockade. Brdu assay//Colony formation assay: a dose-dependent inhibition of colony formation and an inhibition of BrdU incorporation was seen after the addition of ABT-737 to both PV-10 and 22Rv1 cells. |
Target gene: | AMT |
R-EF-Target gene: | Activation |
Official symbol(s): | AMT |
Target gene experiment: | Western blot//SA-β-gal activity assay |
Target gene description: | Western blot: In a pool of PV-10 cells ATM levels were decreased by shRNA, we found that a marked reduction in ATM levels prevented ABT-737 from inducing γ-H2AX.SA-β-gal activity assay: ABT-737 treatment of cells with lower levels of ATM protein kinase caused a markedly reduced level of SA-β-Gal and senescence-like morphologic changes .we measured levels of IL-6 and IL-8 mRNA in PV-10-shRNAmir-ATM and PV-10-shRNAmir-control cells. QT-PCR analysis showed that ATM knockdown prevented the ABT-737–mediated induction of IL-6 and IL-8 mRNA. |
Regulatory pathway: | p53-p21 |
R-EF-Pathway: | Activation |
Official symbol(s): | TP53-CDKN1A |
Pathway experiment: | Western blot//SA-β-gal activity assay |
Pathway description: | Western blot: Both PV-10 and 22Rv1 cells when treated with ABT-737 show increased levels of both wild-type p53 and p21 protein levels. We find that ABT-737 treatment inhibits Cdk2 activity in a dose-dependent fashion in both PV-10 and 22Rv1 cells. Western blot//SA-β-gal activity assay: Western blot analysis confirmed that after ABT-737 treatment overexpression of DN p53 caused a substantial reduction in p21 and inhibited the ability of ABT-737 to induce SA-β-Gal. |
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