Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques
2008
Caspase-3 is a key activated death protease, which catalyzes the specific cleavage of many cellular proteins and induces
DNA cleavage eventually. In this report, cells were treated with mitomycin C (MMC) at different concentration and its
activity was detected by cell counting kit (CCK-8). Based on results of CCK-8, cells were treated with 10μg/mL MMC
and Hoechst 33258 has been used to observe cell apoptosis. Fluorescence resonance energy transfer (FRET) and
confocal microscopy have been used to the effect of MMC on the caspase3 activation in living cells. Human lung
adenocarcinoma cells (ASTC-a-1) was transfected with plasmid SCAT3 (pSCAT3)/CKAR FRET receptor. Acceptor
photobleaching techniques of FRET plasmid has been used to destruct fluorophore of cells stably expressing SCAT3
reporter on a fluorescence confocal microscope. The activity of caspase3 can be analyzed by FRET dynamics of SCAT3
in living cells. Our results show that MM C can induce ASTC-a-1 cell apoptosis through activation of caspase3.
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