Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques

Huiying Wang; Tongsheng Chen; Lei Sun

doi:10.1117/12.761440

18 February 2008 Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques

Huiying Wang, Tongsheng Chen, Lei Sun

Author Affiliations +

Proceedings Volume 6857, Biophotonics and Immune Responses III; 68570J (2008) https://doi.org/10.1117/12.761440
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

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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Huiying Wang, Tongsheng Chen, and Lei Sun "Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques", Proc. SPIE 6857, Biophotonics and Immune Responses III, 68570J (18 February 2008); https://doi.org/10.1117/12.761440

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