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Proceedings Paper

Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques
Author(s): Huiying Wang; Tongsheng Chen; Lei Sun
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Paper 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.

Paper Details

Date Published: 18 February 2008
PDF: 7 pages
Proc. SPIE 6857, Biophotonics and Immune Responses III, 68570J (18 February 2008); doi: 10.1117/12.761440
Show Author Affiliations
Huiying Wang, South China Normal Univ. (China)
Tongsheng Chen, South China Normal Univ. (China)
Lei Sun, South China Normal Univ. (China)

Published in SPIE Proceedings Vol. 6857:
Biophotonics and Immune Responses III
Wei R. Chen, Editor(s)

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