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Imaging of activated caspase-3 in living cell by fluorescence resonance energy transfer during photosensitization-induced apoptosis
Author(s): Yunxia Wu; Da Xing; Qun Chen; Yonghong Tang
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Paper Abstract

Photodynamic therapy (PDT) is a novel and promising cancer treatment that employs a combination of a photosensitizing chemical and visible light, induces apoptosis in cell, and activation of caspase-3 is considered to be the final step in many apoptosis pathways. The changes of caspase-3 activation in cell during TNFα- and photodynamic therapy-induced apoptosis was measured by fluorescence resonance energy transfer (FRET) analysis. FRET probe consisting of fusions of an enhanced cyan fluorescent protein (ECFP), Venus and a linker peptide containing the caspase-3 cleavage sequence DEVD was utilized. Therefore, activated caspase-3 cleaved the linker peptide of FRET probe and disrupted the FRET signal. Human lung adenocarcinoma cell line (ASTC-a-1) were stably transfected with the plasmid (ECFP-DEVD-Venus) and then were treated by TNF-α and PDT, respectively. Experimental results indicated that caspase-3 activation resulted in cleavage of linker peptide and subsequent disruption of the FRET signal during TNFα- and photodynamic therapy-induced apoptosis, and that the activation of caspase-3 induced by photodynamic therapy was faster than that induce by TNF-α. The study supports that using FRET technique and different recombinant substrates as FRET probes could be used to detect the process of PDT-induced apoptosis and provide a new means to investigate apoptotic mechanism of PDT.

Paper Details

Date Published: 18 January 2005
PDF: 8 pages
Proc. SPIE 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II, (18 January 2005); doi: 10.1117/12.572773
Show Author Affiliations
Yunxia Wu, South China Normal Univ. (China)
Da Xing, South China Normal Univ. (China)
Qun Chen, South China Normal Univ. (China)
Yonghong Tang, South China Normal Univ. (China)

Published in SPIE Proceedings Vol. 5630:
Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II
Britton Chance; Mingzhe Chen; Arthur E. T. Chiou; Qingming Luo, Editor(s)

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