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Journal of Biomedical Optics

Activation of caspase-3 noninvolved in the bystander effect of the herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) system
Author(s): Zhihong Zhang; Juqiang Lin; Jun Chu; Yan Ma; Shaoqun Zeng; Qingming Luo
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Paper Abstract

Use of the herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) system is one of the promising approaches in the rapidly growing area of gene therapy. The "bystander effect," a phenomenon in which HSV-tk+ cells exposed to GCV are toxic to adjacent HSV-tk-; cells, was reported to play an important role in suicide gene therapy. However, the mechanism by which HSV-tk/GCV induces the bystander effect is poorly understood. We monitored the activation of caspase-3 in living cells induced by the HSV-tk/GCV system using a genetically encoded fluorescence resonance energy transfer (FRET) probe CD3, a caspase-3 recognition site fused with a cyan fluorescent protien (CFP) and a red fluorescent protein (DsRed) which we reported and named in a previous paper. Fluorescence protein (FP)–based multicolor cellular labeling, combined with the multichannel fluorescence imaging and FRET imaging techniques, provides a novel and improved approach to directly determine whether the activation of caspase-3 involved in the HSV-tk/GCV system induces cell apoptosis in tk gene-expressing cells and their neighboring cells. FRET ratio images of CD3, and fluorescence images of the fusion protein of thymidine kinase linked with green fluorescent protein (TK-GFP), indicated that HSV-tk/GCV system–induced apoptosis in human adenoid cystic carcinoma (ACC-M) cells was via a caspase-3 pathway, and the activation of caspase-3 was not involved in the bystander effect of HSV-tk/GCV system.

Paper Details

Date Published: 1 May 2008
PDF: 5 pages
J. Biomed. Opt. 13(3) 031209 doi: 10.1117/1.2937830
Published in: Journal of Biomedical Optics Volume 13, Issue 3
Show Author Affiliations
Zhihong Zhang, Huazhong Univ. of Science and Technology (China)
Juqiang Lin, Huazhong Univ. of Science and Technology (China)
Jun Chu, Huazhong Univ. of Science and Technology (China)
Yan Ma, Huazhong Univ. of Science and Technology (China)
Shaoqun Zeng, Huazhong Univ. of Science and Technology (China)
Qingming Luo, Huazhong Univ. of Science and Technology (China)


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