Monitoring apoptosis of TK-GFP-expressing ACC-M cells induced by ACV using FRET technique

Tao Xiong; Zhihong Zhang; Juqiang Lin; Jie Yang; Shaoqun Zeng; Qingming Luo

doi:10.1117/12.662591

19 May 2006 Monitoring apoptosis of TK-GFP-expressing ACC-M cells induced by ACV using FRET technique

Tao Xiong, Zhihong Zhang, Juqiang Lin, Jie Yang, Shaoqun Zeng, Qingming Luo

Proceedings Volume 6218, Chemical and Biological Sensing VII; 62180L (2006) https://doi.org/10.1117/12.662591
Event: Defense and Security Symposium, 2006, Orlando (Kissimmee), Florida, United States

Abstract

Apoptosis is an evolutionary conserved cellular process that plays an important role during development, but it is also involved in tissue homeostasis and in many diseases. To study the characteristics of suicide gene system of the herpes simplex virus thymidine kinase (HSV-tk) gene in tumor cells and explore the apoptosis phenomena in this system and its effect on the human adenoid cystic carcinoma line ACC-M cell, we detected apoptosis of CD3- (ECFP-CRS-DsRed) and TK-GFP-expressing ACC-M (ACC-M-TK-GFP-CD3) cells induced by acyclovir (ACV) using fluorescence resonance energy transfer (FRET) technique. CD3 is a FRET-based indicator for activity of caspase-3, which is composed of an enhanced cyan fluorescent protein, a caspase-3 sensitive linker, and a red fluorescent protein from Discosoma with efficient maturation property. FRET from ECFP to DsRed could be detected in normal ACC-M-TK-GFP-CD3 cells, and the FRET efficient was remarkably decreased and then disappeared during the cells apoptosis induced by ACV. It was due to the activated caspase-3 cleaved the CD3 fusion protein. In this study, the results suggested that the ACV-induced apoptosis of ACC-M-TK-GFP-CD3 cells was through caspase-3 pathway.

Citation Download Citation

Tao Xiong, Zhihong Zhang, Juqiang Lin, Jie Yang, Shaoqun Zeng, and Qingming Luo "Monitoring apoptosis of TK-GFP-expressing ACC-M cells induced by ACV using FRET technique", Proc. SPIE 6218, Chemical and Biological Sensing VII, 62180L (19 May 2006); https://doi.org/10.1117/12.662591

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KEYWORDS

Fluorescence resonance energy transfer

Cell death

Tumors

Green fluorescent protein

Fluorescent proteins

Luminescence

Microscopy

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