Dynamic monitoring of apoptosis in chemotherapies with multiple fluorescence reporters.
OBJECTIVE
The aim of the study is to dynamically and non-invasively monitor the temporal relationship among caspase-3, BID, and cytochrome c in chemotherapy.


METHODS
ASTC-a-1 cells expressing the corresponding fluorescence reporters were treated with Taxol or cisplatin and imaged using FRET and fluorescence overlapping technique. Western blot was performed to validate the fluorescence analysis.


RESULTS
In fluorescence imaging analysis, Taxol-induced apoptosis showed caspase-3 activation (13 h 50 min) was prior to BID cleavage (15 h 10 min) and subsequent significant cytochrome c release (17-18 h 20 min), whereas the cisplatin-induced apoptosis showed BID cleavage (5 h 40 min) and significant cytochrome c release (7-8 h 20 min) were prior to caspase-3 activation (14 h 20 min). Western blot further validated the results above.


CONCLUSIONS
The new approach successfully reveals the difference in temporal signaling apoptosis events between Taxol and cisplatin. It may help us come to a better understanding of the detailed mechanisms in chemotherapeutic-agents-induced apoptosis.
Li
Yunlong
Y
MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou, 510631, China.
Xing
Da
D
Chen
Qun
Q
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D045304:Cytochromes c / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D017239:Paclitaxel / Q000494:pharmacology
D000970:Antineoplastic Agents; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D049452:Green Fluorescent Proteins; D045304:Cytochromes c; D053148:Caspase 3; D017239:Paclitaxel; D002945:Cisplatin