Dynamic monitoring of apoptosis in chemotherapies with multiple fluorescence reporters.
Abstract
The aim of the study is to dynamically and non-invasively monitor the temporal relationship among caspase-3, BID, and cytochrome c in chemotherapy. 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. 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. 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.
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Apoptosis
Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis