Real-time single cell analysis of bid cleavage and translocation in cisplatin-induced apoptosis

Lei Liu D.D.S.; Da Xing; Yihui Pei; Wei R. Chen

doi:10.1117/12.699634

13 February 2007 Real-time single cell analysis of Bid cleavage and translocation in cisplatin-induced apoptosis

Lei Liu D.D.S., Da Xing, Yihui Pei, Wei R. Chen

Proceedings Volume 6438, Biophotonics and Immune Responses II; 64380G (2007) https://doi.org/10.1117/12.699634
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

Cancer cell apoptosis can be induced by cisplatin, an efficient anticancer agent. However, its mechanism is not fully understood. Bcl-2 homology domain (BH) 3-only proteins couple stress signals to mitochondrial apoptotic pathways. Calpain-mediated cleavage of the BH3-only protein Bid into a 14 kD truncated protein (tBid) has been implicated in cisplatin-induced apoptotic pathway. We utilized a recombinant fluorescence resonance energy transfer (FRET) Bid probe to determine the kinetics of Bid cleavage during cisplatin-induced apoptosis in ASTC-a-1 cells. The cells were also co-transfected with Bid-CFP and DsRed-Mit to dynamically detect tBid translocation. Cells showed a cleavage of the Bid-FRET probe occurring at about 4-5 h after treated with 20 µM cisplatin. Cleavage of the Bid-FRET probe coincided with a translocation of tBid from the cytosolic to the mitochondria, and the translocation lasted about 1.5 h. Using real-time single-cell analysis, we first observed the kinetics of Bid cleavage and translocation to mitochondria in living cells during cisplatin-induced apoptosis.

Citation Download Citation

Lei Liu D.D.S., Da Xing, Yihui Pei, and Wei R. Chen "Real-time single cell analysis of Bid cleavage and translocation in cisplatin-induced apoptosis", Proc. SPIE 6438, Biophotonics and Immune Responses II, 64380G (13 February 2007); https://doi.org/10.1117/12.699634

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