Type

Journal Article

Authors

Jochen H M Prehn
Donat Kögel
Markus Rehm
Caoimhín G Concannon
Frank E Murray
Mary D Cannon
Heiko Düssmann
Robert O'Byrne
Barbara Köhler
Sergio Anguissola

Subjects

Pharmacology

Topics
apoptosis hct116 cells drug therapy organoplatinum compounds antagonists inhibitors hela cells oxaliplatin pathology bh3 interacting domain death agonist protein bid protein human metabolism calpain uterine cervical neoplasms humans physiology pharmacology female therapeutic use drug effects gene knockdown techniques

Bid and calpains cooperate to trigger oxaliplatin-induced apoptosis of cervical carcinoma HeLa cells. (2009)

Abstract The Bcl-2 homology 3-only protein Bid is an important mediator of death receptor-induced apoptosis. Recent reports and this study suggest that Bid may also mediate genotoxic drug-induced apoptosis of various human cancer cells. Here, we characterized the role of Bid and the mechanism of Bid activation during oxaliplatin-induced apoptosis of HeLa cervical cancer cells. Small hairpin RNA-mediated silencing of Bid protected HeLa cells against both death receptor- and oxaliplatin-induced apoptosis. Expression of a Bid mutant in which caspase-8 cleavage site was mutated (D59A) reactivated oxaliplatin-induced apoptosis in Bid-deficient cells but failed to reactivate death receptor-induced apoptosis, suggesting that caspase-8-mediated Bid cleavage did not contribute to oxaliplatin-induced apoptosis. Overexpression of bcl-2 or treatment with the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-dl-Asp-fluoromethylketone abolished caspase-2, -8, -9, and -3 activation as well as Bid cleavage in response to oxaliplatin, suggesting that Bid cleavage occurred downstream of mitochondrial permeabilization and was predominantly mediated by caspases. We also detected an early activation of calpains in response to oxaliplatin. Calpain inhibition reduced Bid cleavage, mitochondrial depolarization, and activation of caspase-9, -3, -2, and -8 in response to oxaliplatin. Further experiments, however, suggested that Bid cleavage by calpains was not a prerequisite for oxaliplatin-induced apoptosis: single-cell imaging experiments using a yellow fluorescent protein-Bid-cyan fluorescent protein probe demonstrated translocation of full-length Bid to mitochondria that was insensitive to calpain or caspase inhibition. Moreover, calpain inhibition showed a potent protective effect in Bid-silenced cells. In conclusion, our data suggest that calpains and Bid act in a cooperative, but mutually independent, manner to mediate oxaliplatin-induced apoptosis of HeLa cells.
Collections Ireland -> Royal College of Surgeons in Ireland -> PubMed

Full list of authors on original publication

Jochen H M Prehn, Donat Kögel, Markus Rehm, Caoimhín G Concannon, Frank E Murray, Mary D Cannon, Heiko Düssmann, Robert O'Byrne, Barbara Köhler, Sergio Anguissola

Experts in our system

1
Jochen H M Prehn
Royal College of Surgeons in Ireland
Total Publications: 206
 
2
Donat Kögel
Royal College of Surgeons in Ireland
Total Publications: 14
 
3
Markus Rehm
Royal College of Surgeons in Ireland
Total Publications: 55
 
4
Caoimhín G Concannon
Royal College of Surgeons in Ireland
Total Publications: 46
 
5
F E Murray
Royal College of Surgeons in Ireland
 
6
Mary Cannon
Royal College of Surgeons in Ireland
Total Publications: 90
 
7
Heiko Düssmann
Royal College of Surgeons in Ireland
Total Publications: 45