The role of calpain in caspase activation during etoposide induced apoptosis in T cells.
T cells treated with the drug etoposide undergo apoptotic death characterized by early evidence of nuclear damage followed by loss of mitochondrial integrity and cell lysis. Calpains and caspases are cytoplasmic proteases and there is increasing evidence of cross-talk between these proteases in death pathways. In this study we have investigated the role of calpain, in etoposide-triggered apoptosis in the 2B4 murine T cell hybridoma. Cell permeable inhibitors of calpain, ALLnM, E64 and calpeptin that block Fas ligand-Fas-mediated death in T cells, blocked etoposide-induced nuclear damage, loss of mitochondrial integrity and cell lysis. A broad spectrum peptide inhibitor of caspases, ZVAD-fmk, partially blocked nuclear damage but poorly inhibited mitochondrial damage or cell lysis triggered by etoposide. Etoposide-induced expression of the cleaved, proteolytically active form of caspase 3, and DEVD-ase activity, detected prior to nuclear damage, were blocked in the presence of calpain inhibitors. Collectively, these data describe a role for calpain in regulating etoposide-induced apoptosis via a caspase-dependent pathway in T cells.
Varghese
J
J
National Centre for Biological Sciences, UAS-GKVK Campus, Bangalore, Karnataka, India.
Radhika
G
G
Sarin
A
A
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002154:Calpain / Q000037:antagonists & inhibitors / Q000502:physiology*; D020169:Caspases / Q000378:metabolism* / Q000502:physiology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D004789:Enzyme Activation; D005047:Etoposide / Q000494:pharmacology*; D053222:Fas Ligand Protein; D006825:Hybridomas; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D013601:T-Lymphocytes / Q000166:cytology* / Q000201:enzymology*
D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; C116501:acetyl-leucinal-leucinal-normethional; C071482:calpeptin; D005047:Etoposide; D002154:Calpain; D020169:Caspases; D007930:Leucine; C024974:E 64