Calcium/calmodulin-dependent protein kinase inhibition potentiates thapsigargin-mediated cell death in SH-SY5Y human neuroblastoma cells.
We previously demonstrated a loss in Ca(2+)/Calmodulin-dependent protein kinase (CaM kinase) activity in SH-SY5Y undergoing thapsigargin-mediated apoptosis. To extend that finding we report that CaM kinase inhibition potentiates thapsigargin-mediated cell death. CaM kinase inhibitor KN93 on its own exhibits little toxicity up to 10 mM, as measured by release of lactate dehydrogenase (LDH) into the culture medium. In SH-SY5Y cells pretreated with KN93 and the non-selective protein kinase inhibitor k252a and then treated with 2 mM thapsigargin, loss of viability is significantly greater than in cells treated with thapsigargin alone. Pretreatment with the pan-caspase inhibitor Z-D-DCB prevented the thapsigargin-mediated increase in LDH release. Furthermore, thapsigargin-induced caspase-3-like activation, demonstrated by poly(ADP)ribose polymerase cleavage and pro-caspase-3 processing, was elevated in the presence of KN93.
McGinnis
K M
KM
Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Wang
K K
KK
Gnegy
M E
ME
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D001596:Benzylamines / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002227:Carbazoles / Q000494:pharmacology; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D026121:Indole Alkaloids; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000201:enzymology; D011480:Protease Inhibitors / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D019284:Thapsigargin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D000096926:Benzenesulfonamides
D001224:Aspartic Acid; D001596:Benzylamines; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002227:Carbazoles; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D011480:Protease Inhibitors; D013449:Sulfonamides; D019284:Thapsigargin; C093260:benzyloxycarbonyl-Asp-CH2OC(O)-2,6-dichlorobenzene; C072105:KN 93; D000096926:Benzenesulfonamides; C049985:staurosporine aglycone; C505407:CASP3 protein, human