Calmodulin antagonists induce platelet apoptosis.
Calmodulin (CaM) antagonists induce apoptosis in various tumor models and inhibit tumor cell invasion and metastasis, thus some of which have been extensively used as anti-cancer agents. In platelets, CaM has been found to bind directly to the cytoplasmic domains of several platelet receptors. Incubation of platelets with CaM antagonists impairs the receptors-related platelet functions. However, it is still unknown whether CaM antagonists induce platelet apoptosis. Here we show that CaM antagonists N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W7), tamoxifen (TMX), and trifluoperazine (TFP) induce apoptotic events in human platelets, including depolarization of mitochondrial inner transmembrane potential, caspase-3 activation, and phosphatidylserine exposure. CaM antagonists did not incur platelet activation as detected by P-selectin surface expression and PAC-1 binding. However, ADP-, botrocetin-, and alpha-thrombin-induced platelet aggregation, platelet adhesion and spreading on von Willebrand factor surface were significantly reduced in platelets pre-treated with CaM antagonists. Furthermore, cytosolic Ca(2+) levels were obviously elevated by both W7 and TMX, and membrane-permeable Ca(2+) chelator BAPTA-AM significantly reduced apoptotic events in platelets induced by W7. Therefore, these findings indicate that CaM antagonists induce platelet apoptosis. The elevation of the cytosolic Ca(2+) levels may be involved in the regulation of CaM antagonists-induced platelet apoptosis.
Wang
Zhicheng
Z
School of Biological Science and Medical Engineering, Beihang University, Beijing, China.
Li
Suping
S
Shi
Quanwei
Q
Yan
Rong
R
Liu
Guanglei
G
Dai
Kesheng
K
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism*; D002147:Calmodulin / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000502:physiology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism*; D061945:Caspase Inhibitors; D004533:Egtazic Acid / Q000031:analogs & derivatives; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism / Q000494:pharmacology; D010718:Phosphatidylserines / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000378:metabolism / Q000494:pharmacology; D013629:Tamoxifen / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D014268:Trifluoperazine / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D014841:von Willebrand Factor / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology
D002147:Calmodulin; D002352:Carrier Proteins; D061945:Caspase Inhibitors; D019007:P-Selectin; D010718:Phosphatidylserines; D010975:Platelet Aggregation Inhibitors; D014841:von Willebrand Factor; D013629:Tamoxifen; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D014268:Trifluoperazine; D004533:Egtazic Acid; D013917:Thrombin; D053148:Caspase 3