title: Phenyl Saligenin Phosphate Induced Caspase-3 and c-Jun N-Terminal Kinase Activation in Cardiomyocyte-Like Cells.
abstract: At present, little is known about the effect(s) of organophosphorous compounds (OPs) on cardiomyocytes. In this study, we have investigated the effects of phenyl saligenin phosphate (PSP), two organophosphorothioate insecticides (diazinon and chlorpyrifos), and their acutely toxic metabolites (diazoxon and chlorpyrifos oxon) on mitotic and differentiated H9c2 cardiomyoblasts. OP-induced cytotoxicity was assessed by monitoring MTT reduction, LDH release, and caspase-3 activity. Cytotoxicity was not observed with diazinon, diazoxon, or chlorpyrifos oxon (48 h exposure; 200 μM). Chlorpyrifos-induced cytotoxicity was only evident at concentrations >100 μM. In marked contrast, PSP displayed pronounced cytotoxicity toward mitotic and differentiated H9c2 cells. PSP triggered the activation of JNK1/2 but not ERK1/2, p38 MAPK, or PKB, suggesting a role for this pro-apoptotic protein kinase in PSP-induced cell death. The JNK1/2 inhibitor SP 600125 attenuated PSP-induced caspase-3 and JNK1/2 activation, confirming the role of JNK1/2 in PSP-induced cytotoxicity. Fluorescently labeled PSP (dansylated PSP) was used to identify novel PSP binding proteins. Dansylated PSP displayed cytotoxicity toward differentiated H9c2 cells. 2D-gel electrophoresis profiles of cells treated with dansylated PSP (25 μM) were used to identify proteins fluorescently labeled with dansylated PSP. Proteomic analysis identified tropomyosin, heat shock protein β-1, and nucleolar protein 58 as novel protein targets for PSP. In summary, PSP triggers cytotoxicity in differentiated H9c2 cardiomyoblasts via JNK1/2-mediated activation of caspase-3. Further studies are required to investigate whether the identified novel protein targets of PSP play a role in the cytotoxicity of this OP, which is usually associated with the development of OP-induced delayed neuropathy.
authors.affiliation: School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: Shatha G
authors.initials: SG
authors.lastname: Felemban
authors.affiliation: School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: A Christopher
authors.initials: AC
authors.lastname: Garner
authors.affiliation: School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: Fathi A
authors.initials: FA
authors.lastname: Smida
authors.affiliation: John van Geest Cancer Research Centre, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: David J
authors.initials: DJ
authors.lastname: Boocock
authors.affiliation: School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Hargreaves
authors.affiliation: School of Science and Technology, Nottingham Trent University , Clifton Lane, Nottingham NG11 8NS, United Kingdom.
authors.forename: John M
authors.initials: JM
authors.lastname: Dickenson
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002800:Cholinesterase Inhibitors / Q000633:toxicity*; D058851:GPI-Linked Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D032386:Myoblasts, Cardiac / Q000187:drug effects* / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000633:toxicity*; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats
chemical_list: D000873:Anthracenes; D002800:Cholinesterase Inhibitors; D058851:GPI-Linked Proteins; D009943:Organophosphorus Compounds; D047428:Protein Kinase Inhibitors; C432165:pyrazolanthrone; C046357:phenylsaligenin cyclic phosphate; D048031:JNK Mitogen-Activated Protein Kinases; D000110:Acetylcholinesterase; C548246:Ache protein, rat; D053148:Caspase 3