title: Anandamide-induced Ca2+ elevation leading to p38 MAPK phosphorylation and subsequent cell death via apoptosis in human osteosarcoma cells.
abstract: The effect of anandamide on human osteoblasts is unclear. This study examined the effect of anandamide on viability, apoptosis, mitogen-activated protein kinases (MAPKs) and Ca2+ levels in MG63 osteosarcoma cells. Anandamide at 50-200 microM decreased cell viability via apoptosis as demonstrated by propidium iodide staining and activation of caspase-3. Immunoblotting suggested that anandamide induced expression of ERK, JNK and p38 MAPK. Anandamide-induced cell death and apoptosis were reversed by SB203580, but not by PD98059 and SP600125, suggesting that anandamide's action was via p38 MAPK, but not via ERK and JNK. Anandamide at 1-100 microM induced [Ca2+]i increases. Removal of extracellular Ca2+ decreased the anandamide response, indicating that anandamide induced Ca2+ influx and Ca2+ release. Chelation of intracellular Ca2+ with BAPTA reversed anandamide-induced cell death and p38 MAPK phosphorylation. Collectively, in MG63 cells, anandamide induced [Ca2+]i increases which evoked p38 MAPK phosphorylation. This p38 MAPK phosphorylation subsequently activated caspase-3 leading to apoptosis.
authors.affiliation: Department of Surgery, Kaohsiung Veterans General Hospital, Kaohsiung 813, Taiwan.
authors.forename: Shu-Shong
authors.initials: SS
authors.lastname: Hsu
authors.forename: Chun-Jen
authors.initials: CJ
authors.lastname: Huang
authors.forename: He-Hsiung
authors.initials: HH
authors.lastname: Cheng
authors.forename: Chiang-Ting
authors.initials: CT
authors.lastname: Chou
authors.forename: Hsiao-Ying
authors.initials: HY
authors.lastname: Lee
authors.forename: Jue-Long
authors.initials: JL
authors.lastname: Wang
authors.forename: I-Shu
authors.initials: IS
authors.lastname: Chen
authors.forename: Shiuh-Inn
authors.initials: SI
authors.lastname: Liu
authors.forename: Yih-Chau
authors.initials: YC
authors.lastname: Lu
authors.forename: Hong-Tai
authors.initials: HT
authors.lastname: Chang
authors.forename: Jong-Khing
authors.initials: JK
authors.lastname: Huang
authors.forename: Jin-Shyr
authors.initials: JS
authors.lastname: Chen
authors.forename: Chung-Ren
authors.initials: CR
authors.lastname: Jan
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001095:Arachidonic Acids / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism; D063385:Cannabinoid Receptor Modulators / Q000633:toxicity*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D063388:Endocannabinoids; D006801:Humans; D012516:Osteosarcoma; D010766:Phosphorylation; D053284:Polyunsaturated Alkamides / Q000633:toxicity*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D002614:Chelating Agents; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D048051:p38 Mitogen-Activated Protein Kinases; D053148:Caspase 3; D002118:Calcium; C078814:anandamide