title: Reactive oxygen species-regulated glycogen synthase kinase-3β activation contributes to all-trans retinoic acid-induced apoptosis in granulocyte-differentiated HL60 cells.
abstract: All-trans retionic acid (ATRA) treatment confers disease remission in acute promyelocytic leukemia (APL) patients by inducing granulocytic differentiation, which is followed by cell apoptosis. Although glycogen synthase kinase (GSK)-3β is known to be required for spontaneous cell death in neutrophils, the requirement of GSK-3β activation for the apoptotic effects remains unknown. This question is addressed in the present study using a model of ATRA-induced granulocytic differentiation and apoptosis in APL HL60 cells. ATRA at a therapeutic concentration (1 μM) induced granulocytic differentiation, followed by apoptosis. ATRA treatment caused decreased Mcl-1, caspase-3 activation, and PARP cleavage following the inactivation of phosphatidylinositol 3-kinase/AKT and the activation of GSK-3β. Pharmacologically and genetically inhibiting GSK-3β effectively retarded ATRA-induced Mcl-1 degradation and apoptosis. Additional differentiation inducers, phorbol 12-myristate 13-acetate and dimethyl sulfoxide, also triggered GSK-3β-dependent apoptosis. Mechanistically, ATRA caused the generation of reactive oxygen species (ROS) through increased expression of NADPH oxidase subunits (p47(phox) and p67(phox)) to facilitate ATRA-induced GSK-3β activation and cell apoptosis. This study indicates that ROS initiate GSK-3β-dependent apoptosis in granulocyte-differentiated cells after long-term ATRA treatment.
authors.affiliation: Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
authors.forename: Chi-Yun
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
authors.forename: Tsan-Tzu
authors.initials: TT
authors.lastname: Yang
authors.affiliation: Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
authors.forename: Chia-Ling
authors.initials: CL
authors.lastname: Chen
authors.affiliation: Institute of Internal Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
authors.forename: Wei-Chieh
authors.initials: WC
authors.lastname: Lin
authors.affiliation: Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan; Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan; Center of Infectious Disease and Signaling Research, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan. Electronic address: cflin@mail.ncku.edu.tw.
authors.forename: Chiou-Feng
authors.initials: CF
authors.lastname: Lin
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D006098:Granulocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018922:HL-60 Cells; D006801:Humans; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D017382:Reactive Oxygen Species; D014212:Tretinoin; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3
keywords: 6-bromoindirubin-3′-oxime (PubChem CID: 5287844); Acute promyelocytic leukemia; All-trans retionic acid; All-trans-retinoic acid (PubChem CID: 444795); Apoptosis; Diphenylene iodonium (PubChem CID: 3101); Glycogen synthase kinase-3β; LY294002 (PubChem CID: 3974); Lithium chloride (PubChem CID: 433294); NADPH oxidase; Phorbol 12-myristate 13-acetate (PubChem CID: 27924); Reactive oxygen species; SB415286 (PubChem CID: 4210951)