Reactive oxygen species-regulated glycogen synthase kinase-3β activation contributes to all-trans retinoic acid-induced apoptosis in granulocyte-differentiated HL60 cells

Chi-Yun Wang; Tsan-Tzu Yang; Chia-Ling Chen; Wei-Chieh Lin; Chiou-Feng Lin

doi:10.1016/j.bcp.2013.12.021

Reactive oxygen species-regulated glycogen synthase kinase-3β activation contributes to all-trans retinoic acid-induced apoptosis in granulocyte-differentiated HL60 cells

Biochem Pharmacol. 2014 Mar 1;88(1):86-94. doi: 10.1016/j.bcp.2013.12.021. Epub 2014 Jan 6.

Authors

Chi-Yun Wang¹, Tsan-Tzu Yang¹, Chia-Ling Chen², Wei-Chieh Lin³, Chiou-Feng Lin⁴

Affiliations

¹ Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
² Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
³ Institute of Internal Medicine, College of Medicine, National Cheng Kung University, 1, University Road, Tainan 70101, Taiwan.
⁴ 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: [email protected].

PMID: 24406248
DOI: 10.1016/j.bcp.2013.12.021

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.

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).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apoptosis / drug effects*
Cell Differentiation / drug effects
Enzyme Activation / drug effects
Flow Cytometry
Glycogen Synthase Kinase 3 / metabolism*
Glycogen Synthase Kinase 3 beta
Granulocytes / drug effects
Granulocytes / metabolism
Granulocytes / pathology
HL-60 Cells
Humans
Reactive Oxygen Species / metabolism*
Tretinoin / pharmacology*

Substances

Reactive Oxygen Species
Tretinoin
GSK3B protein, human
Glycogen Synthase Kinase 3 beta
Glycogen Synthase Kinase 3