title: NADPH oxidase-derived reactive oxygen species are involved in the HL-60 cell monocytic differentiation induced by isoliquiritigenin.
abstract: The present study was undertaken to test the hypothesis that NADPH  oxidase-derived reactive oxygen species (ROS) are involved in isoliquiritigenin  (ISL)-induced monocytic differentiation in human acute promyelocytic leukemia HL-60 cells. Morphological changes, cell surface markers CD11b/CD14 and NBT-reducing ability were used to determine the differentiation of HL-60 cells, and 2,7-dichlorofluorescein  (DCFH-DA) was used to detect the level of intracellular ROS. ISL-induced  HL-60 cell differentiation was accompanied by an increase in the intracellular ROS  levels. l-Buthionine-(S,R)-sulfoximine (BSO), N-acetyl-l-cysteine (NAC), superoxide  dismutase (SOD) and 4-hydroxy-2,2,6,6-tetramethylpiperidinoxyl (Tempol) were used to interfere with ROS production. NADPH oxidase inhibitors, apocynin (APO) and diphenyleneiodonium (DPI) were used to study the role of NADPH oxidase in ISL-induced HL-60 cell differentiation. The ISL-induced HL-60 cell differentiation and intracellular ROS generation were enhanced by the oxidant BSO and inhibited by the antioxidants NAC, SOD, and tempol, and were also inhibited by the NADPH oxidase inhibitors APO and DPI. The protein and mRNA expression of the NADPH oxidase subunits gp91phox and p47phox were determined by Western blotting and RT-PCR, respectively. The levels of translation and transcription of the NADPH oxidase subunits gp91phox and p47phox increased markedly in a concentration-dependent manner. These findings suggest that NADPH oxidase plays a critical role in HL-60 cell differentiation induced by ISL and that NADPH oxidase-derived ROS is involved in the differentiation mechanism.
authors.affiliation: Key Laboratory of Xinjiang Endemic Phytomedicine Resources, Ministry of Education, School of Pharmacy, Shihezi University, Shihezi 832002, China. zqsyt@sohu.com
authors.forename: Hongmei
authors.initials: H
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Yao
authors.forename: Na
authors.initials: N
authors.lastname: Chen
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Chen
authors.forename: Hui
authors.initials: H
authors.lastname: Tian
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Wang
authors.forename: Qiusheng
authors.initials: Q
authors.lastname: Zheng
mesh_terms: D000975:Antioxidants / Q000494:pharmacology*; D018845:CD11 Antigens / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D053144:Cell Nucleus Shape / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D047188:Chalcones / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D018845:CD11 Antigens; D047188:Chalcones; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; C040920:isoliquiritigenin; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1