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NADPH Oxidase-Derived Reactive Oxygen Species Are Involved in the HL-60 Cell Monocytic Differentiation Induced by Isoliquiritigenin
Key Laboratory of Xinjiang Endemic Phytomedicine Resources, Ministry of Education, School of Pharmacy, Shihezi University, Shihezi 832002, China
Life Sciences School, Yantai University, Yantai 264005, China
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 24 September 2012; in revised form: 24 October 2012 / Accepted: 7 November 2012 / Published: 12 November 2012
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.
Keywords: HL-60 cell; isoliquiritigenin; differentiation; NADPH oxidase; reactive oxygen species
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MDPI and ACS Style
Chen, H.; Zhang, B.; Yao, Y.; Chen, N.; Chen, X.; Tian, H.; Wang, Z.; Zheng, Q. NADPH Oxidase-Derived Reactive Oxygen Species Are Involved in the HL-60 Cell Monocytic Differentiation Induced by Isoliquiritigenin. Molecules 2012, 17, 13424-13438.
Chen H, Zhang B, Yao Y, Chen N, Chen X, Tian H, Wang Z, Zheng Q. NADPH Oxidase-Derived Reactive Oxygen Species Are Involved in the HL-60 Cell Monocytic Differentiation Induced by Isoliquiritigenin. Molecules. 2012; 17(11):13424-13438.
Chen, Hongmei; Zhang, Bo; Yao, Ying; Chen, Na; Chen, Xiaoyu; Tian, Hui; Wang, Zhenhua; Zheng, Qiusheng. 2012. "NADPH Oxidase-Derived Reactive Oxygen Species Are Involved in the HL-60 Cell Monocytic Differentiation Induced by Isoliquiritigenin." Molecules 17, no. 11: 13424-13438.