Decitabine, a DNA methyltransferase inhibitor, induces apoptosis in human leukemia cells through intracellular reactive oxygen species generation

Shin,Dong  Yeok; Park,You-Soo; Yang,Kwangmo; Kim,Gi-Young; Kim,Wun-Jae; Han,Min  Ho; Kang,Ho  Sung; Choi,Yung  Hyun

doi:10.3892/ijo.2012.1546

Decitabine, a DNA methyltransferase inhibitor, induces apoptosis in human leukemia cells through intracellular reactive oxygen species generation

Authors:
- Dong Yeok Shin
- You-Soo Park
- Kwangmo Yang
- Gi-Young Kim
- Wun-Jae Kim
- Min Ho Han
- Ho Sung Kang
- Yung Hyun Choi
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Affiliations: Department of Molecular Biology, Busan National University, Busan 609-735, Republic of Korea, Dongnam Institute of Radiological and Medicine Sciences, Busan 619-953, Republic of Korea, Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 690-756, Republic of Korea, Department of Urology, College of Medicine, Personalized Tumor Engineering Research Center, Chungbuk National University, Chungbuk 361-763, Republic of Korea, Department of Biomaterial Control (BK21 program), Graduate School, Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan 614-714, Republic of Korea, Department of Biochemistry, Dongeui University College of Oriental Medicine, Busan 614-052, Republic of Korea
Published online on: July 5, 2012 https://doi.org/10.3892/ijo.2012.1546
Pages: 910-918

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Abstract

The DNA methyltransferase inhibitor decitabine, 5-aza-2'-deoxycytidine, has been found to exert anti-metabolic and anticancer activities when tested against various cultured cancer cells. Furthermore, decitabine has been found to play critical roles in cell cycle arrest and apoptosis in various cancer cell lines; however, these roles are not well understood. In this study, we investigated decitabine for its potential anti-proliferative and apoptotic effects in human leukemia cell lines U937 and HL60. Our results indicated that treatment with decitabine resulted in significantly inhibited cell growth in a concentration- and time-dependent manner by the induction of apoptosis. Decitabine-induced apoptosis in U937 and HL60 cells was correlated with the downregulation of anti-apoptotic Bcl-2, XIAP, cIAP-1 and cIAP-2 protein levels, the cleavage of Bid proteins, the activation of caspases and the collapse of mitochondrial membrane potential (MMP). However, apoptosis induced by decitabine was attenuated by caspase inhibitors, indicating an important role for caspases in decitabine responses. The data further demonstrated that decitabine increased intracellular reactive oxygen species (ROS) generation. Moreover, N-acetyl-L-cysteine, a widely used ROS scavenger, effectively blocked the decitabine-induced apoptotic effects via inhibition of ROS production and MMP collapse. These observations clearly indicate that decitabine-induced ROS in human leukemia cells are key mediators of MMP collapse, which leads to apoptosis induction followed by caspase activation.

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