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Isofraxidin, a potent reactive oxygen species (ROS) scavenger, protects human leukemia cells from radiation-induced apoptosis via ROS/mitochondria pathway in p53-independent manner

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Abstract

Ionizing radiation (IR) leads to oxidizing events such as excessive reactive oxygen species (ROS) in the exposed cells, resulting in further oxidative damage to lipids, proteins and DNA. To screen the potential radio-protective drug, the intracellular ROS was measured in irradiated U937 cells pretreated with 80 candidate traditional herbal medicine, respectively. Isofraxidin (IF) was one possible radio-protector in these 80 drugs. This study investigated the radio-protective role of IF, a Coumarin compound, in human leukemia cell lines, for the first time. Results indicate that IF protects against IR-induced apoptosis in U937 cells in the time- and concentration- dependent manner. IF decreases IR-induced intracellular ROS generation, especially hydroxyl radicals formation, inhibits IR-induced mitochondrial membrane potential loss and reduces IR-induced high intracellular Ca2+ levels regardless of ER stress. IF down-regulates the expression of caspase-3, phospho-JNK, phospho-p38 and activates Bax in mitochondria. IF inhibits cytochrome c release from mitochondria to cytosol. IF also moderates IR-induced Fas externalization and caspase-8 activation. IF also exhibits significant protection against IR-induced cell death in other leukemia cell lines such as Molt-4 cells and HL60 cells regardless of p53. Taken together, the data demonstrate that IF protects leukemia cells from radiation-induced apoptosis via ROS/mitochondria pathway in a p53-independent manner.

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Abbreviations

ROS:

Reactive oxygen species

IF:

Isofraxidin

IR:

Ionizing radiation

DMSO:

Dimethyl sulfoxide

MMP:

Mitochondrial membrane potential

TMRM:

Tetramethylrhodamine methyl ester

PI:

Propidium iodide

FITC:

Fluorescein isothiocyanate

HPF:

Hydroxyphenyl fluorescein

APF:

Aminophenyl fluorescein

HE:

Hydroethidine

DCFH:

Dichlorofluorescein diacetate

DAF-2 DA:

Diaminofluorescein-2 diacetate

pNA:

P-nitroanilide

IP3:

Inositol 1,4,5-trisphosphate

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Acknowledgments

This research was supported in part by a Grant-in-Aid for the Cooperative Research Project from Joint Usage/Research Center (Joint Usage/Research Center for Science-Based Natural Medicine) Institute of Natural Medicine. University of Toyama in 2013; in part by the International Exchange Fund of the Sugitani campus (Medical and Pharmacy) (A) in 2012, University of Toyama, Japan.

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Authors and Affiliations

  1. Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Sugitani 2630, Toyama, 930-0194, Japan

    Peng Li, Qing-Li Zhao, Paras Jawaid & Takashi Kondo

  2. Pathology Department, The 2nd Affiliated Hospital of Harbin Medical University, Nangang District, Harbin, 150081, China

    Li-Hua Wu & Yu-Fei Jiao

  3. Department of Gastrointestinal Pathophysiology, Institute of Natural Medicine, University of Toyama, Sugitani 2630, Toyama, 930-0194, Japan

    Makoto Kadowaki

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  1. Peng Li
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Correspondence to Qing-Li Zhao.

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Li, P., Zhao, QL., Wu, LH. et al. Isofraxidin, a potent reactive oxygen species (ROS) scavenger, protects human leukemia cells from radiation-induced apoptosis via ROS/mitochondria pathway in p53-independent manner. Apoptosis 19, 1043–1053 (2014). https://doi.org/10.1007/s10495-014-0984-1

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