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N,N-Dimethyl phytosphingosine sensitizes HL-60/MX2, a multidrug-resistant variant of HL-60 cells, to doxorubicin-induced cytotoxicity through ROS-mediated release of cytochrome c and AIF

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Abstract

Doxorubicin (Dox) is widely used to treat a variety of tumors. However, resistance to this drug is common, making successful treatment more difficult. Previously, we introduced a novel phytosphingosine derivative, N,N-dimethyl phytosphingosine (DMPS), as a potent anticancer therapeutic agent in human leukemia cells. This study was performed to investigate whether DMPS can sensitize HL-60/MX2, a multidrug-resistant variant of HL-60, to Dox-induced apoptosis. Low concentrations of DMPS sensitized HL-60/MX2 cells to Dox-induced apoptosis. Combined Dox + DMPS treatment-induced apoptosis was accompanied by the activation of caspase-8 and caspase-3 as well as PARP cleavage. Cytochrome c and AIF release were also observed in Dox + DMPS-treated HL60/MX2 cells. Pretreatment with z-VAD-fmk markedly prevented caspase-3 activation and moderately suppressed apoptosis, suggesting that Dox + DMPS-induced apoptosis is somewhat (not completely) dependent on caspase. Cytochrome c and AIF release were not affected by pretreatment with z-VAD-fmk. The ROS scavenger NAC efficiently suppressed not only ROS generation, but also caspase-3-mediated PARP cleavage, apoptosis, and release of cytochrome c and AIF, indicating a role of ROS in combined Dox + DMPS treatment-induced apoptotic death signaling. Taken together, these observations suggest that DMPS may be used as a therapeutic agent for overcoming drug-resistance in cancer cells by enhancing drug-induced apoptosis.

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Abbreviations

AIF:

Apoptosis-inducing factor

DMPS:

N,N-Dimethyl phytosphingosine

Dox:

Doxorubicin

FACS:

Fluorescence-activated cell sorting

IAP:

Inhibitor of apoptosis proteins

MDR:

Multidrug resistance

MMP:

Mitochondrial membrane potential

NAC:

N-Acetylcysteine

pNA:

p-Nitroanilide

ROS:

Reactive oxygen species

S1P:

Sphingosine-1-phosphate

SK:

Sphingosine kinase

Tiron:

4,5-Dihydroxy-1,3-benzenedisulfonic acid

z-VAD-fmk:

N-Benzyloxycarbonyl-Val-Ala-Asp-fmk

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Acknowledgments

This work was supported by the National Nuclear R&D Program of the Ministry of Education, Science and Technology (MEST) of the Republic of Korea.

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Authors and Affiliations

  1. Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, 215-4 Gongneung-Dong, Nowon-Gu, Seoul, 139-706, Republic of Korea

    Byeong Mo Kim, Yun Jung Choi & Sung Hee Hong

  2. Department of Microbiology, Cornell University, Ithaca, NY, 14853-8101, USA

    Yong Heon Lee

  3. Cancer Research Institute and Department of Biomedical Science, College of Medicine, The Catholic University of Korea, Seoul, 137-701, Republic of Korea

    Young Ae Joe

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  1. Byeong Mo Kim
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  2. Yun Jung Choi
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  3. Yong Heon Lee
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  5. Sung Hee Hong
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Correspondence to Sung Hee Hong.

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Kim, B.M., Choi, Y.J., Lee, Y.H. et al. N,N-Dimethyl phytosphingosine sensitizes HL-60/MX2, a multidrug-resistant variant of HL-60 cells, to doxorubicin-induced cytotoxicity through ROS-mediated release of cytochrome c and AIF. Apoptosis 15, 982–993 (2010). https://doi.org/10.1007/s10495-010-0512-x

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