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Lymphoma

2-Chloro-2′-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4

Leukemia volume 14, pages 299–306 (2000)Cite this article

Abstract

The mechanism of apoptosis induced by 2-chloro-2′-deoxyadenosine (2CdA) in human leukemia cell line MOLT-4 was investigated. 2CdA induced increases of 3′-OH ends of genomic DNA, ladder-like DNA fragmentation and phosphatidylserine translocation to the outer membrane, which are apoptotic characteristics. These apoptotic phenomena induced by 2CdA were inhibited by cycloheximide (CHX; a protein synthesis inhibitor), deoxycytidine (dC; a substrate of deoxycytidine kinase), acetyl Ile-Glu-Thr-Asp aldehyde (Ac-IETD-CHO; a caspase-8 inhibitor) and acetyl Asp-Glu-Val-Asp aldehyde (Ac-DEVD-CHO; a caspase-3 inhibitor). The protein synthesis-dependent expression of Fas and Fas ligand (Fas-L) was detected by treatment with 2CdA. The proteolytic processing of procaspases-8 and -3 to produce active fragments, caspases-8 (p18) and -3 (p17), respectively, was observed after treatment with 2CdA, and suppressed by cycloheximide. Increases in the activities of caspases-8 and -3 were observed after 2CdA treatment. Their activation was also dependent on protein synthesis. These results indicated that 2CdA-induced apoptosis was triggered by phosphorylation of 2CdA followed by the protein synthesis-dependent expression of Fas and Fas-L and activation of caspases-8 and -3.

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Acknowledgements

We thank Mr S Takagi for technical assistance on the bivariate flow cytometry for Annexin V/PI detection (Beckman Coulter K. K). This work was supported, in part, by Grants-in-Aid for Basic Scientific Research from the Ministry of Education, Science, Sports and Culture, Japan (No. 09660311 (OI), No. 09460133 (MK) and No. 08308032 (MK) and by Gakujutsu-Frontier Cooperative Research in Rakuno Gakuen University.

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  1. Laboratory of Radiation Biology, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan

    Y Nomura, O Inanami, K Takahashi & M Kuwabara

  2. Laboratory of Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan

    A Matsuda

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Nomura, Y., Inanami, O., Takahashi, K. et al. 2-Chloro-2′-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4. Leukemia 14, 299–306 (2000). https://doi.org/10.1038/sj.leu.2401649

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