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Arsenic Trioxide in APL

Involvement of CD95-independent caspase 8 activation in arsenic trioxide-induced apoptosis

Leukemia volume 14, pages 1743–1750 (2000)Cite this article

Abstract

Arsenic trioxide (As2O3)-treatment is effective in acute promyelocytic leukemia (APL) patients with t(15;17). Clinically achievable concentrations of As2O3 induce apoptosis in NB4, an APL cell line, in vitro. Here, to study the mechanism of As2O3-induced apoptosis, we established an As2O3-resistant subline, NB4/As. Growth of NB4/As was inhibited by 50% after 2 day-treatment (IC50) at 1.6 μM As2O3, whereas IC50 of NB4 was 0.3 μM. Degradation of PML-RARα and change of the PML-subcellular localization were similarly induced by As2O3 in NB4 and NB4/As, suggesting that their contribution to apoptosis is small. Treatment with 1 μM As2O3 induced the activation of caspase 3 as well as a loss of mitochondrial transmembrane potential (ΔΨm) in NB4 but not in NB4/As. Caspase 8 and Bid were also activated by As2O3 in NB4 but not in NB4/As. In NB4, an inhibitor of caspase 8 blocked not only the activation of caspase 3 but also the loss of ΔΨm. Neither cell line expressed CD95/Fas, and agonistic anti-Fas antibody (CH-11) failed to cause apoptosis. Neither antagonistic anti-CD95/Fas antibody nor anti-Fas ligand antibodies influenced the As2O3-induced apoptosis. NB4/As had a higher concentration of intracellular glutathione (GSH) than NB4 (96 vs 32 nmol/mg). Reduction of the GSH level by buthionine sulfoxide (BSO) completely restored the sensitivity to As2O3 in NB4/As. Furthermore, caspase activation and the loss of ΔΨm were recovered by combination treatment with BSO. These findings suggest that the As2O3 treatment activates caspase 8 in a CD95-independent but GSH concentration-dependent manner. In combination with BSO, As2O3 might be applied to therapy of leukemia/cancers which are insensitive to the clinically achievable concentrations of As2O3.

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Acknowledgements

We are grateful to Dr SJ Korsmeyer for providing anti-Bid antibody. This work was partly supported by a Grant-in-Aid from the Japanese Ministry of Education and Culture.

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  1. K Kitamura and Y Minami: KK and YM contributed equally to this work and should be regarded as cofirst authors

Authors and Affiliations

  1. Department of Infectious Diseases, Nagoya University School of Medicine, Nagoya, Japan

    K Kitamura, Y Minami, H Kiyoi & T Naoe

  2. First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

    Y Minami, K Yamamoto & H Saito

  3. Gifu International Institute of Biotechnology, Gifu, Japan

    Y Akao

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Kitamura, K., Minami, Y., Yamamoto, K. et al. Involvement of CD95-independent caspase 8 activation in arsenic trioxide-induced apoptosis. Leukemia 14, 1743–1750 (2000). https://doi.org/10.1038/sj.leu.2401900

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