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Deguelin, a selective silencer of the NPM1 mutant, potentiates apoptosis and induces differentiation in AML cells carrying the NPM1 mutation

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Abstract

Nucleophosmin (NPM1) is a multifunctional protein that functions as a molecular chaperone, shuttling between the nucleolus and the cytoplasm. In up to one third of patients with acute myeloid leukemia, mutation of NPM1 results in the aberrant cytoplasmic accumulation of mutant protein and is thought to be responsible for leukemogenesis. Deguelin, a rotenoid isolated from several plant species, has been shown to be a strong anti-tumor agent. Human leukemia cell lines were used for in vitro studies. Drug efficacy was evaluated by apoptosis and differentiation assays, and associated molecular events were assessed by Western blot. Gene silencing was performed using small interfering RNA (siRNA). Deguelin exhibited strong cytotoxic activity in the cell line of OCI-AML3 and selectively down-regulated the NPM1 mutant protein, which was accompanied by up-regulation of the activity of caspase-6 and caspase-8 in high concentrations. Deguelin induced differentiation of OCI-AML3 cells at a nontoxic concentration which was associated with a decrease in expression of activated caspase-8, p53, p21, and the 30-kD form of CCAAT/enhancer binding protein α (C/EBPα), whereas no effects were found in OCIM2 cells expressing NPM-wt. Moreover, treatment with siRNA in the NPM mutant cell line OCI-AML3 decreased expression of p53, p21, pro-caspase-8, and the 30-kD form of C/EBPα, and it inhibited proliferation and induced differentiation of the OCI-AML3 cells. In conclusion, deguelin is a potent in vitro inhibitor of the mutant form of NPM1, which provides the molecular basis for its anti-leukemia activities in NPM1 mutant acute myeloid leukemia cells.

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Acknowledgments

The authors would like to thank Prof. Brunangelo Falini, University of Perugia, for kindly providing the NPM1 mutation-specific antibody. The authors would also like to thank M.D. Minden (Ontario Cancer Institute, Toronto, ON, Canada) for providing the AML cell lines OCI-AML3 and OCIM2. This work was supported by the National Natural Science Foundation of China (no. 81070429 and no. 81372541).

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None declared

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Authors and Affiliations

  1. Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People’s Republic of China

    Sha Yi, Lu Wen, Jing He, Youping Wang, Fei Zhao, Jie Zhao, Zichu Zhao, Guohui Cui & Yan Chen

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  1. Sha Yi
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  2. Lu Wen
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  3. Jing He
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  8. Guohui Cui
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  9. Yan Chen
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Correspondence to Guohui Cui or Yan Chen.

Additional information

Sha Yi and Lu Wen contributed equally to this work.

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Fig. 1

Effect of deguelin on AML cells. (a) OCI-AML3 cells were treated with the indicated drugs for 48 h, and the protein levels of NPM-mut and NPM-wt were detected by western blot. γ-tubulin was used as a protein loading control. The histogram (b) shows the relative protein density of these proteins. (GIF 16 kb)

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Yi, S., Wen, L., He, J. et al. Deguelin, a selective silencer of the NPM1 mutant, potentiates apoptosis and induces differentiation in AML cells carrying the NPM1 mutation. Ann Hematol 94, 201–210 (2015). https://doi.org/10.1007/s00277-014-2206-x

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  • DOI: https://doi.org/10.1007/s00277-014-2206-x

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