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A novel synthetic novobiocin analog, FM-Nov17, induces DNA damage in CML cells through generation of reactive oxygen species

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Abstract

Objectives

To investigate the cytotoxicity of FM-Nov17 against chronic myeloid leukemia (CML) cells, we explored its underlying mechanisms mediating the induction of DNA damage and apoptotic cell death by reactive oxygen species (ROS).

Methods

MTT assays were used to measure the proliferation-inhibition ratio of K562 and K562/G01 cells. Flow cytometry (FCM) was used to test the level of extracellular ROS, DNA damage, cell cycle progression and apoptosis. Western blotting was used to verify the amount of protein.

Results

FM-Nov17 significantly inhibited the proliferation of K562 cells, with an IC50 of 58.28 ± 0.304 μM, and K562/G01 cells, with an IC50 of 62.36 ± 0.136 μM. FM-Nov17 significantly stimulated the generation of intracellular ROS, followed by the induction of DNA damage and the activation of the ATM–p53–r-H2AX pathway and checkpoint-related signals Chk1/Chk2, which led to increased numbers of cells in the S and G2/M phases of the cell cycle. Furthermore, FM-Nov17 induced apoptotic cell death by decreasing mitochondrial membrane potential and activating caspase-3 and PARP. The above effects were all prevented by the ROS scavenger N-acetylcysteine.

Conclusions

FM-Nov17-induces DNA damage and mitochondria-dependent cellular apoptosis in CML cells. The process is mediated by the generation of ROS.

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Authors and Affiliations

  1. Dept. of Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, PR China

    Nanwen Zhang, Lisen Huang, Jue Tian, Jianhua Xu & Lixian Wu

  2. Institute of Materia Medica, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, PR China

    Nanwen Zhang, Lisen Huang, Jue Tian, Jianhua Xu & Lixian Wu

  3. Fujian Key Laboratory of Natural Medicine Pharmacology, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, PR China

    Nanwen Zhang, Lisen Huang, Jue Tian, Jianhua Xu & Lixian Wu

  4. Department of Pharmacochemistry, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, PR China

    Fang Ke

  5. Department of Anatomy, School of Pharmacy, Fujian Medical University (FMU), Fuzhou, PR China

    Ming Zheng

  6. Union Hospital, FMU, Fujian Institute of Hematology, Fuzhou, PR China

    Xianling Chen

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  1. Nanwen Zhang
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  8. Lixian Wu
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Correspondence to Lixian Wu.

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Zhang, N., Huang, L., Tian, J. et al. A novel synthetic novobiocin analog, FM-Nov17, induces DNA damage in CML cells through generation of reactive oxygen species. Pharmacol. Rep 68, 423–428 (2016). https://doi.org/10.1016/j.pharep.2015.11.002

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  • DOI: https://doi.org/10.1016/j.pharep.2015.11.002

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