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Cordycepin enhances hyperthermia-induced apoptosis and cell cycle arrest by modulating the MAPK pathway in human lymphoma U937 cells

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Abstract

Background

Hyperthermia induces cancer cell death. However, the cytotoxic effect of hyperthermia is not sufficient. Cordycepin can also induce apoptosis in cancer cells and enhance the antitumoral activity of irradiation. To examine cordycepin-mediated enhancement of hyperthermia-induced apoptosis, this study investigated the combined effects and apoptotic mechanisms of hyperthermia and cordycepin on human leukemia U937 cells.

Methods

Cell viability and apoptosis were measured using MTT assays, Hoechst 33342 staining and Annexin V/PI double staining. The distribution of the cell cycle and sub-G1 phase, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were examined by flow cytometry. The expression of related proteins was analyzed by western blotting.

Results

Combined treatment with hyperthermia and cordycepin markedly augmented apoptosis by upregulating Bax and suppressing Bcl-2, Bid and activated caspase 3 and 8 expression, and apoptosis was decreased by Z-VAD-fmk (a pan caspase inhibitor). We also found that the MMP was significantly decreased and excessive ROS generation occurred. The combination treatment also induced arrest in the G2/M phase by downregulating cyclin dependent kinase 1 (CDK1) and cyclin B1 protein expression. Furthermore, it was observed that mitogen-activated protein kinase (MAPK) pathway including ERK, JNK and p38 signals was involved in the induction of apoptosis. The phosphorylated p38 and JNK were increased and ERK phosphorylation was decreased by the combined treatment. In addition, N-acetyl-l-cysteine (NAC) significantly protected the cells by restoring ROS levels and the activity of caspase-3, inactivating the MAPK pathway.

Conclusion

Cordycepin significantly enhanced hyperthermia-induced apoptosis and G2/M phase arrest in U937 cells. The combined treatment enhanced apoptosis through the MAPK pathway and mitochondrial dysfunction, and these effects could be rescued by NAC. We report for the first time that cordycepin can be used as a hyperthermia sensitizer to treat leukemia.

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Acknowledgements

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Funding

This study was funded by the Natural Science Foundation of Liaoning Province (Grant Number 2019-ZD-0564).

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Authors and Affiliations

  1. The School of Life Science and Biotechnology, Dalian University, Dalian, 116622, People’s Republic of China

    Liying Shi, He Cao, Siyu Fu, Zixian Jia, Xuan Lu & Dayong Yu

  2. Department of Environmental Health, University of Fukui School of Medical Science, University of Fukui, Eiheiji, 910-1193, Japan

    Zhengguo Cui

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  1. Liying Shi
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  2. He Cao
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  4. Zixian Jia
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  6. Zhengguo Cui
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  7. Dayong Yu
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Contributions

DY designed this study. LS, HC, SF, ZJ and XL performed the experiments. LS and HC wrote the manuscript and analyzed the results. ZC and DY planned and supervised the work. DY was responsible for funding acquisition. All authors read and approved the final manuscript.

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Correspondence to Zhengguo Cui or Dayong Yu.

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Shi, L., Cao, H., Fu, S. et al. Cordycepin enhances hyperthermia-induced apoptosis and cell cycle arrest by modulating the MAPK pathway in human lymphoma U937 cells. Mol Biol Rep 49, 8673–8683 (2022). https://doi.org/10.1007/s11033-022-07705-6

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