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BIX-01294, a G9a inhibitor, suppresses cell proliferation by inhibiting autophagic flux in nasopharyngeal carcinoma cells

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Summary

G9a, a histone methyltransferase, has been found to be upregulated in a range of tumor tissues, and contributes to tumor growth and metastasis. However, the impact of G9a inhibition as a potential therapeutic target in nasopharyngeal carcinoma (NPC) is unclear. In the present study we aimed to investigate the anti-proliferative effect of G9a inhibition in the NPC cell lines CNE1 and CNE2, and to further elucidate the molecular mechanisms underlying these effects. The expression of G9a in NPC tumor tissues was significantly higher than that in normal nasopharyngeal tissues. The pharmacological inhibition of G9a by BIX-01294 (BIX) inhibited proliferation and induced caspase-independent apoptosis in NPC cells in vitro. Treatment with BIX induced autophagosome accumulation, which exacerbated the cytotoxic activity of BIX in NPC cells. Mechanistic studies have found that BIX impairs autophagosomes by initiating autophagy in a Beclin-1-independent way, and impairs autophagic degradation by inhibiting lysosomal cathepsin D activation, leading to lysosomal dysfunction. BIX was able to suppress tumor growth, possibly by inhibiting autophagic flux; it might therefore constitute a promising candidate for NPC therapy.

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Acknowledgements

The authors thank Editage (https://app.editage.cn/) for English language editing.

Funding

The Project was supported by grants from Chongqing Natural Science Foundation (Grant Serial Numbers: cstc2018jcyjAX0229) and Open Grants from the Key Laboratory of Electromagnetic Radiation Protection, Ministry of Education, China (No. 2017DCKF003).

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Authors and Affiliations

  1. Department of Otolaryngology Head and Neck Surgery, Xinqiao Hospital, Army Medical University (Third Military Medical University), Chongqing, China

    Qian Li, Liuqian Wang, Di Ji, Xiaomin Bao, Guojing Tan, Xiaojun Liang & Anchun Deng

  2. Department of Occupational Health, Army Medical University (Third Military Medical University), Chongqing, China

    Ping Deng, Huifeng Pi, Yonghui Lu, Chunhai Chen, Mindi He, Lei Zhang & Zhengping Yu

  3. Department of Environmental Medicine, Department of Emergency Medicine of First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Zhou Zhou

Authors
  1. Qian Li
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  2. Liuqian Wang
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  3. Di Ji
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  4. Xiaomin Bao
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  13. Zhou Zhou
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  15. Anchun Deng
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Contributions

Qian Li performed cell lines studies and wrote the manuscript. Liuqian Wang, Di Ji, Xiaomin Bao, Guojing Tan and Xiaojun Liang collected tissue samples and performed the bioinformatics analysis. Ping Deng, Huifeng Pi, Yonghui Lu, Chunhai Chen, Mindi He and Lei Zhang provided critical experimental technology and helped with data analysis. Zhou Zhou and Zhengping Yu were responsible for revision of the manuscript. Anchun Deng initiated the study, oversaw the progress of the project, and offered guidance.

Corresponding author

Correspondence to Anchun Deng.

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Conflict of interest

All authors declare that they have no conflict of interest. 

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee of Xinqiao Hospital, Army Medical University and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. 

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Informed consent was obtained from all individual participants included in the study.

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Supplementary Information

ESM 1

BIX-induced NPC cell viability inhibition does not occur by triggering apoptosis. (a) NPC cells were treated with different concentrations of BIX for 24 h. Apoptosis was then evaluated using flow cytometry, after double staining with annexin V and propidium iodide (PI). (b) The expression of cleaved caspase-3 and its substrate, cleaved PARP, was revealed using western blot analysis. A Jurkat cell total lysate was used as the positive control (PC) for the activation of caspase-3 and PARP. (c) NPC cells were treated with 10 μM BIX in the presence or absence of 20 μM Z-VAD for 24 h. Cell viability was determined using CCK-8 assay. Data are presented as mean ± SD. *P < 0.05, compared to the control. (PNG 4184 kb)

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Li, Q., Wang, L., Ji, D. et al. BIX-01294, a G9a inhibitor, suppresses cell proliferation by inhibiting autophagic flux in nasopharyngeal carcinoma cells. Invest New Drugs 39, 686–696 (2021). https://doi.org/10.1007/s10637-020-01053-7

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  • DOI: https://doi.org/10.1007/s10637-020-01053-7

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