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Deubiquitylatinase inhibitor b-AP15 induces c-Myc-Noxa-mediated apoptosis in esophageal squamous cell carcinoma

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Abstract

Esophageal squamous cell carcinoma (ESCC) is one of the most malignant tumors in east Asia. However, the molecular mechanism underlying its progression remains unclear. The ubiquitin–proteasome system (UPS) is a central mechanism for protein degradation and turnover. Accumulating evidence showed that more and more deubiquitinases could serve as attractive anti-cancer target. The expression of USP14 and UCH37 in esophagus squamous cell carcinoma tissues were examined by immunohistochemistry and western blot assays. Effect of b-AP15, a USP14 and UCH37 inhibitor, on ESCC cell growth was evaluated by cell viability assay. After cell lines being treated with b-AP15, cell cycle, apoptosis and the expression of related proteins were further explored to investigate the anti-ESCC mechanism of b-AP15. Results showed that deubiquitinating enzymes (DUBs) USP14 and UCH37 expressed at higher levels in ESCC tissues than in adjacent tissues. b-AP15 could inhibit cell proliferation and induce G2/M cell cycle arrest and apoptosis in ESCC cells. Mechanistically, b-AP15 treatment triggered Noxa-dependent apoptosis, which was regulated by c-Myc. Silencing Noxa and c-Myc could reduce b-AP15-induced apoptosis in ESCC cells. Our results revealed a novel mechanism of anti-tumor activity of b-AP15 in ESCC, and b-AP15 could be used as a potential therapeutic agent in ESCC.

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Data availability

Data sharing not applicable to this article as no datasets were generated or analyzed during the current study.

Abbreviations

ESCC:

Esophageal squamous cell cancer

UPS:

Ubiquitin–proteasome system

DUBs:

Deubiquitinating enzymes

USP14:

Ubiquitin-specific peptidase 14

UCH37:

Ubiquitin C-terminal hydrolase 37

IHC:

Immunohistochemistry

DAB:

3,3′-Diaminobenzidine tetrahydrochloride

ROS:

Reactive oxygen species

ER:

Endoplasmic reticulum

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Acknowledgements

And the authors would also like to thank those who contributed their tissue samples to support scientific research in medical fields.

Funding

This research was funded by the National Natural Science Foundation Grant of China (Grant Nos. 81001102, 81101894, 81672421, U1604189), Natural Science Foundation of Henan Province (Grant No. 162300410302), Outstanding Young Talent Research Fund of Zhengzhou University (Grant Nos. 51999223, 32210449) and Program for Science &Technology Innovation Talents in Universities of Henan Province (Grant No. 18HASTIT046).

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Author notes

  1. Beibei Sha and Xiaoyu Chen contributed equally to this work.

Authors and Affiliations

  1. College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China

    Beibei Sha, Xiaoyu Chen, Han Wu, Miaomiao Li, Longhao Wang, Xingge Liu, Ping Chen, Tao Hu & Pei Li

  2. Collaborative Innovation Center of Henan Province for Cancer Chemoprevention, Zhengzhou, 450001, China

    Beibei Sha, Xiaoyu Chen, Han Wu, Miaomiao Li, Longhao Wang, Xingge Liu, Ping Chen, Tao Hu & Pei Li

  3. Precision Medicine Center, Henan Institute of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, 450052, China

    Jianxiang Shi

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  1. Beibei Sha
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Contributions

Conceptualization, Tao Hu and Pei Li; Methodology, Beibei Sha, Xiaoyu Chen, Miaomiao Li and Longhao Wang; Validation, Beibei Sha, Xiaoyu Chen, Miaomiao Li, Longhao Wang and Han Wu; Results interpretation, Beibei Sha, Xiaoyu Chen, Miaomiao Li and Xingge Liu; Manuscript preparation: Ping Chen and Jianxiang Shi; Supervision, Tao Hu and Pei Li; Project administration, Tao Hu and Pei Li; Funding acquisition, Ping Chen, Tao Hu and Pei Li.

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Correspondence to Tao Hu or Pei Li.

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10495_2019_1561_MOESM1_ESM.pdf

Supplementary material 1 (PDF 1176 kb) Supplementary Figure. The effect of c-Myc on the accumulation of polyubiquitin. After transfected with the control siRNA, or c-Myc siRNA for 96 h, cell proteins were collected and knockdown efficiency and the effect of c-Myc on the accumulation of polyubiquitin were assessed by western blot analysis. GAPDH was used as loading control

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Sha, B., Chen, X., Wu, H. et al. Deubiquitylatinase inhibitor b-AP15 induces c-Myc-Noxa-mediated apoptosis in esophageal squamous cell carcinoma. Apoptosis 24, 826–836 (2019). https://doi.org/10.1007/s10495-019-01561-9

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