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Autophagy blockade sensitizes human head and neck squamous cell carcinoma towards CYT997 through enhancing excessively high reactive oxygen species-induced apoptosis

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Abstract

The functional relationship between apoptosis and autophagy in anticancer drug treatment is extremely complex, and the molecular machinery is obscure. This study aims to investigate the efficacy of CYT997, a novel microtubule-disrupting agent, in head and neck squamous cell carcinomas (HNSCCs) and complete the autophagy-apoptosis puzzle involved in drug action. We report here that CYT997 exhibits anticancer activity by triggering oxidative stress-associated apoptosis in HNSCC cells. Interestingly, upregulation of autophagy by mTOR-dependent pathways appears to have a cytoprotective role in preventing apoptosis by inhibiting CYT997-induced excessively high levels of reactive oxygen species (ROS). Blockade of autophagy by ATG7 depletion or addition of autophagy inhibitor hydroxychloroquine (HCQ) sensitizes HNSCC cells to CYT997 as evidenced by enhanced ROS-associated apoptosis. Moreover, HCQ exhibits a good synergism with CYT997 on induction of apoptosis in HNSCC xenografts without cytotoxicity, suggesting combined treatment of CYT997 with autophagy inhibitors would increase the anticancer efficacy of CYT997. These findings unveil the importance of ROS in crosstalk between autophagy and apoptosis in CYT997 treatment, raising concerns that genetic or pharmacologic blockade of autophagy should be considered in the design of new therapeutics for HNSCC.

Key messages

• CYT997 exhibits anticancer activity by induction of ROS-associated apoptosis.

• mTOR-dependent cytoprotective autophagy prevents CYT997-induced apoptosis.

• Blockade of autophagy augments CYT997 efficacy by enhanced ROS-associated apoptosis.

• Combination of autophagy inhibitors with CYT997 is more effective against HNSCC.

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Abbreviations

3-MA:

3-methyladenine

CV:

cyclic voltammetry

DCFH-DA:

2′, 7′-dichlorodihydrofluorescein diacetate

HNSCC:

head and neck squamous cell carcinomas

HCQ:

hydroxychloroquine

IACUC:

Institutional Animal Care and Use Committee

IHC:

immunohistochemistry

JEOL:

JEM 1230 transmission electron microscope

NAC:

N-acetylcysteine

NaCAC:

sodium cacodylate

MVD:

microvessel density

O2 •− :

superoxide

p70S6K:

p70 ribosomal S6 kinase

ROS:

reactive oxygen species

SOD:

superoxide dismutase

TEM:

transmission electron microscopy

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Acknowledgements

This work was supported in part by Dental College of Georgia Special Funding Initiative and a grant from the Department of Defense (W81XWH-14-1-0412) (to Y.T.). We would like to thank the staff of the Electron Microscopy and Histology Core at Augusta University for assistance with EM data collection.

Funding

This work was supported in part by Dental College of Georgia Special Funding Initiative and a grant from the Department of Defense (W81XWH-14-1-0412) (to Y.T.).

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Authors and Affiliations

  1. School of Life Sciences, Chongqing University, Chongqing, China

    Lixia Gao

  2. Department of Oral Biology and Diagnostic Sciences, Augusta University, 1120 15th Street, Augusta, GA, 30912, USA

    Lixia Gao, Xiangdong Zhao, Liwei Lang, W. Andrew Yeudall & Yong Teng

  3. Department of Pediatrics, Emory Children’s Center, Emory University, Atlanta, GA, USA

    Chloe Shay

  4. The Robinson College of Business, Georgia State University, Atlanta, GA, USA

    Chloe Shay

  5. Georgia Cancer Center, Augusta University, Augusta, GA, USA

    W. Andrew Yeudall & Yong Teng

  6. Department of Biochemistry and Molecular Biology, Augusta University, Augusta, GA, USA

    Yong Teng

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  1. Lixia Gao
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Correspondence to Yong Teng.

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The study was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Augusta.

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The authors declare that they have no conflicts of interest.

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Gao, L., Zhao, X., Lang, L. et al. Autophagy blockade sensitizes human head and neck squamous cell carcinoma towards CYT997 through enhancing excessively high reactive oxygen species-induced apoptosis. J Mol Med 96, 929–938 (2018). https://doi.org/10.1007/s00109-018-1670-5

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  • DOI: https://doi.org/10.1007/s00109-018-1670-5

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