Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

Zhe Liu; Douglas R. Hurst; Xing Qu; Li-Guang Lu; Chen-Zhou Wu; Yu-Yu Li; Yi Li

doi:10.1186/s40779-020-00275-3

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Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

RESEARCH | Updated：2023-02-28

- Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma
- Military Medical Research Vol. 8, Issue 2, Pages: 148-161(2021)
- Affiliations：
  
  1.State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  2.Department of Head and Neck Oncology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  3.Department of Pathology, University of Alabama at Birmingham, Birmingham, AL 35216, USA
  4.Department of Evidence Based Stomatology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
- Author bio：
  
  * liyi1012@163.com
- Funds：
- DOI：10.1186/s40779-020-00275-3
  CLC：
- Published：2021-06
- Accepted：
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Liu et al.: Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma. Mil Med Res, 2020, 7: 48.
DOI：

Liu et al.: Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma. Mil Med Res, 2020, 7: 48. DOI： 10.1186/s40779-020-00275-3.

摘要

Abstract

Background:

p53 and DIRAS3 are tumor suppressors that are frequently silenced in tumors. In this study

we sought to determine whether the concurrent re-expression of p53 and DIRAS3 could effectively induce head and neck squamous cell carcinoma (HNSCC) cell death.

Methods:

CAL-27 and SCC-25 cells were treated with Ad-DIRAS3 and rAd-p53 to induce re-expression of DIRAS3 and p53 respectively. The effects of DIRAS3 and p53 re-expression on the growth and apoptosis of HNSCC cells were examined by TUNEL assay

flow cytometric analysis and MTT. The effects of DIRAS3 and p53 re-expression on Akt phosphorylation

oncogene expression

and the interaction of 4E-BP1 with eIF4E were determined by real-time PCR

Western blotting and immunoprecipitation analysis. The ability of DIRAS3 and p53 re-expression to induce autophagy was evaluated by transmission electron microscopy

LC3 fluorescence microscopy and Western blotting. The effects of DIRAS3 and p53 re-expression on HNSCC growth were evaluated by using an orthotopic xenograft mouse model.

Results:

TUNEL assay and flow cytometric analysis showed that the concurrent re-expression of DIRAS3 and p53 significantly induced apoptosis (

0.001). MTT and flow cytometric analysis revealed that DIRAS3 and p53 reexpression significantly inhibited proliferation and induced cell cycle arrest (

0.001). Mechanistically

the concurrent re-expression of DIRAS3 and p53 down-regulated signal transducer and activation of transcription 3 (STAT3) and upregulated p21

WAF1/CIP1

and Bax (

0.001). DIRAS3 and p53 re-expression also inhibited Akt phosphorylation

increased the interaction of eIF4E with 4E-BP1

and reduced the expression of c-Myc

cyclin D1

vascular endothelial growth factor (VEGF)

fibroblast growth factor (FGF)

epidermal growth factor receptor (EGFR) and Bcl-2 (

0.001). Moreover

the concurrent re-expression of DIRAS3 and p53 increased the percentage of cells with GFP-LC3 puncta compared with that in cells treated with control adenovirus (50.00%±4.55%

. 4.67%±1.25%

0.001). LC3 fluorescence microscopy and Western blotting further showed that DIRAS3 and p53 re-expression significantly promoted autophagic activity but also inhibited autophagic flux

resulting in overall impaired autophagy. Finally

the concurrent re-expression of DIRAS3 and p53 significantly decreased the tumor volume compared with the control group in a HNSCC xenograft mouse model [(3.12±0.75) mm

. (189.02±17.54) mm

0.001].

Conclusions:

The concurrent re-expression of DIRAS3 and p53 is a more effective approach to HNSCC treatment than current treatment strategies.

关键词

Keywords

references

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