Effect of DJ-1 overexpression on the proliferation, apoptosis, invasion and migration of laryngeal squamous cell carcinoma SNU-46 cells through PI3K/AKT/mTOR

Wang,Bin; Qin,Hao; Wang,Yuejian; Chen,Weixiong; Luo,Jie; Zhu,Xiaolin; Wen,Weiping; Lei,Wenbin

doi:10.3892/or.2014.3286

Effect of DJ-1 overexpression on the proliferation, apoptosis, invasion and migration of laryngeal squamous cell carcinoma SNU-46 cells through PI3K/AKT/mTOR

Authors:
- Bin Wang
- Hao Qin
- Yuejian Wang
- Weixiong Chen
- Jie Luo
- Xiaolin Zhu
- Weiping Wen
- Wenbin Lei
View Affiliations

Affiliations: Otorhinolaryngology Hospital, Otorhinolaryngology Institute, The First Affiliated Hospital, Sun Yat-sen University, National Key Department of Otorhinolaryngology of People's Republic of China, Guangzhou, Guangdong 510080, P.R. China, Department of Otolaryngology, The First People's Hospital of Foshan, Foshan, Guangdong 528000, P.R. China
Published online on: June 24, 2014 https://doi.org/10.3892/or.2014.3286
Pages: 1108-1116

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to explore the effect of DJ-1-mediated PI3K/AKT/mTOR pathway on the proliferation, apoptosis, invasion, migration and other tumor biological characteristics of laryngeal squamous cell SNU-46, through stable transfection and overexpression of the DJ-1 gene. Retrovirus carrying DJ-1 gene was used to stabilize transfected human laryngeal squamous carcinoma SNU-46 cell line, and monoclonal cell line of stably overexpressed DJ-1 protein was screened out by G418. DJ-1 protein expression was determined by western blotting, and changes of p-AKT, p-mTOR and PTEN protein content were detected, followed by the detection of changes in proliferation, apoptosis, invasion, migration and other tumor biological characteristics of laryngeal squamous carcinoma cell line with stably transfected DJ-1 protein overexpression by flow cytometry, CCK-8 method and Transwell. We successfully constructed a laryngeal squamous carcinoma cell line of stably overexpressed DJ-1 protein and termed it SNU-46-DJ-1. After overexpression of DJ-1 protein, the levels of PTEN expression in laryngeal squamous cell SNU-46 decreased and p-AKT and p-mTOR protein expression levels increased. Compared to the untreated SNU-46 cells, the proliferation rate of SNU-46-DJ-1 cells increased (0.834±0.336 vs. 0.676±0.112; p<0.001); invasiveness was enhanced (165.7±13.6 vs. 100.0±17.4; p=0.001), the migration ability was enhanced (207.3±13.1 vs. 175.3±13.3; p=0.036), and the apoptosis rate decreased (3.533±5.167 vs. 16.397±5.447%; p=0.019). The overexpression of DJ-1 protein in laryngeal squamous carcinoma SNU-46 cells can accelerate proliferation rate, increase the invasion and migration capacity, and reduce apoptosis, by activating the PI3K/AKT/mTOR pathway.

View Figures

View References

1	Nagakubo D, Taira T, Kitaura H, et al: DJ-1, a novel oncogene which transforms mouse NIH3T3 cells in cooperation with ras. Biochem Biophys Res Commun. 231:509–513. 1997. View Article : Google Scholar : PubMed/NCBI
2	Annesi G, Savettieri G, Pugliese P, et al: DJ-1 mutations and parkinsonism-dementia-amyotrophic lateral sclerosis complex. Ann Neurol. 58:803–807. 2005. View Article : Google Scholar : PubMed/NCBI
3	Miller DW, Ahmad R, Hague S, et al: L166P mutant DJ-1, causative for recessive Parkinson’s disease, is degraded through the ubiquitin-proteasome system. J Biol Chem. 278:36588–36595. 2003.
4	Martinat C, Shendelman S, Jonason A, et al: Sensitivity to oxidative stress in DJ-1-deficient dopamine neurons: an ES-derived cell model of primary Parkinsonism. PLoS Biol. 2:e3272004. View Article : Google Scholar : PubMed/NCBI
5	Moore DJ, Zhang L, Troncoso J, et al: Association of DJ-1 and parkin mediated by pathogenic DJ-1 mutations and oxidative stress. Hum Mol Genet. 14:71–84. 2005. View Article : Google Scholar : PubMed/NCBI
6	Kim RH, Peters M, Jang Y, et al: DJ-1, a novel regulator of the tumor suppressor PTEN. Cancer Cell. 7:263–273. 2005. View Article : Google Scholar : PubMed/NCBI
7	MacKeigan JP, Clements CM, Lich JD, et al: Proteomic profiling drug-induced apoptosis in non-small cell lung carcinoma: identification of RS/DJ-1 and RhoGDIα. Cancer Res. 63:6928–6934. 2003.PubMed/NCBI
8	Hod Y: Differential control of apoptosis by DJ-1 in prostate benign and cancer cells. J Cell Biochem. 92:1221–1233. 2004. View Article : Google Scholar : PubMed/NCBI
9	Le Naour F, Misek DE, Krause MC, et al: Proteomics-based identification of RS/DJ-1 as a novel circulating tumor antigen in breast cancer. Clin Cancer Res. 7:3328–3335. 2001.PubMed/NCBI
10	Yoshino T, Shiina H, Urakami S, et al: Bcl-2 expression as a predictive marker of hormone-refractory prostate cancer treated with taxane-based chemotherapy. Clin Cancer Res. 12:6116–6124. 2006. View Article : Google Scholar : PubMed/NCBI
11	Shi W, Zhang X, Pintilie M, et al: Dysregulated PTEN-PKB and negative receptor status in human breast cancer. Int J Cancer. 104:195–203. 2003. View Article : Google Scholar : PubMed/NCBI
12	Davidson B, Hadar R, Schlossberg A, et al: Expression and clinical role of DJ-1, a negative regulator of PTEN, in ovarian carcinoma. Hum Pathol. 39:87–95. 2008. View Article : Google Scholar : PubMed/NCBI
13	Chu EA and Kim YJ: Laryngeal cancer: diagnosis and preoperative work-up. Otolaryngol Clin North Am. 41:673–695. 2008. View Article : Google Scholar : PubMed/NCBI
14	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar
15	Adelstein DJ, Li Y, Adams GL, et al: An intergroup phase III comparison of standard radiation therapy and two schedules of concurrent chemoradiotherapy in patients with unresectable squamous cell head and neck cancer. J Clin Oncol. 21:92–98. 2003. View Article : Google Scholar : PubMed/NCBI
16	Hoffman HT, Porter K, Karnell LH, et al: Laryngeal cancer in the United States: changes in demographics, patterns of care, and survival. Laryngoscope. 116:1–13. 2006. View Article : Google Scholar : PubMed/NCBI
17	Denis F, Garaud P, Bardet E, et al: Final results of the 94-01 French Head and Neck Oncology and Radiotherapy Group randomized trial comparing radiotherapy alone with concomitant radiochemotherapy in advanced-stage oropharynx carcinoma. J Clin Oncol. 22:69–76. 2004. View Article : Google Scholar
18	Forastiere AA, Maor M, Weber RS, et al: Long-term results of intergroup RTOG 91-11: a phase III trial to preserve the larynx-induction cisplatin/5-FU and radiation therapy versus concurrent cisplatin and radiation therapy versus radiation therapy. J Clin Oncol. 24(Suppl 18): 55172006.
19	Seiwert TY and Cohen EE: State-of-the-art management of locally advanced head and neck cancer. Br J Cancer. 92:1341–1348. 2005. View Article : Google Scholar : PubMed/NCBI
20	Brockstein B, Haraf DJ, Rademaker AW, et al: Patterns of failure, prognostic factors and survival in locoregionally advanced head and neck cancer treated with concomitant chemoradiotherapy: a 9-year, 337-patient, multi-institutional experience. Ann Oncol. 15:1179–1186. 2004.
21	Kutter J, Ozsahin M, Monnier P and Stupp R: Combined modality treatment with full-dose chemotherapy and concomitant boost radiotherapy for advanced head and neck carcinoma. Eur Arch Otorhinolaryngol. 262:1–7. 2005. View Article : Google Scholar : PubMed/NCBI
22	Kerr JF, Wyllie AH and Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 26:239–257. 1972. View Article : Google Scholar : PubMed/NCBI
23	Zhu XL, Wang ZF, Lei WB, et al: DJ-1: a novel independent prognostic marker for survival in glottic squamous cell carcinoma. Cancer Sci. 101:1320–1325. 2010. View Article : Google Scholar : PubMed/NCBI
24	Zhu XL, Wang ZF, Lei WB, et al: Tumorigenesis role and clinical significance of DJ-1, a negative regulator of PTEN, in supraglottic squamous cell carcinoma. J Exp Clin Cancer Res. 31:942012. View Article : Google Scholar : PubMed/NCBI
25	Sitaram RT, Cairney CJ, Grabowski P, et al: The PTEN regulator DJ-1 is associated with hTERT expression in clear cell renal cell carcinoma. Int J Cancer. 125:783–790. 2009. View Article : Google Scholar : PubMed/NCBI
26	Shendelman S, Jonason A, Martinat C, et al: DJ-1 is a redox-dependent molecular chaperone that inhibits α-synuclein aggregate formation. PLoS Biol. 2:e3622004.PubMed/NCBI
27	Vasseur S, Afzal S, Tardivel-Lacombe J, et al: DJ-1/PARK7 is an important mediator of hypoxia-induced cellular responses. Proc Natl Acad Sci USA. 106:1111–1116. 2009. View Article : Google Scholar : PubMed/NCBI
28	Lei W, Jia T, Su Z, et al: Combined effect of rapamycin and cisplatin on survival of Hep-2 cells in vitro. Oncol Res. 18:73–81. 2009. View Article : Google Scholar : PubMed/NCBI