Deubiquitinating enzyme USP20 is a positive regulator of Claspin and suppresses the malignant characteristics of gastric cancer cells

Wang,Chao; Yang,Chen; Ji,Jun; Jiang,Jinling; Shi,Min; Cai,Qu; Yu,Yingyan; Zhu,Zhenggang; Zhang,Jun

doi:10.3892/ijo.2017.3904

Deubiquitinating enzyme USP20 is a positive regulator of Claspin and suppresses the malignant characteristics of gastric cancer cells

Authors:
- Chao Wang
- Chen Yang
- Jun Ji
- Jinling Jiang
- Min Shi
- Qu Cai
- Yingyan Yu
- Zhenggang Zhu
- Jun Zhang
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Affiliations: Department of Oncology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, P.R. China, Department of Surgery, Shanghai Key Laboratory of Gastric Neoplasms, Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, P.R. China
Published online on: March 8, 2017 https://doi.org/10.3892/ijo.2017.3904
Pages: 1136-1146
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the clinical significance, the biological function and the mechanisms of USP20 in gastric cancer. The expression of USP20 in 89 pairs of primary gastric cancer and peritumoral gastric tissues specimens were measured by immunohistochemistry. The correlation of USP20 expression with the survival and the clinicopathological characteristics of patients were analyzed. Moreover, the underlying mechanisms of ectopic USP20 expression and its impact on GC cells were also investigated. We found that the expression of USP20 is relatively low in GC tissues and negatively correlated with tumor size, tumor invasion and TNM staging. High expression of USP20 in GC predicted longer survival. Experimentally, small interfering RNA-mediated knockdown of USP20 expression significantly promoted cell proliferation, accelerated G1-S phase transition and attenuated the autophagy activity. Overexpression of USP20 led to the inhibition of proliferation, G1-S cell cycle transition delay and autophagy activation. Mechanistically, we confirmed that silencing the expression of USP20 in GC cells could reduce Claspin protein levels without altering Claspin mRNA levels, which is involved in the antitumor activity of USP20. Furthermore, the expression level of Claspin was relatively higher in peritumoral tissue than that of GC tissues and higher expression of Claspin in GC was also correlated with good prognosis of patients. Given its pivotal role in gastric tumorigenesis and progression, USP20 functioned as the tumor suppressor in GC and possessed promising value to be a therapeutic target for GC.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Zeng H and Zhang S: The incidence and mortality of major cancers in China, 2012. Chin J Cancer. 35:732016. View Article : Google Scholar : PubMed/NCBI
3	Wang S and Yuan L: Predictive biomarkers for targeted and cytotoxic agents in gastric cancer for personalized medicine. Biosci Trends. 10:171–180. 2016. View Article : Google Scholar : PubMed/NCBI
4	McClurg UL and Robson CN: Deubiquitinating enzymes as oncotargets. Oncotarget. 6:9657–9668. 2015. View Article : Google Scholar : PubMed/NCBI
5	Park JJ, Yun JH and Baek KH: Polyclonal and monoclonal antibodies specific for ubiquitin-specific protease 20. Monoclon Antib Immunodiagn Immunother. 32:193–199. 2013.PubMed/NCBI
6	Nijman SM, Luna-Vargas MP, Velds A, Brummelkamp TR, Dirac AM, Sixma TK and Bernards R: A genomic and functional inventory of deubiquitinating enzymes. Cell. 123:773–786. 2005. View Article : Google Scholar : PubMed/NCBI
7	Fraile JM, Quesada V, Rodríguez D, Freije JM and Lopez-Otin C: Deubiquitinases in cancer: New functions and therapeutic options. Oncogene. 31:2373–2388. 2012. View Article : Google Scholar
8	Qu Q, Mao Y, Xiao G, Fei X, Wang J, Zhang Y, Liu J, Cheng G, Chen X, Wang J, et al: USP2 promotes cell migration and invasion in triple negative breast cancer cell lines. Tumour Biol. 36:5415–5423. 2015. View Article : Google Scholar : PubMed/NCBI
9	Cai JB, Shi GM, Dong ZR, Ke AW, Ma HH, Gao Q, Shen ZZ, Huang XY, Chen H, Yu DD, et al: Ubiquitin-specific protease 7 accelerates p14(ARF) degradation by deubiquitinating thyroid hormone receptor-interacting protein 12 and promotes hepatocellular carcinoma progression. Hepatology. 61:1603–1614. 2015. View Article : Google Scholar : PubMed/NCBI
10	Huang Z, Wen P, Kong R, Cheng H, Zhang B, Quan C, Bian Z, Chen M, Zhang Z, Chen X, Du X, et al: USP33 mediates Slit-Robo signaling in inhibiting colorectal cancer cell migration. Int J Cancer. 136:1792–1802. 2015. View Article : Google Scholar :
11	Kim SR, Kim JO, Lim KH, Yun JH, Han I and Baek KH: Regulation of pyruvate kinase isozyme M2 is mediated by the ubiquitin-specific protease 20. Int J Oncol. 46:2116–2124. 2015.PubMed/NCBI
12	Li Z, Wang D, Messing EM and Wu G: VHL protein-interacting deubiquitinating enzyme 2 deubiquitinates and stabilizes HIF-1alpha. EMBO Rep. 6:373–378. 2005. View Article : Google Scholar : PubMed/NCBI
13	Shanmugam I, Abbas M, Ayoub F, Mirabal S, Bsaili M, Caulder EK, Weinstock DM, Tomkinson AE, Hromas R and Shaheen M: Ubiquitin-specific peptidase 20 regulates Rad17 stability, checkpoint kinase 1 phosphorylation and DNA repair by homologous recombination. J Biol Chem. 289:22739–22748. 2014. View Article : Google Scholar : PubMed/NCBI
14	Yasunaga J, Lin FC, Lu X and Jeang KT: Ubiquitin-specific peptidase 20 targets TRAF6 and human T cell leukemia virus type 1 tax to negatively regulate NF-kappaB signaling. J Virol. 85:6212–6219. 2011. View Article : Google Scholar : PubMed/NCBI
15	Zhu M, Zhao H, Liao J and Xu X: HERC2/USP20 coordinates CHK1 activation by modulating CLASPIN stability. Nucleic Acids Res. 42:13074–13081. 2014. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Yuan J, Luo K, Deng M, Li Y, Yin P, Gao B, Fang Y, Wu P, Liu T and Lou Z: HERC2-USP20 axis regulates DNA damage checkpoint through Claspin. Nucleic Acids Res. 42:13110–13121. 2014. View Article : Google Scholar : PubMed/NCBI
18	Taillebourg E, Gregoire I, Viargues P, Jacomin AC, Thevenon D, Faure M and Fauvarque MO: The deubiquitinating enzyme USP36 controls selective autophagy activation by ubiquitinated proteins. Autophagy. 8:767–779. 2012. View Article : Google Scholar : PubMed/NCBI
19	Gossage L, Eisen T and Maher ER: VHL, the story of a tumour suppressor gene. Nat Rev Cancer. 15:55–64. 2015. View Article : Google Scholar
20	Kim RN, Kim A, Kim DW, Choi SH, Kim DS, Nam SH, Kang A, Kim MY, Park KH and Yoon BH: Analysis of indel variations in the human disease-associated genes CDKN2AIP, WDR66, USP20 and OR7C2 in a Korean population. J Genet. 91:e1–e11. 2012.PubMed/NCBI
21	Berthouze M, Venkataramanan V, Li Y and Shenoy SK: The deubiquitinases USP33 and USP20 coordinate beta2 adrenergic receptor recycling and resensitization. EMBO J. 28:1684–1696. 2009. View Article : Google Scholar : PubMed/NCBI
22	Bric A, Miething C, Bialucha CU, Scuoppo C, Zender L, Krasnitz A, Xuan Z, Zuber J, Wigler M, Hicks J, et al: Functional identification of tumor-suppressor genes through an in vivo RNA interference screen in a mouse lymphoma model. Cancer Cell. 16:324–335. 2009. View Article : Google Scholar : PubMed/NCBI
23	Li Z, Wang D, Na X, Schoen SR, Messing EM and Wu G: Identification of a deubiquitinating enzyme subfamily as substrates of the von Hippel-Lindau tumor suppressor. Biochem Biophys Res Commun. 294:700–709. 2002. View Article : Google Scholar : PubMed/NCBI
24	Yang CC, Suzuki M, Yamakawa S, Uno S, Ishii A, Yamazaki S, Fukatsu R, Fujisawa R, Sakimura K, Tsurimoto T, et al: Claspin recruits Cdc7 kinase for initiation of DNA replication in human cells. Nat Commun. 7:121352016. View Article : Google Scholar : PubMed/NCBI
25	Simicek M, Lievens S, Laga M, Guzenko D, Aushev VN, Kalev P, Baietti MF, Strelkov SV, Gevaert K, Tavernier J, et al: The deubiquitylase USP33 discriminates between RALB functions in autophagy and innate immune response. Nat Cell Biol. 15:1220–1230. 2013. View Article : Google Scholar : PubMed/NCBI
26	Lipinski MM, Hoffman G, Ng A, Zhou W, Py BF, Hsu E, Liu X, Eisenberg J, Liu J, Blenis J, et al: A genome-wide siRNA screen reveals multiple mTORCl independent signaling pathways regulating autophagy under normal nutritional conditions. Dev Cell. 18:1041–1052. 2010. View Article : Google Scholar : PubMed/NCBI
27	Kommaddi RP, Jean-Charles PY and Shenoy SK: Phosphorylation of the deubiquitinase USP20 by protein kinase A regulates post-endocytic trafficking of β2 adrenergic receptors to autophagosomes during physiological stress. J Biol Chem. 290:8888–8903. 2015. View Article : Google Scholar : PubMed/NCBI