Aberrant O-GlcNAc-modified proteins expressed in primary colorectal cancer

Phueaouan,Thanong; Chaiyawat,Parunya; Netsirisawan,Pukkavadee; Chokchaichamnankit,Daranee; Punyarit,Phaibul; Srisomsap,Chantragan; Svasti,Jisnuson; Champattanachai,Voraratt

doi:10.3892/or.2013.2794

Aberrant O-GlcNAc-modified proteins expressed in primary colorectal cancer

Authors:
- Thanong Phueaouan
- Parunya Chaiyawat
- Pukkavadee Netsirisawan
- Daranee Chokchaichamnankit
- Phaibul Punyarit
- Chantragan Srisomsap
- Jisnuson Svasti
- Voraratt Champattanachai
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Affiliations: Applied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, Thailand, Laboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailand, Department of Clinical Pathology, Army Institute of Pathology, Pramongkutklao Medical Center, Bangkok, Thailand
Published online on: October 11, 2013 https://doi.org/10.3892/or.2013.2794
Pages: 2929-2936

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Abstract

O-GlcNAcylation is a post-translational modification of serine and threonine residues which is dynamically regulated by 2 enzymes; O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) that catalyze the addition and removal of a single N-acetylglucosamine (GlcNAc) molecule, respectively. This modification is thought to be a nutrient sensor in highly proliferating cells via the hexosamine biosynthesis pathway, a minor branch of glycolysis. Although emerging evidence suggests that O-GlcNAc modification is associated with many types of cancer, identification of O-GlcNAc‑modified proteins and their role in cancer remain unexplored. In the present study, we demonstrated that O-GlcNAcylation is increased in primary colorectal cancer tissues, and that this augmentation is associated with an increased expression of OGT levels. Using 2-dimensional O-GlcNAc immunoblotting and LC-MS/MS analysis, 16 proteins were successfully identified and 8 proteins showed an increase in O-GlcNAcylation, including cytokeratin 18, heterogeneous nuclear ribonucleoproteins A2/B1 (hnRNP A2/B1), hnRNP H, annexin A2, annexin A7, laminin-binding protein, α-tubulin and protein DJ-1. Among these identified proteins, annexin A2 was further confirmed to show overexpression of O-GlcNAc in all cancer samples. The results, therefore, indicate that aberrant O-GlcNAcylation of proteins is associated with colorectal cancer and that identification of O-GlcNAc-modified proteins may provide novel biomarkers of cancer.

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1	Siegel R, Ward E, Brawley O and Jemal A: Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin. 61:212–236. 2011. View Article : Google Scholar : PubMed/NCBI
2	Attasara P: Hospital-Based Cancer Registry 2010. National Cancer Institute; Thailand, Bangkok: 2011
3	Siegel RL, Jemal A and Ward EM: Increase in incidence of colorectal cancer among young men and women in the United States. Cancer Epidemiol Biomarkers Prev. 18:1695–1698. 2009. View Article : Google Scholar : PubMed/NCBI
4	Gryfe R, Swallow C, Bapat B, Redston M, Gallinger S and Couture J: Molecular biology of colorectal cancer. Curr Probl Cancer. 21:233–300. 1997. View Article : Google Scholar
5	Gillies RJ, Robey I and Gatenby RA: Causes and consequences of increased glucose metabolism of cancers. J Nucl Med. 49(Suppl 2): S24–S42. 2008. View Article : Google Scholar : PubMed/NCBI
6	Chowdhury FU, Shah N, Scarsbrook AF and Bradley KM: (18F)FDG PET/CT imaging of colorectal cancer: a pictorial review. Postgrad Med J. 86:174–182. 2010. View Article : Google Scholar : PubMed/NCBI
7	Marshall S, Bacote V and Traxinger RR: Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system. Role of hexosamine biosynthesis in the induction of insulin resistance. J Biol Chem. 266:4706–4712. 1991.
8	Kreppel LK, Blomberg MA and Hart GW: Dynamic glycosylation of nuclear and cytosolic proteins. Cloning and characterization of a unique O-GlcNAc transferase with multiple tetratricopeptide repeats. J Biol Chem. 272:9308–9315. 1997. View Article : Google Scholar : PubMed/NCBI
9	Gao Y, Wells L, Comer FI, Parker GJ and Hart GW: Dynamic O-glycosylation of nuclear and cytosolic proteins: cloning and characterization of a neutral, cytosolic β-N-acetylglucosaminidase from human brain. J Biol Chem. 276:9838–9845. 2001.PubMed/NCBI
10	Hart GW, Slawson C, Ramirez-Correa G and Lagerlof O: Cross talk between O-GlcNAcylation and phosphorylation: roles in signaling, transcription, and chronic disease. Annu Rev Biochem. 80:825–858. 2011. View Article : Google Scholar : PubMed/NCBI
11	Mi W, Gu Y, Han C, et al: O-GlcNAcylation is a novel regulator of lung and colon cancer malignancy. Biochim Biophys Acta. 1812:514–519. 2011. View Article : Google Scholar : PubMed/NCBI
12	Gu Y, Mi W, Ge Y, et al: GlcNAcylation plays an essential role in breast cancer metastasis. Cancer Res. 70:6344–6351. 2010. View Article : Google Scholar : PubMed/NCBI
13	Champattanachai V, Netsirisawan P, Chaiyawat P, et al: Proteomic analysis and abrogated expression of O-GlcNAcylated proteins associated with primary breast cancer. Proteomics. 13:2088–2099. 2013.PubMed/NCBI
14	Pham VC, Henzel WJ and Lill JR: Rapid on-membrane proteolytic cleavage for Edman sequencing and mass spectrometric identification of proteins. Electrophoresis. 26:4243–4251. 2005. View Article : Google Scholar : PubMed/NCBI
15	Zhu Q, Zhou L, Yang Z, et al: O-GlcNAcylation plays a role in tumor recurrence of hepatocellular carcinoma following liver transplantation. Med Oncol. 29:985–993. 2012. View Article : Google Scholar : PubMed/NCBI
16	Lynch TP, Ferrer CM, Jackson SR, Shahriari KS, Vosseller K and Reginato MJ: Critical role of O-Linked β-N-acetylglucosamine transferase in prostate cancer invasion, angiogenesis, and metastasis. J Biol Chem. 287:11070–11081. 2012.
17	Yehezkel G, Cohen L, Kliger A, Manor E and Khalaila I: O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) in primary and metastatic colorectal cancer clones and effect of N-acetyl-β-D-glucosaminidase silencing on cell phenotype and transcriptome. J Biol Chem. 287:28755–28769. 2012. View Article : Google Scholar
18	Gerke V and Moss SE: Annexins: from structure to function. Physiol Rev. 82:331–371. 2002.PubMed/NCBI
19	Gerke V, Creutz CE and Moss SE: Annexins: linking Ca²⁺ signalling to membrane dynamics. Nat Rev Mol Cell Biol. 6:449–461. 2005. View Article : Google Scholar : PubMed/NCBI
20	Duncan R, Carpenter B, Main LC, Telfer C and Murray GI: Characterisation and protein expression profiling of annexins in colorectal cancer. Br J Cancer. 98:426–433. 2008. View Article : Google Scholar : PubMed/NCBI
21	Hsu PI, Huang MS, Chen HC, et al: The significance of ANXA7 expression and its correlation with poor cellular differentiation and enhanced metastatic potential of gastric cancer. J Surg Oncol. 97:609–614. 2008. View Article : Google Scholar : PubMed/NCBI
22	Ku NO and Omary MB: Identification and mutational analysis of the glycosylation sites of human keratin 18. J Biol Chem. 270:11820–11827. 1995. View Article : Google Scholar : PubMed/NCBI
23	Srikanth B, Vaidya MM and Kalraiya RD: O-GlcNAcylation determines the solubility, filament organization, and stability of keratins 8 and 18. J Biol Chem. 285:34062–34071. 2010. View Article : Google Scholar : PubMed/NCBI
24	Ku NO, Toivola DM, Strnad P and Omary MB: Cytoskeletal keratin glycosylation protects epithelial tissue from injury. Nat Cell Biol. 12:876–885. 2010. View Article : Google Scholar : PubMed/NCBI
25	Walgren JL, Vincent TS, Schey KL and Buse MG: High glucose and insulin promote O-GlcNAc modification of proteins, including α-tubulin. Am J Physiol Endocrinol Metab. 284:E424–E434. 2003. View Article : Google Scholar : PubMed/NCBI
26	Ji S, Kang JG, Park SY, Lee J, Oh YJ and Cho JW: O-GlcNAcylation of tubulin inhibits its polymerization. Amino Acids. 40:809–818. 2011. View Article : Google Scholar
27	Krecic AM and Swanson MS: hnRNP complexes: composition, structure, and function. Curr Opin Cell Biol. 11:363–371. 1999. View Article : Google Scholar : PubMed/NCBI
28	Martinez-Contreras R, Cloutier P, Shkreta L, Fisette JF, Revil T and Chabot B: hnRNP proteins and splicing control. Adv Exp Med Biol. 623:123–147. 2007. View Article : Google Scholar : PubMed/NCBI
29	Boukakis G, Patrinou-Georgoula M, Lekarakou M, Valavanis C and Guialis A: Deregulated expression of hnRNP A/B proteins in human non-small cell lung cancer: parallel assessment of protein and mRNA levels in paired tumour/non-tumour tissues. BMC Cancer. 10:4342010. View Article : Google Scholar : PubMed/NCBI
30	Fielding P, Turnbull L, Prime W, Walshaw M and Field JK: Heterogeneous nuclear ribonucleoprotein A2/B1 up-regulation in bronchial lavage specimens: a clinical marker of early lung cancer detection. Clin Cancer Res. 5:4048–4052. 1999.PubMed/NCBI
31	Garneau D, Revil T, Fisette JF and Chabot B: Heterogeneous nuclear ribonucleoprotein F/H proteins modulate the alternative splicing of the apoptotic mediator Bcl-x. J Biol Chem. 280:22641–22650. 2005. View Article : Google Scholar : PubMed/NCBI
32	Hope NR and Murray GI: The expression profile of RNA-binding proteins in primary and metastatic colorectal cancer: relationship of heterogeneous nuclear ribonucleoproteins with prognosis. Hum Pathol. 42:393–402. 2011. View Article : Google Scholar : PubMed/NCBI
33	Wang Z, Pandey A and Hart GW: Dynamic interplay between O-linked N-acetylglucosaminylation and glycogen synthase kinase-3-dependent phosphorylation. Mol Cell Proteomics. 6:1365–1379. 2007.
34	Gauczynski S, Peyrin JM, Haïk S, et al: The 37-kDa/67-kDa laminin receptor acts as the cell-surface receptor for the cellular prion protein. EMBO J. 20:5863–5875. 2001. View Article : Google Scholar : PubMed/NCBI
35	Wells L, Vosseller K, Cole RN, Cronshaw JM, Matunis MJ and Hart GW: Mapping sites of O-GlcNAc modification using affinity tags for serine and threonine post-translational modifications. Mol Cell Proteomics. 1:791–804. 2002.
36	Lefebvre T, Cieniewski C, Lemoine J, et al: Identification of N-acetyl-D-glucosamine-specific lectins from rat liver cytosolic and nuclear compartments as heat-shock proteins. Biochem J. 360:179–188. 2001.
37	Drummond FJ, Sowden J, Morrison K and Edwards YH: Colon carbonic anhydrase 1: transactivation of gene expression by the homeodomain protein Cdx2. FEBS Lett. 423:218–222. 1998. View Article : Google Scholar : PubMed/NCBI
38	Parkkila S, Rajaniemi H, Parkkila AK, et al: Carbonic anhydrase inhibitor suppresses invasion of renal cancer cells in vitro. Proc Natl Acad Sci USA. 97:2220–2224. 2000. View Article : Google Scholar : PubMed/NCBI
39	Zarghami N, Giai M, Yu H, et al: Creatine kinase BB isoenzyme levels in tumour cytosols and survival of breast cancer patients. Br J Cancer. 73:386–390. 1996. View Article : Google Scholar : PubMed/NCBI
40	Li T, Yang W, Li M, et al: Expression of selenium-binding protein 1 characterizes intestinal cell maturation and predicts survival for patients with colorectal cancer. Mol Nutr Food Res. 52:1289–1299. 2008. View Article : Google Scholar : PubMed/NCBI
41	Kim H, Kang HJ, You KT, et al: Suppression of human selenium-binding protein 1 is a late event in colorectal carcinogenesis and is associated with poor survival. Proteomics. 6:3466–3476. 2006. View Article : Google Scholar : PubMed/NCBI