Gene expression profiling in bladder cancer identifies potential therapeutic targets

Hussain,Syed  A.; Palmer,Daniel  H.; Syn,Wing-Kin; Sacco,Joseph J.; Greensmith,Richard  M.D.; Elmetwali,Taha; Aachi,Vijay; Lloyd,Bryony  H.; Jithesh,Puthen  V.; Arrand,John; Barton,Darren; Ansari,Jawaher; Sibson,D.  Ross; James,Nicholas  D.

doi:10.3892/ijo.2017.3893

Gene expression profiling in bladder cancer identifies potential therapeutic targets

Authors:
- Syed A. Hussain
- Daniel H. Palmer
- Wing-Kin Syn
- Joseph J. Sacco
- Richard M.D. Greensmith
- Taha Elmetwali
- Vijay Aachi
- Bryony H. Lloyd
- Puthen V. Jithesh
- John Arrand
- Darren Barton
- Jawaher Ansari
- D. Ross Sibson
- Nicholas D. James
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Affiliations: Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool L69 3GA, UK, Regeneration and Repair Group, The Institute of Hepatology, Foundation of Liver Research, London SE5 9NT, UK, The Royal Liverpool and Broadgreen University Hospital Trust, Liverpool L7 8XP, UK, School of Cancer Sciences, University of Birmingham, Birmingham B15 2TT, UK, Beatson West Scotland Cancer Centre, Glasgow G12 0YN, UK
Published online on: March 2, 2017 https://doi.org/10.3892/ijo.2017.3893
Pages: 1147-1159
Copyright: © Hussain et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Despite advances in management, bladder cancer remains a major cause of cancer related complications. Characterisation of gene expression patterns in bladder cancer allows the identification of pathways involved in its pathogenesis, and may stimulate the development of novel therapies targeting these pathways. Between 2004 and 2005, cystoscopic bladder biopsies were obtained from 19 patients and 11 controls. These were subjected to whole transcript-based microarray analysis. Unsupervised hierarchical clustering was used to identify samples with similar expression profiles. Hypergeometric analysis was used to identify canonical pathways and curated networks having statistically significant enrichment of differentially expressed genes. Osteopontin (OPN) expression was validated by immunohistochemistry. Hierarchical clustering defined signatures, which differentiated between cancer and healthy tissue, muscle-invasive or non-muscle invasive cancer and healthy tissue, grade 1 and grade 3. Pathways associated with cell cycle and proliferation were markedly upregulated in muscle-invasive and grade 3 cancers. Genes associated with the classical complement pathway were downregulated in non-muscle invasive cancer. Osteopontin was markedly overexpressed in invasive cancer compared to healthy tissue. The present study contributes to a growing body of work on gene expression signatures in bladder cancer. The data support an important role for osteopontin in bladder cancer, and identify several pathways worthy of further investigation.

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1	IARC: Cancer incidence and mortality Worldwide. IARC CancerBase No. 10. Journal. 2015, http://globocan.iarc.fr.
2	Cancer Research UK: Bladder cancer statistics. 2015, http://www.cancerresearchuk.org/health-professional/cancer-statistics/statistics-by-cancer-type/bladder-cancer.
3	Kaufman DS, Shipley WU and Feldman AS: Bladder cancer. Lancet. 374:239–249. 2009. View Article : Google Scholar : PubMed/NCBI
4	Sylvester RJ, van der Meijden AP, Oosterlinck W, Witjes JA, Bouffioux C, Denis L, Newling DW and Kurth K: Predicting recurrence and progression in individual patients with stage Ta T1 bladder cancer using EORTC risk tables: a combined analysis of 2596 patients from seven EORTC trials. Eur Urol. 475–467. 2006.
5	Blaveri E, Simko JP, Korkola JE, Brewer JL, Baehner F, Mehta K, Devries S, Koppie T, Pejavar S, Carroll P, et al: Bladder cancer outcome and subtype classification by gene expression. Clin Cancer Res. 11:4044–4055. 2005. View Article : Google Scholar : PubMed/NCBI
6	Elsamman E, Fukumori T, Ewis AA, Ali N, Kajimoto K, Shinohara Y, Ishikawa M, Takahashi M, Nishitani MA, Baba Y, et al: Differences in gene expression between noninvasive and invasive transitional cell carcinoma of the human bladder using complementary deoxyribonucleic acid microarray: Preliminary results. Urol Oncol. 24:109–115. 2006. View Article : Google Scholar : PubMed/NCBI
7	Riester M, Taylor JM, Feifer A, Koppie T, Rosenberg JE, Downey RJ, Bochner BH and Michor F: Combination of a novel gene expression signature with a clinical nomogram improves the prediction of survival in high-risk bladder cancer. Clin Cancer Res. 18:1323–1333. 2012. View Article : Google Scholar : PubMed/NCBI
8	Sanchez-Carbayo M, Socci ND, Lozano J, Saint F and Cordon-Cardo C: Defining molecular profiles of poor outcome in patients with invasive bladder cancer using oligonucleotide microarrays. J Clin Oncol. 24:778–789. 2006. View Article : Google Scholar : PubMed/NCBI
9	Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U and Speed TP: Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 4:249–264. 2003. View Article : Google Scholar : PubMed/NCBI
10	Wu Z, Irizarry RA, Gentleman R, Martinez-Murillo F and Spencer F: A Model-based background adjustment for oligonucleotide expression arrays. J Am Stat Assoc. 99:909–917. 2004. View Article : Google Scholar
11	Bolstad BM, Collin F, Simpson KM, Irizarry RA and Speed TP: Experimental design and low-level analysis of microarray data. Int Rev Neurobiol. 60:25–58. 2004. View Article : Google Scholar : PubMed/NCBI
12	Pawitan Y, Michiels S, Koscielny S, Gusnanto A and Ploner A: False discovery rate, sensitivity and sample size for microarray studies. Bioinformatics. 21:3017–3024. 2005. View Article : Google Scholar : PubMed/NCBI
13	Huang W, Sherman BT and Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 4:44–57. 2009. View Article : Google Scholar
14	Edgar R, Domrachev M and Lash AE: Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res. 30:207–210. 2002. View Article : Google Scholar :
15	Dyrskjøt L, Kruhøffer M, Thykjaer T, Marcussen N, Jensen JL, Møller K and Ørntoft TF: Gene expression in the urinary bladder: A common carcinoma in situ gene expression signature exists disregarding histopathological classification. Cancer Res. 64:4040–4048. 2004. View Article : Google Scholar : PubMed/NCBI
16	Als AB, Dyrskjøt L, von der Maase H, Koed K, Mansilla F, Toldbod HE, Jensen JL, Ulhøi BP, Sengeløv L, Jensen KM, et al: Emmprin and survivin predict response and survival following cisplatin-containing chemotherapy in patients with advanced bladder cancer. Clin Cancer Res. 13:4407–4414. 2007. View Article : Google Scholar : PubMed/NCBI
17	Zhao WM and Fang G: Anillin is a substrate of anaphase-promoting complex/cyclosome (APC/C) that controls spatial contractility of myosin during late cytokinesis. J Biol Chem. 280:33516–33524. 2005. View Article : Google Scholar : PubMed/NCBI
18	Olakowski M, Tyszkiewicz T, Jarzab M, Krol R, Oczko-Wojciechowska M, Kowalska M, Kowal M, Gala GM, Kajor M, Lange D, et al: NBL1 and anillin (ANLN) genes over-expression in pancreatic carcinoma. Folia Histochem Cytobiol. 47:249–255. 2009. View Article : Google Scholar : PubMed/NCBI
19	Gruss OJ and Vernos I: The mechanism of spindle assembly: Functions of Ran and its target TPX2. J Cell Biol. 166:949–955. 2004. View Article : Google Scholar : PubMed/NCBI
20	Ronkainen H, Hirvikoski P, Kauppila S and Vaarala MH: Anillin expression is a marker of favourable prognosis in patients with renal cell carcinoma. Oncol Rep. 25:129–133. 2011.
21	Zaravinos A, Lambrou GI, Volanis D, Delakas D and Spandidos DA: Spotlight on differentially expressed genes in urinary bladder cancer. PLoS One. 6:e182552011. View Article : Google Scholar : PubMed/NCBI
22	Zhang Z, Zhang G and Kong C: High expression of polo-like kinase 1 is associated with the metastasis and recurrence in urothelial carcinoma of bladder. Urol Oncol. 31:1222–1230. 2013. View Article : Google Scholar
23	Stadler WM, Vaughn DJ, Sonpavde G, Vogelzang NJ, Tagawa ST, Petrylak DP, Rosen P, Lin CC, Mahoney J, Modi S, et al: An open-label, single-arm, phase 2 trial of the Polo-like kinase inhibitor volasertib (BI 6727) in patients with locally advanced or metastatic urothelial cancer. Cancer. 120:976–982. 2014. View Article : Google Scholar
24	Ahmed M, Behera R, Chakraborty G, Jain S, Kumar V, Sharma P, Bulbule A, Kale S, Kumar S, Mishra R, et al: Osteopontin: A potentially important therapeutic target in cancer. Expert. Opin Ther Targets. 15:1113–1126. 2011. View Article : Google Scholar
25	Xu ST, Guo C, Ding X, Fan WJ, Zhang FH, Xu WL and Ma YC: Role of osteopontin in the regulation of human bladder cancer proliferation and migration in T24 cells. Mol Med Rep. 11:3701–3707. 2015.PubMed/NCBI
26	Coppola D, Szabo M, Boulware D, Muraca P, Alsarraj M, Chambers AF and Yeatman TJ: Correlation of osteopontin protein expression and pathological stage across a wide variety of tumor histologies. Clin Cancer Res. 10:184–190. 2004. View Article : Google Scholar : PubMed/NCBI
27	Ke HL, Chang LL, Yang SF, Lin HH, Li CC, Wu DC and Wu WJ: Osteopontin overexpression predicts poor prognosis of upper urinary tract urothelial carcinoma. Urol Oncol. 29:703–709. 2011. View Article : Google Scholar
28	Zaravinos A, Volanis D, Lambrou GI, Delakas D and Spandidos DA: Role of the angiogenic components, VEGFA, FGF2, OPN and RHOC, in urothelial cell carcinoma of the urinary bladder. Oncol Rep. 28:1159–1166. 2012.PubMed/NCBI
29	Fang ZQ, Zang WD, Chen R, Ye BW, Wang XW, Yi SH, Chen W, He F and Ye G: Gene expression profile and enrichment pathways in different stages of bladder cancer. Genet Mol Res. 12:1479–1489. 2013. View Article : Google Scholar : PubMed/NCBI
30	Imano M, Satou T, Itoh T, Sakai K, Ishimaru E, Yasuda A, Peng YF, Shinkai M, Akai F, Yasuda T, et al: Immunohistochemical expression of osteopontin in gastric cancer. JJ Gastrointest Surg. 13:1577–1582. 2009. View Article : Google Scholar
31	Kita Y, Natsugoe S, Okumura H, Matsumoto M, Uchikado Y, Setoyama T, Owaki T, Ishigami S and Aikou T: Expression of osteopontin in oesophageal squamous cell carcinoma. Br J Cancer. 95:634–638. 2006. View Article : Google Scholar : PubMed/NCBI
32	Song JY, Lee JK, Lee NW, Yeom BW, Kim SH and Lee KW: Osteopontin expression correlates with invasiveness in cervical cancer. Aust N Z J Obstet Gynaecol. 49:434–438. 2009. View Article : Google Scholar : PubMed/NCBI
33	Forootan SS, Foster CS, Aachi VR, Adamson J, Smith PH, Lin K and Ke Y: Prognostic significance of osteopontin expression in human prostate cancer. Int J Cancer. 118:2255–2261. 2006. View Article : Google Scholar
34	Weber GF, Lett GS and Haubein NC: Osteopontin is a marker for cancer aggressiveness and patient survival. Br J Cancer. 103:861–869. 2010. View Article : Google Scholar : PubMed/NCBI
35	Collins AL, Rock J, Malhotra L, Frankel WL and Bloomston M: Osteopontin expression is associated with improved survival in patients with pancreatic adenocarcinoma. Ann Surg Oncol. 19:2673–2678. 2012. View Article : Google Scholar : PubMed/NCBI
36	Zhao L, Wang Y, Qu N, Huang C and Chen L: Significance of plasma osteopontin levels in patients with bladder urothelial carcinomas. Mol Diagn Ther. 16:311–316. 2012. View Article : Google Scholar : PubMed/NCBI
37	Naor D, Sionov RV and Ish-Shalom D: CD44: Structure, function, and association with the malignant process. Adv Cancer Res. 71:241–319. 1997. View Article : Google Scholar : PubMed/NCBI
38	Ang C, Chambers AF, Tuck AB, Winquist E and Izawa JI: Plasma osteopontin levels are predictive of disease stage in patients with transitional cell carcinoma of the bladder. BJU Int. 96:803–805. 2005. View Article : Google Scholar : PubMed/NCBI
39	Lim J and Thiery JP: Epithelial-mesenchymal transitions: Insights from development. Development. 139:3471–3486. 2012. View Article : Google Scholar : PubMed/NCBI
40	Baumgart E, Cohen MS, Silva Neto B, Jacobs MA, Wotkowicz C, Rieger-Christ KM, Biolo A, Zeheb R, Loda M, Libertino JA, et al: Identification and prognostic significance of an epithelialmesenchymal transition expression profile in human bladder tumors. Clin Cancer Res. 13:1685–1694. 2007. View Article : Google Scholar : PubMed/NCBI
41	Varela JC, Atkinson C, Woolson R, Keane TE and Tomlinson S: Upregulated expression of complement inhibitory proteins on bladder cancer cells and anti-MUC1 antibody immune selection. Int J Cancer. 123:1357–1363. 2008. View Article : Google Scholar : PubMed/NCBI
42	Babjuk M, Soukup V, Pesl M, Kostirova M, Drncova E, Smolova H, Szakacsova M, Getzenberg R, Pavlik I and Dvoracek J: Urinary cytology and quantitative BTA and UBC tests in surveillance of patients with pTapT1 bladder urothelial carcinoma. Urology. 71:718–722. 2008. View Article : Google Scholar : PubMed/NCBI
43	Cheng ZZ, Corey MJ, Pärepalo M, Majno S, Hellwage J, Zipfel PF, Kinders RJ, Raitanen M, Meri S and Jokiranta TS: Complement factor H as a marker for detection of bladder cancer. Clin Chem. 51:856–863. 2005. View Article : Google Scholar : PubMed/NCBI
44	Warzecha CC, Sato TK, Nabet B, Hogenesch JB and Carstens RP: ESRP1 and ESRP2 are epithelial cell-type-specific regulators of FGFR2 splicing. Mol Cell. 33:591–601. 2009. View Article : Google Scholar : PubMed/NCBI
45	Diez de Medina SG, Chopin D, El Marjou A, Delouvee A, LaRochelle WJ, Hoznek A, Abbou C, Aaronson SA, Thiery JP and Radvanyi F: Decreased expression of keratinocyte growth factor receptor in a subset of human transitional cell bladder carcinomas. Oncogene. 14:323–330. 1997. View Article : Google Scholar : PubMed/NCBI
46	Chaffer CL, Brennan JP, Slavin JL, Blick T, Thompson EW and Williams ED: Mesenchymal-to-epithelial transition facilitates bladder cancer metastasis: Role of fibroblast growth factor receptor-2. Cancer Res. 66:11271–11278. 2006. View Article : Google Scholar : PubMed/NCBI
47	Cancer Genome Atlas Research Network: Comprehensive molecular characterization of urothelial bladder carcinoma. Nature. 507:315–322. 2014. View Article : Google Scholar : PubMed/NCBI