Dynamic changes in the gene expression profile during rat oral carcinogenesis induced by 4‑nitroquinoline 1‑oxide

Ge,Shuyun; Zhang,Ji; Du,Yanzhi; Hu,Bin; Zhou,Zengtong; Lou,Jianing

doi:10.3892/mmr.2016.4883

Dynamic changes in the gene expression profile during rat oral carcinogenesis induced by 4‑nitroquinoline 1‑oxide

Authors:
- Shuyun Ge
- Ji Zhang
- Yanzhi Du
- Bin Hu
- Zengtong Zhou
- Jianing Lou
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Affiliations: Department of Oral Mucosal Diseases, Shanghai Key Laboratory of Stomatology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China, State Key Laboratory of Medical Genomics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China, Department of Stomatology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, P.R. China
Published online on: February 8, 2016 https://doi.org/10.3892/mmr.2016.4883
Pages: 2561-2569
Copyright: © Ge et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The typical progression of oral cancer is from hyperplastic epithelial lesions through dysplasia to invasive carcinoma. It is important to investigate malignant oral cancer progression and development in order to determine useful approaches of prevention of dysplastic lesions. The present study aimed to gain insights into the underlying molecular mechanism of oral carcinogenesis by establishing a rat model of oral carcinogenesis using 4‑nitroquinoline 1‑oxide. Subsequently, transcription profile analysis using an integrating microarray was performed. The dynamic gene expression changes of the six stages of rat oral carcinogenesis (normal, mild epithelial dysplasia, moderate dysplasia, severe dysplasia, carcinoma in situ and oral squamous cell carcinomas) were analyzed using component plane presentations (CPP)‑self‑organizing map (SOM). Six genes were verified by quantitative polymerase chain reaction, immunohistochemistry and succinate dehydrogenase (SDH) activity assay kit. Numerous differentially expressed genes (DEGs) were identified during rat oral carcinogenesis. CPP‑SOM determined that these DEGs were primarily enriched during cell cycle, apoptosis, inflammatory response and tricarboxylic acid cycle, indicating the coordinated regulation of molecular networks. In addition, the expression of specific DEGs, such as janus kinase 3, cyclin‑dependent kinase A‑1, B‑cell chronic lymphocytic leukaemia/lymphoma 2‑like 2, nuclear factor‑κB, tumor necrosis factor receptor superfamily member 1A, cyclin D1 and SDH were identified to have high concordance with the results from microarray data. The current study demonstrated that oral carcinogenesis is a multi‑step and multi‑gene process, with a distinct pattern alteration along a continuum of malignant transformation. In addition, this comprehensive investigation provided a theoretical basis for the understanding of the molecular alterations associated with oral carcinogenesis.

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1	Ferlay J, Bray F, Parkin DM and Pisani P: Globocan 2000: Cancer Incidence and Mortality Worldwide. (IARC Cancer Bases No. 5). IARCPress; Lyon: 2001
2	Warnakulasuriya S: Global epidemiology of oral and oropha-ryngeal cancer. Oral Oncol. 45:309–316. 2009. View Article : Google Scholar
3	Kademani D, Bell RB, Schmidt BL, Blanchaert R, Fernandes R, Lambert P and Tucker WM; American Association of Oral and Maxillofacial Surgeons Task Force on Oral Cancer: A preliminary report from the American Association of Oral and Maxillofacial Surgeons Task Force on Oral Cancer. J Oral Maxillofac Surg. 66:2151–2157. 2008. View Article : Google Scholar : PubMed/NCBI
4	Ram H, Sarkar J, Kumar H, Konwar R, Bhatt ML and Mohammad S: Oral cancer: Risk factors and molecular pathogenesis. J Maxillofac Oral Surg. 10:132–137. 2011. View Article : Google Scholar :
5	Williams H: Molecular pathogenesis of oral squamous carcinoma. Mol Pathol. 53:165–172. 2000. View Article : Google Scholar : PubMed/NCBI
6	Nagpal JK and Das BR: Oral cancer: Reviewing the present understanding of its molecular mechanism and exploring the future directions for its effective management. Oral Oncol. 39:213–221. 2003. View Article : Google Scholar : PubMed/NCBI
7	Shaw R: The epigenetics of oral cancer. Int J Oral Maxillofac Surg. 35:101–108. 2006. View Article : Google Scholar
8	Scully C, Field JK and Tanzawa H: Genetic aberrations in oral or head and neck squamous cell carcinoma (SCCHN): 1. Carcinogen metabolism, DNA repair and cell cycle control. Oral Oncol. 36:256–263. 2000. View Article : Google Scholar : PubMed/NCBI
9	Todd R, Chou MY, Matossian K, Gallagher GT, Donoff RB and Wong DT: Cellular sources of transforming growth factor-alpha in human oral cancer. J Dent Res. 70:917–923. 1991. View Article : Google Scholar : PubMed/NCBI
10	Partridge M, Gullick WJ, Langdon JD and Sherriff M: Expression of epidermal growth factor receptor on oral squamous cell carcinoma. Br J Oral Maxillofac Surg. 26:381–389. 1988. View Article : Google Scholar : PubMed/NCBI
11	McDonald JS, Jones H, Pavelic ZP, Pavelic LJ, Stambrook PJ and Gluckman JL: Immunohistochemical detection of the H-ras, K-ras, and N-ras oncogenes in squamous cell carcinoma of the head and neck. J Oral Pathol Med. 23:342–346. 1994. View Article : Google Scholar : PubMed/NCBI
12	Williams HK, Sanders DS, Jankowski JA, Landini G and Brown AM: Expression of cadherins and catenins in oral epithelial dysplasia and squamous cell carcinoma. J Oral Pathol Med. 27:308–317. 1998. View Article : Google Scholar : PubMed/NCBI
13	Ravi D, Ramadas K, Mathew BS, Nalinakumari KR, Nair MK and Pillai MR: De novo programmed cell death in oral cancer. Histopathology. 34:241–249. 1999. View Article : Google Scholar : PubMed/NCBI
14	Duggan DJ, Bittner M, Chen Y, Meltzer P and Trent JM: Expression profiling using cDNA microarrays. Nat Genet. 21(Suppl): 10–14. 1999. View Article : Google Scholar : PubMed/NCBI
15	Kuo WP, Whipple ME, Sonis ST, Ohno-Machado L and Jenssen TK: Gene expression profiling by DNA microarrays and its application to dental research. Oral Oncol. 38:650–656. 2002. View Article : Google Scholar : PubMed/NCBI
16	Hwang D, Alevizos I, Schmitt WA, Misra J, Ohyama H, Todd R, Mahadevappa M, Warrington JA, Stephanopoulos G, Wong DT and Stephanopolus G: Genomic dissection for characterization of cancerous oral epithelium tissues using transcription profiling. Oral Oncol. 39:259–268. 2003. View Article : Google Scholar : PubMed/NCBI
17	Alevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M, Gallagher GT, Varvares M, Cohen D, Kim D, et al: Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis. Oncogene. 20:6196–6204. 2001. View Article : Google Scholar : PubMed/NCBI
18	Tang XH, Knudsen B, Bemis D, Tickoo S and Gudas LJ: Oral cavity and esophageal carcinogenesis modeled in carcinogen-treated mice. Clin Cancer Res. 10:301–313. 2004. View Article : Google Scholar : PubMed/NCBI
19	Kramer IR, Lucas RB, Pindborg JJ and Sobin LH: Definition of leukoplakia and related lesions: An aid to studies on oral precancer. Oral Surg Oral Med Oral Pathol. 46:518–539. 1978. View Article : Google Scholar : PubMed/NCBI
20	Horcajadas JA, Sharkey AM, Catalano RD, Sherwin JR, Domínguez F, Burgos LA, Castro A, Peraza MR, Pellicer A and Simón C: Effect of an intrauterine device on the gene expression profile of the endometrium. J Clin Endocrinol Metab. 91:3199–3207. 2006. View Article : Google Scholar : PubMed/NCBI
21	Xiao L, Wang K, Teng Y and Zhang J: Component plane presentation integrated self-organizing map for microarray data analysis. FEBS Lett. 538:117–124. 2003. View Article : Google Scholar : PubMed/NCBI
22	Dennis G Jr, Sherman BT, Hosack DA, Yang J, Gao W, Lane HC and Lempicki RA: DAVID: Database for annotation, visualization, and integrated discovery. Genome Biol. 4:32003. View Article : Google Scholar
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar
24	Evan GI and Vousden KH: Proliferation, cell cycle and apoptosis in cancer. Nature. 411:342–348. 2001. View Article : Google Scholar : PubMed/NCBI
25	Smith BD and Haffty BG: Molecular markers as prognostic factors for local recurrence and radioresistance in head and neck squamous cell carcinoma. Radiat Oncol Investig. 7:125–144. 1999. View Article : Google Scholar : PubMed/NCBI
26	Quelle DE, Ashmun RA, Shurtleff SA, Kato JY, Bar-Sagi D, Roussel MF and Sherr CJ: Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. Genes Dev. 7:1559–1571. 1993. View Article : Google Scholar : PubMed/NCBI
27	Miyamoto R, Uzawa N, Nagaoka S, Hirata Y and Amagasa T: Prognostic significance of cyclin D1 amplification and overexpression in oral squamous cell carcinomas. Oral Oncol. 39:610–618. 2003. View Article : Google Scholar : PubMed/NCBI
28	Robinson CM, Prime SS, Huntley S, Stone AM, Davies M, Eveson JW and Paterson IC: Overexpression of JunB in undifferentiated malignant rat oral keratinocytes enhances the malignant phenotype in vitro without altering cellular differentiation. Int J Cancer. 91:625–630. 2001. View Article : Google Scholar : PubMed/NCBI
29	Cheng YH, Li LA, Lin P, Cheng LC, Hung CH, Chang NW and Lin C: Baicalein induces G1 arrest in oral cancer cells by enhancing the degradation of cyclin D1 and activating AhR to decrease Rb phosphorylation. Toxicol Appl Pharmacol. 263:360–367. 2012. View Article : Google Scholar : PubMed/NCBI
30	Wilson GD, Saunders MI, Dische S, Richman PI, Daley FM and Bentzen SM: bcl-2 expression in head and neck cancer: An enigmatic prognostic marker. Int J Radial Oncol Biol Phys. 49:435–441. 2001. View Article : Google Scholar
31	May MJ and Ghosh S: Signal transduction through NF-kappaB. Immunol Today. 19:80–88. 1998. View Article : Google Scholar : PubMed/NCBI
32	Tracey L, Streck CJ, Du Z, Williams RF, Pfeffer LM, Nathwani AC and Davidoff AM: NF-kappaB activation mediates resistance to IFNbeta in MLL-rearranged acute lymphoblastic leukemia. Leukemia. 24:806–812. 2010. View Article : Google Scholar : PubMed/NCBI
33	Shostak K and Chariot A: NF-κB, stem cells and breast cancer: The links get stronger. Breast Cancer Res. 13:2142011. View Article : Google Scholar
34	Fan Y, Mao R and Yang J: NF-κB and STAT3 signaling pathways collaboratively link inflammation to cancer. Protein Cell. 4:176–185. 2013. View Article : Google Scholar : PubMed/NCBI
35	Thu YM, Su Y, Yang J, Splittgerber R, Na S, Boyd A, Mosse C, Simons C and Richmond A: NF-κB inducing kinase (NIK) modulates melanoma tumorigenesis by regulating expression of pro-survival factors through the β-catenin pathway. Oncogene. 31:2580–2592. 2012. View Article : Google Scholar :
36	Nakayama H, Ikebe T, Beppu M and Shirasuna K: High expression levels of nuclear factor kappa-B, IκB kinase α and Akt kinase in squamous cell carcinoma of the oral cavity. Cancer. 92:3037–3044. 2001. View Article : Google Scholar : PubMed/NCBI
37	Arnott CH, Scott KA, Moore RJ, Hewer A, Phillips DH, Parker P, Balkwill FR and Owens DM: Tumour necrosis factor-alpha mediates tumour promotion via a PKC alpha- and AP-1-dependent pathway. Oncogene. 21:4728–4738. 2002. View Article : Google Scholar : PubMed/NCBI
38	Rhodus NL, Ho V, Miller CS, Myers S and Ondrey F: NF-kappaB dependent cytokine levels in saliva of patients with oral preneoplastic lesions and oral squamous cell carcinoma. Cancer Detect Prev. 29:42–45. 2005. View Article : Google Scholar : PubMed/NCBI
39	Senthilnathan P, Padmavathi R, Magesh V and Sakthisekaran D: Modulation of TCA cycle enzymes and electron transport chain systems in experimental lung cancer. Life Sci. 78:1010–1014. 2006. View Article : Google Scholar
40	Selak MA, Armour SM, MacKenzie ED, Boulahbel H, Watson DG, Mansfield KD, Pan Y, Simon MC, Thompson CB and Gottlieb E: Succinate links TCA cycle dysfunction to oncogenesis by inhibiting HIF-alpha prolyl hydroxylase. Cancer Cell. 7:77–85. 2005. View Article : Google Scholar : PubMed/NCBI
41	Bayley JP, Devilee P and Taschner PE: The SDH mutation database: An online resource for succinate dehydrogenase sequence variants involved in pheochromocytoma, paraganglioma and mitochondrial complex II deficiency. BMC Med Genet. 6:392005. View Article : Google Scholar : PubMed/NCBI
42	Hao HX, Khalimonchuk O, Schraders M, Dephoure N, Bayley JP, Kunst H, Devilee P, Cremers CW, Schiffman JD, Bentz BG, et al: SDH5, a gene required for flavination of succinate dehydrogenase, is mutated in paraganglioma. Science. 325:1139–1142. 2009. View Article : Google Scholar : PubMed/NCBI
43	Ricketts C, Woodward ER, Killick P, Morris MR, Astuti D, Latif F and Maher ER: Germline SDHB mutations and familial renal cell carcinoma. J Natl Cancer Inst. 100:1260–1262. 2008. View Article : Google Scholar : PubMed/NCBI
44	Janeway KA, Kim SY, Lodish M, Nosé V, Rustin P, Gaal J, Dahia PL, Liegl B, Ball ER, Raygada M, et al NIH Pediatric and Wild-Type GIST Clinic: Defects in succinate dehydrogenase in gastrointestinal stromal tumors lacking KIT and PDGFRA mutations. Proc Natl Acad Sci USA. 108:314–318. 2011. View Article : Google Scholar :
45	Raimundo N, Baysal BE and Shadel GS: Revisiting the TCA cycle: Signaling to tumor formation. Trends Mol Med. 17:641–649. 2011. View Article : Google Scholar : PubMed/NCBI