Advances in Clinical and Experimental Medicine
2018, vol. 27, nr 6, June, p. 833–840
Publication type: original article
Expression profiles of selected genes in tumors and matched surgical margins in oral cavity cancer: Do we have to pay attention to the molecular analysis of the surgical margins?
1 Department of Medical and Molecular Biology, School of Medicine with the Division of Dentistry in Zabrze, Medical University of Silesia in Katowice, Poland
2 Clinic of Oncological and Reconstructive Surgery, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice, Poland
3 Department of Statistics, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia in Katowice, Poland
Background. Head and neck squamous cell carcinomas (HNSCCs) are associated with an interplay between genetics and the environment; they account for 3% of all diagnosed malignant tumors in men and 2% of those in women.
Objectives. The aim of the study was to analyze the significance of TIMP3, SFRP1, SFRP2, CDH1, RASSF1, RORA, and DAPK1 gene expression in head and neck squamous cell carcinoma tumors, and in matching surgical margin samples. We also analyzed the association between clinical parameters and the expression of the selected genes.
Material and Methods. Following surgical resection, 56 primary HNSCC tumors and matching surgical margin samples were collected from patients at the Clinic of Oncological and Reconstructive Surgery of Maria Skłodowska-Curie Memorial Cancer Center and the Institute of Oncology in Gliwice, Poland. The gene expression levels were analyzed by quantitative reverse transcription (qRT)-PCR.
Results. SFRP1 gene expression was statistically significantly lower in the tumor samples than in the surgical margins (0.30 ±0.36 vs 0.62 ±0.36; p < 0.01). No correlation was found between gene expression and clinical parameters, except DAPK1, where low expression correlated with alcohol abuse (0.85 ±1.19 vs 1.97 ±3.22; p = 0.074). Moreover, patients with G3 grade tumors, i.e., poorly differentiated tumors, had significantly higher values of DAPK1 gene expression than the G1 (well-differentiated tumors) and G2 (moderately differentiated) groups.
Conclusion. There are many different reasons and concepts for altered gene expression in tumors and surgical margin tissue. Tumor heterogeneity and its microenvironment are undoubtedly linked to the biology of HNSCC. In order to understand specific tumor behavior and the microenvironment, further studies are needed. To find markers connected with cancer development and to provide insight into the earliest stages of cancer development, attention should also be focused on molecular analysis of the surgical margins.
gene expression, surgical margin, oral cavity, head and neck cancer
- Ignacio DN, Griffin JJ, Daniel MG, et al. An evaluation of treatment strategies for head and neck cancer in an African American population. West Indian Med J. 2013;62:504–509.
- Demokan S, Chuang A, Suoğlu Y, et al. Promoter methylation and loss of p16(INK4a) gene expression in head and neck cancer. Head Neck. 2012;34:1470–1475.
- Sang-Hyuk Lee SH, Lee NH, Jin SM, et al. Loss of heterozygosity of tumor suppressor genes (p16, Rb, E-cadherin, p53) in hypopharynx squamous cell carcinoma. Otolaryngol Head Neck Surg. 2011;145:64–70.
- Califano J, van der Riet P, Westra W, et al. Genetic progression model for head and neck cancer: Implications for field cancerization. Cancer Res. 1996;56:2488–2492.
- Slaughter DP, Southwick HW, Smejkal W. Field cancerization in oral stratified squamous epithelium: Clinical implications of multicentric origin. Cancer. 1953;6:963–968.
- Braakhuis BJ, Tabor MP, Kummer JA, Leemans CR, Brakenhoff RH. A genetic explanation of Slaughter’s concept of field cancerization: Evidence and clinical implications. Cancer Res. 2003;63:1727–1730.
- Angadi PV, Savitha JK, Rao SS, et al. Oral field cancerization: Current evidence and future perspectives. Oral Maxillofac Surg. 2012;16:171–180.
- Ushijima T. Epigenetic field for cancerization. J Biochem Mol Biol. 2007;40:142–150.
- Darnton SJ, Hardie LJ, Muc RS, et al. Tissue inhibitor of metalloproteinase-3 (TIMP-3) gene is methylated in the development of esophageal adenocarcinoma: Loss of expression correlates with poor prognosis. Int J Cancer. 2005;115:351–358.
- Frank B, Hoffmeister M, Klopp N, et al. Single nucleotide polymorphisms in Wnt signaling and cell death pathway genes and susceptibility to colorectal cancer. Carcinogenesis. 2010;8:1381–1386.
- Tremblay K, Daley D, Chamberland A, et al. Genetic variation in immune signaling genes differentially expressed in asthmatic lung tissues. J Allergy Clin Immunol. 2008;122:529–536.
- Kordi-Tamandani DM, Moazeni-Roodi AK, Rigi-Ladiz MA, et al. Promoter hypermethylation and expression profile of MGMT and CDH1 genes in oral cavity cancer. Arch Oral Biol. 2010;55:809–814.
- Oh HJ, Lee KK, Song SJ, et al. Role of the tumor suppressor RASSF1A in Mst1-mediated apoptosis. Cancer Res. 2006;66:2562–2569.
- Zhu Y, McAvoy S, Kuhn R, et al. RORA, a large common fragile site gene, is involved in cellular stress response. Oncogene. 2006;25:2901–2908.
- Lin Y, Hupp TR, Stevens C. Death-associated protein kinase (DAPK) and signal transduction: Additional roles beyond cell death. FEBS J. 2010;277:48–57.
- Brunner M, Ng BC, Veness MJ, et al. Comparison of the AJCC N staging system in mucosal and cutaneous squamous head and neck cancer. Laryngoscope. 2014;124:1598–1602.
- Rodrigues PC, Miguel MC, Bagordakis E, et al. Clinicopathological prognostic factors of oral tongue squamous cell carcinoma: A retrospective study of 202 cases. Int J Oral Maxillofac Surg. 2014;43:795–801.
- Marini A, Mirmohammadsadegh A, Nambiar S, et al. Epigenetic inactivation of tumor suppressor genes in serum of patients with cutaneous melanoma. J Invest Dermatol. 2006;126:422–431.
- Cao XL, Xu RJ, Zheng YY, et al. Expression of type IV collagen, metalloproteinase-2, metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in laryngeal squamous cell carcinomas. Asian Pac J Cancer Prev. 2011;12:3245–3249.
- McCormack A, Kaplan W, Gill AJ, et al. MGMT expression and pituitary tumors: Relationship to tumor biology. Pituitary. 2013;16:208–219.
- Zeng CH, Guo B, Chen J, et al. Expression profile of tumor suppressor gene RASSF1 in lacrimal gland carcinoma. Genet Mol Res. 2015;14:6993–6998.
- Belbin TJ, Singh B, Barber I, et al. Molecular classification of head and neck squamous cell carcinoma using cDNA microarrays. Cancer Res. 2002;62:1184–1190.
- El-Naggar AK, Kim HW, Clayman GL, et al. Differential expression profiling of head and neck squamous carcinoma: Significance in their phenotypic and biological classification. Oncogene. 2002;21:8206–8219.
- Maiti GP, Mondal P, Mukherjee N, et al. Overexpression of EGFR in head and neck squamous cell carcinoma is associated with inactivation of SH3GL2 and CDC25A genes. PLoS One. 2013;8:e63440.
- Schena M, Guarrera S, Buffoni L, et al. DNA repair gene expression level in peripheral blood and tumor tissue from non-small cell lung cancer and head and neck squamous cell cancer patients. DNA Repair (Amst). 2012;11:374–380.
- Stokes A, Joutsa J, Ala-Aho R, et al. Expression profiles and clinical correlations of degradome components in the tumor microenvironment of head and neck squamous cell carcinoma. Clin Cancer Res. 2010;16:2022–2035.
- Tsuchiya R, Yamamoto G, Nagoshi Y, et al. Expression of adenomatous polyposis coli (APC) in tumorigenesis of human oral squamous cell carcinoma. Oral Oncol. 2004;40:932–940.
- Dünne AA, Mandic R, Falkenberg S, et al. RT-PCR expression profiling of matrix metalloproteinases and their specific inhibitors in cell lines and fresh biopsies of squamous cell carcinomas of the head and neck. In Vivo. 2005;19:943–948.
- Sørensen BS, Toustrup K, Horsman MR, et al. Identifying pH independent hypoxia induced genes in human squamous cell carcinomas in vitro. Acta Oncol. 2010;49:895–905.
- Coskunpinar E, Oltulu YM, Orhan KS, et al. Identification of a differential expression signature associated with tumorigenesis and metastasis of laryngeal carcinoma. Gene. 2014;534:183–188.
- Fujii R, Imanishi Y, Shibata K, et al. Restoration of E-cadherin expression by selective Cox-2 inhibition and the clinical relevance of the epithelial-to-mesenchymal transition in head and neck squamous cell carcinoma. J Exp Clin Cancer Res. 2014;33:40.
- Ito S, Ohga T, Saeki H, et al. Promoter hypermethylation and quantitative expression analysis of CDKN2A (p14ARF and p16INK4a) gene in esophageal squamous cell carcinoma. Anticancer Res. 2007;27:3345–3353.
- Lo PH, Xie D, Chan KC, et al. Reduced expression of RASSF1A in esophageal and nasopharyngeal carcinomas significantly correlates with tumor stage. Cancer Lett. 2007;257:199–205.
- Pérez-Sayáns García M, Suárez-Peñaranda JM, Gayoso-Diz P, et al. Tissue inhibitor of metalloproteinases in oral squamous cell carcinomas – a therapeutic target? Cancer Lett. 2012;323:11–19.
- Burduk PK, Bodnar M, Sawicki P, et al. Expression of metalloproteinases 2 and 9 and tissue inhibitors 1 and 2 as predictors of lymph node metastases in oropharyngeal squamous cell carcinoma. Head Neck. 2015;37:418–422.
- Yu D, Zhou H, Xun Q, et al. microRNA-103 regulates the growth and invasion of endometrial cancer cells through the downregulation of tissue inhibitor of metalloproteinase 3. Oncol Lett. 2012;3:1221–1226.
- Rogler A, Kendziorra E, Giedl J, et al. Functional analyses and prognostic significance of SFRP1 expression in bladder cancer. J Cancer Res Clin Oncol. 2015;141:1779–1790.
- Sogabe Y, Suzuki H, Toyota M, et al. Epigenetic inactivation of SFRP genes in oral squamous cell carcinoma. Int J Oncol. 2008;32:1253–1261.
- Xiao C, Wang L, Zhu L, et al. Secreted frizzled related protein 2 is epigenetically silenced and functions as a tumor suppressor in oral squamous cell carcinoma. Mol Med Rep. 2014;10:2293–2298.
- Lee CH, Hung YJ, Lin CY, et al. Loss of SFRP1 expression is associated with aberrant beta-catenin distribution and tumor progression in mucoepidermoid carcinoma of salivary glands. Ann Surg Oncol. 2010;17:2237–2246.
- Hardisson D. Molecular pathogenesis of head and neck squamous cell carcinoma. Eur Arch Otorhinolaryngol. 2003;260:502–508.
- Fan CC, Wang TY, Cheng YA, et al. Expression of E-cadherin, Twist, and p53 and their prognostic value in patients with oral squamous cell carcinoma. J Cancer Res Clin Oncol. 2013;139:1735–1744.
- Burbee DG, Forgacs E, Zöchbauer-Müller S, et al. Epigenetic inactivation of RASSF1A in lung and breast cancers and malignant phenotype suppression. J Natl Cancer Inst. 2001;93:691–699.
- Righini CA, de Fraipont F, Timsit JF, et al. Tumor-specific methylation in saliva: A promising biomarker for early detection of head and neck cancer recurrence. Clin Cancer Res. 2007;13:1179–1185.
- Choudhury JH, Ghosh SK. Promoter hypermethylation profiling identifies subtypes of head and neck cancer with distinct viral, environmental, genetic and survival characteristics. PLoS One. 2015;10:e0129808.
- Ranhotra HS. The interplay between retinoic acid receptor-related orphan receptors and human diseases. J Recept Signal Transduct Res. 2012;32:181–189.
- Kottorou AE, Antonacopoulou AG, Dimitrakopoulos FI, et al. Altered expression of NFY-C and RORA in colorectal adenocarcinomas. Acta Histochem. 2012;114:553–561.
- Ye M, Li D, Zhou F, et al. Epigenetic regulation of death-associated protein kinase expression in primary gastric cancers from Chinese patients. Eur J Cancer Prev. 2012;21:241–246.
- Mariano FV, Rincon D, Gondak RO, et al. Carcinoma ex-pleomorphic adenoma of upper lip showing copy number loss of tumor suppressor genes. Oral Surg Oral Med Oral Pathol Oral Radiol. 2013;116:69–74.
- Tserga A, Michalopoulos NV, Levidou G, et al. Association of aberrant DNA methylation with clinicopathological features in breast cancer. Oncol Rep. 2012;27:1630–1638.
- Brait M, Loyo M, Rosenbaum E, et al. Correlation between BRAF mutation and promoter methylation of TIMP3, RARβ2 and RASSF1A in thyroid cancer. Epigenetics. 2012;7:710–719.
- Hashibe M, Boffetta P, Zaridze D, et al. Evidence for an important role of alcohol- and aldehyde-metabolizing genes in cancers of the upper aerodigestive tract. Cancer Epidemiol Biomarkers Prev. 2006;15:696–703.
- Erber R, Conradt C, Homann N, et al. TP53 DNA contact mutations are selectively associated with allelic loss and have a strong clinical impact in head and neck cancer. Oncogene. 1998;16:1671–1679.
- Hayashi M, Wu G, Roh JL, et al. Correlation of gene methylation in surgical margin imprints with locoregional recurrence in head and neck squamous cell carcinoma. Cancer. 2015;121:1957–1965.
- Wei DM, Liu DY, Lei DP, et al. Aberrant methylation and expression of DAPk1 in human hypopharyngeal squamous cell carcinoma. Acta Otolaryngol. 2015;135:70–78.
- Baylin SB, Ohm JE. Epigenetic gene silencing in cancer – a mechanism for early oncogenic pathway addiction? Nat Rev Cancer. 2006;6:107–116.
- Wong TS, Man MW, Lam AK, et al. The study of p16 and p15 gene methylation in head and neck squamous cell carcinoma and their quantitative evaluation in plasma by real-time PCR. Eur J Cancer. 2003;39:1881–1887.
- Leemans CR, Braakhuis BJ, Brakenhoff RH. The molecular biology of head and neck cancer. Nat Rev Cancer. 2011;11:9–22.
- Doorbar J. Molecular biology of human papillomavirus infection and cervical cancer. Clin Sci (Lond). 2006;110:525–541.
- Sritippho T, Pongsiriwet S, Lertprasertsuke N, et al. p16 – a possible surrogate marker for high-risk human papillomaviruses in oral cancer? Asian Pac J Cancer Prev. 2016;17:4049–4057.
- Garnaes E, Frederiksen K, Kiss K, et al. Double positivity for HPV DNA/p16 in tonsillar and base of tongue cancer improves prognostication: Insights from a large population-based study. Int J Cancer. 2016;139:2598–2605.
- Lim Y, Wan Y, Vagenas D, et al. Salivary DNA methylation panel to diagnose HPV-positive and HPV-negative head and neck cancers. BMC Cancer. 2016;16:749.
- Pavlik E, Nartova E, Astl J, et al. Detection of Helicobacter pylori and human papillomavirus in peroperative tissue biopsies collected from malignancies in oropharyngeal area. Am J Clin Exp Med. 2015;3:364–367.
- Braakhuis BJ, Tabor MP, Leemans CR, et al. Second primary tumors and field cancerization in oral and oropharyngeal cancer: Molecular techniques provide new insights and definitions. Head Neck. 2002;24:198–206.
- Perez-Ordoñez B, Beauchemin M, Jordan RC. Molecular biology of squamous cell carcinoma of the head and neck. J Clin Pathol. 2006;59:445–453.
- Tabor MP, Brakenhoff RH, van Houten VM, et al. Persistence of genetically altered fields in head and neck cancer patients: Biological and clinical implications. Clin Cancer Res. 2001;7:1523–1532.
- Braakhuis BJ, Tabor MP, Kummer JA, et al. A genetic explanation of Slaughter’s concept of field cancerization: Evidence and clinical implications. Cancer Res. 2003;63:1727–1730.
- Tabor MP, Brakenhoff RH, Ruijter-Schippers HJ, et al. Genetically altered fields as origin of locally recurrent head and neck cancer: A retrospective study. Clin Cancer Res. 2004;10:3607–3613.
- Szukała K, Brieger J, Bruch K, el al. Loss of heterozygosity on chromosome arm 13q in larynx cancer patients: Analysis of tumor, margin and clinically unchanged mucosa. Med Sci Monit. 2004;10:CR233–240.