Skip to main content
Top
Published in: Annals of Surgical Oncology 8/2013

01-08-2013 | Translational Research and Biomarkers

Molecular Staging of Surgical Margins in Oral Squamous Cell Carcinoma Using Promoter Methylation of p16INK4A, Cytoglobin, E-cadherin, and TMEFF2

Authors: Richard J. Shaw, MD, Andrew J. Hobkirk, MBChB, BDS, George Nikolaidis, PhD, Julia A. Woolgar, PhD, Asterios Triantafyllou, PhD, James S. Brown, MD, Triantafillos Liloglou, PhD, Janet M. Risk, PhD

Published in: Annals of Surgical Oncology | Issue 8/2013

Login to get access

Abstract

Background

Local recurrence in oral squamous cell carcinoma (OSCC) despite clear surgical margins may indicate the presence of residual, sub-microscopic disease. Molecular assessment of surgical margins may provide a greater prognostic sensitivity compared to histopathology. We aimed to determine whether promoter methylation in deep and mucosal resection margins can predict recurrence in OSCC.

Methods

Forty-eight consecutive OSCC cases were recruited and a 5 mm3 tumor sample plus 5 deep and 5 mucosal margin samples were snap frozen. Clinical, pathological, adjuvant therapy, and outcome data were recorded. Tumors were informative if >5 % promoter methylation was found for ≥1 of 4 genes using qMSP. Margins were declared molecularly positive if >1 % promoter methylation was found in any margin.

Results

Thirty (63 %) of 48 cases were methylation informative. Mucosal margin samples were largely positive for methylation (26 of 30, 87 %), indicating the presence of field cancerization. Methylation at ≥1 gene promoters in ≥1 deep margin correlated with the presence of close/involved mucosal margins (P = 0.027) and increased pT status (P = 0.027) but not the status of deep margins, recurrence, or survival.

Conclusions

The current gene panel did not add prognostic information to histopathological reporting of resection margins. Future efforts should concentrate on improving gene selection, informativity, and assay performance in the patient group with intermediate indications for adjuvant therapy.
Appendix
Available only for authorised users
Literature
2.
go back to reference Weijers M, Snow GB, Bezemer DP, van dr Wal JE, van der Waal I. The status of the deep surgical margins in tongue and floor of mouth squamous cell carcinoma and risk of local recurrence; an analysis of 68 patients. Int J Oral Maxillofac Surg. 2004;33:146.9. Weijers M, Snow GB, Bezemer DP, van dr Wal JE, van der Waal I. The status of the deep surgical margins in tongue and floor of mouth squamous cell carcinoma and risk of local recurrence; an analysis of 68 patients. Int J Oral Maxillofac Surg. 2004;33:146.9.
3.
go back to reference Braakhuis BJ, Bloemena E, Leemans CR, Brakenhoff RH. Molecular analysis of surgical margins in head and neck cancer: more than a marginal issue. Oral Oncol. 2010;46:485.91. Braakhuis BJ, Bloemena E, Leemans CR, Brakenhoff RH. Molecular analysis of surgical margins in head and neck cancer: more than a marginal issue. Oral Oncol. 2010;46:485.91.
4.
go back to reference Sutton DN, Brown JS, Rogers SN, Vaughan ED, Woolgar JA. The prognostic implications of the surgical margin in oral squamous cell carcinoma. Int J Oral Maxillofac Surg. 2003;32:30.4. Sutton DN, Brown JS, Rogers SN, Vaughan ED, Woolgar JA. The prognostic implications of the surgical margin in oral squamous cell carcinoma. Int J Oral Maxillofac Surg. 2003;32:30.4.
5.
go back to reference Rogers SN, Brown JS, Woolgar JA, et al. Survival following primary surgery for oral cancer. Oral Oncol. 2009;45:201.11. Rogers SN, Brown JS, Woolgar JA, et al. Survival following primary surgery for oral cancer. Oral Oncol. 2009;45:201.11.
6.
go back to reference Shaw RJ, Lowe D, Woolgar JA, et al. Extracapsular spread in oral squamous cell carcinoma. Head Neck. 2010;32:714.22. Shaw RJ, Lowe D, Woolgar JA, et al. Extracapsular spread in oral squamous cell carcinoma. Head Neck. 2010;32:714.22.
7.
go back to reference Brennan JA, Mao L, Hruban RH, et al. Molecular assessment of histopathological staging in squamous-cell carcinoma of the head and neck. N Engl J Med. 1995;332:429.35. Brennan JA, Mao L, Hruban RH, et al. Molecular assessment of histopathological staging in squamous-cell carcinoma of the head and neck. N Engl J Med. 1995;332:429.35.
8.
go back to reference Huang X, Pateromichelakis S, Hills A, et al. p53 mutations in deep tissues are more strongly associated with recurrence than mutation-positive mucosal margins. Clin Cancer Res. 2007;13:6099.106. Huang X, Pateromichelakis S, Hills A, et al. p53 mutations in deep tissues are more strongly associated with recurrence than mutation-positive mucosal margins. Clin Cancer Res. 2007;13:6099.106.
9.
go back to reference Graveland AP, Golusinski PJ, Buijze M, et al. Loss of heterozygosity at 9p and p53 immunopositivity in surgical margins predict local relapse in head and neck squamous cell carcinoma. Int J Cancer. 2011;128:1852.9. Graveland AP, Golusinski PJ, Buijze M, et al. Loss of heterozygosity at 9p and p53 immunopositivity in surgical margins predict local relapse in head and neck squamous cell carcinoma. Int J Cancer. 2011;128:1852.9.
10.
go back to reference Temam S, Casiraghi O, Lahaye JB, et al. Tetranucleotide microsatellite instability in surgical margins for prediction of local recurrence of head and neck squamous cell carcinoma. Clin Cancer Res. 2004;10(12 Pt 1):4022.8. Temam S, Casiraghi O, Lahaye JB, et al. Tetranucleotide microsatellite instability in surgical margins for prediction of local recurrence of head and neck squamous cell carcinoma. Clin Cancer Res. 2004;10(12 Pt 1):4022.8.
11.
go back to reference Shaw R. The epigenetics of oral cancer. Int J Oral Maxillofac Surg. 2006;35:101.8. Shaw R. The epigenetics of oral cancer. Int J Oral Maxillofac Surg. 2006;35:101.8.
12.
go back to reference Shaw RJ, Liloglou T, Rogers SN, et al. Promoter methylation of P16, RARbeta, E-cadherin, cyclin A1 and cytoglobin in oral cancer: quantitative evaluation using pyrosequencing. Br J Cancer. 2006;94:561.8. Shaw RJ, Liloglou T, Rogers SN, et al. Promoter methylation of P16, RARbeta, E-cadherin, cyclin A1 and cytoglobin in oral cancer: quantitative evaluation using pyrosequencing. Br J Cancer. 2006;94:561.8.
13.
go back to reference Eads CA, Danenberg KD, Kawakami K, et al. MethyLight: a high-throughput assay to measure DNA methylation. Nucleic Acids Res. 2000;28:E32.PubMedCrossRef Eads CA, Danenberg KD, Kawakami K, et al. MethyLight: a high-throughput assay to measure DNA methylation. Nucleic Acids Res. 2000;28:E32.PubMedCrossRef
14.
go back to reference Shaw RJ, Akufo-Tetteh EK, Risk JM, Field JK, Liloglou T. Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing. Nucleic Acids Res. 2006;34:e78.PubMedCrossRef Shaw RJ, Akufo-Tetteh EK, Risk JM, Field JK, Liloglou T. Methylation enrichment pyrosequencing: combining the specificity of MSP with validation by pyrosequencing. Nucleic Acids Res. 2006;34:e78.PubMedCrossRef
15.
go back to reference Goldenberg D, Harden S, Masayesva BG, et al. Intraoperative molecular margin analysis in head and neck cancer. Arch Otolaryngol Head Neck Surg. 2004;130:39.44. Goldenberg D, Harden S, Masayesva BG, et al. Intraoperative molecular margin analysis in head and neck cancer. Arch Otolaryngol Head Neck Surg. 2004;130:39.44.
16.
go back to reference Martone T, Gillio-Tos A, De Marco L, et al. Association between hypermethylated tumor and paired surgical margins in head and neck squamous cell carcinomas. Clin Cancer Res. 2007;13:5089.94. Martone T, Gillio-Tos A, De Marco L, et al. Association between hypermethylated tumor and paired surgical margins in head and neck squamous cell carcinomas. Clin Cancer Res. 2007;13:5089.94.
17.
go back to reference Shaw RJ, Hall GL, Woolgar JA, et al. Quantitative methylation analysis of resection margins and lymph nodes in oral squamous cell carcinoma. Br J Oral Maxillofac Surg. 2007;45:617.22. Shaw RJ, Hall GL, Woolgar JA, et al. Quantitative methylation analysis of resection margins and lymph nodes in oral squamous cell carcinoma. Br J Oral Maxillofac Surg. 2007;45:617.22.
18.
go back to reference Sinha P, Bahadur S, Thakar A, et al. Significance of promoter hypermethylation of p16 gene for margin assessment in carcinoma tongue. Head Neck. 2009;31:1423.30. Sinha P, Bahadur S, Thakar A, et al. Significance of promoter hypermethylation of p16 gene for margin assessment in carcinoma tongue. Head Neck. 2009;31:1423.30.
19.
go back to reference Tan HK, Saulnier P, Auperin A, et al. Quantitative methylation analyses of resection margins predict local recurrences and disease-specific deaths in patients with head and neck squamous cell carcinomas. Br J Cancer. 2008;99:357.63. Tan HK, Saulnier P, Auperin A, et al. Quantitative methylation analyses of resection margins predict local recurrences and disease-specific deaths in patients with head and neck squamous cell carcinomas. Br J Cancer. 2008;99:357.63.
20.
go back to reference Yang B, Gao YT, Du Z, Zhao L, Song WQ. Methylation-based molecular margin analysis in hepatocellular carcinoma. Biochem Biophys Res Commun. 2005;338:1353.8. Yang B, Gao YT, Du Z, Zhao L, Song WQ. Methylation-based molecular margin analysis in hepatocellular carcinoma. Biochem Biophys Res Commun. 2005;338:1353.8.
21.
go back to reference Guo M, House MG, Hooker C, et al. Promoter hypermethylation of resected bronchial margins: a field defect of changes? Clin Cancer Res. 2004;10:5131.6. Guo M, House MG, Hooker C, et al. Promoter hypermethylation of resected bronchial margins: a field defect of changes? Clin Cancer Res. 2004;10:5131.6.
22.
go back to reference Shaw RJ, Hall GL, Lowe D, et al. CpG island methylation phenotype (CIMP) in oral cancer: associated with a marked inflammatory response and less aggressive tumour biology. Oral Oncol. 2007;43:878.86. Shaw RJ, Hall GL, Lowe D, et al. CpG island methylation phenotype (CIMP) in oral cancer: associated with a marked inflammatory response and less aggressive tumour biology. Oral Oncol. 2007;43:878.86.
23.
go back to reference Schache AG, Hall G, Woolgar JA, et al. Quantitative promoter methylation differentiates carcinoma ex pleomorphic adenoma from pleomorphic salivary adenoma. Br J Cancer. 2010;103:1846.51. Schache AG, Hall G, Woolgar JA, et al. Quantitative promoter methylation differentiates carcinoma ex pleomorphic adenoma from pleomorphic salivary adenoma. Br J Cancer. 2010;103:1846.51.
24.
go back to reference Hall GL, Shaw RJ, Field EA, et al. p16 Promoter methylation is a potential predictor of malignant transformation in oral epithelial dysplasia. Cancer Epidemiol Biomarkers Prev. 2008;17:2174.9. Hall GL, Shaw RJ, Field EA, et al. p16 Promoter methylation is a potential predictor of malignant transformation in oral epithelial dysplasia. Cancer Epidemiol Biomarkers Prev. 2008;17:2174.9.
25.
go back to reference Brown JS, Blackburn TK, Woolgar JA, et al. A comparison of outcomes for patients with oral squamous cell carcinoma at intermediate risk of recurrence treated by surgery alone or with post-operative radiotherapy. Oral Oncol. 2007;43:764.73. Brown JS, Blackburn TK, Woolgar JA, et al. A comparison of outcomes for patients with oral squamous cell carcinoma at intermediate risk of recurrence treated by surgery alone or with post-operative radiotherapy. Oral Oncol. 2007;43:764.73.
Metadata
Title
Molecular Staging of Surgical Margins in Oral Squamous Cell Carcinoma Using Promoter Methylation of p16INK4A, Cytoglobin, E-cadherin, and TMEFF2
Authors
Richard J. Shaw, MD
Andrew J. Hobkirk, MBChB, BDS
George Nikolaidis, PhD
Julia A. Woolgar, PhD
Asterios Triantafyllou, PhD
James S. Brown, MD
Triantafillos Liloglou, PhD
Janet M. Risk, PhD
Publication date
01-08-2013
Publisher
Springer US
Published in
Annals of Surgical Oncology / Issue 8/2013
Print ISSN: 1068-9265
Electronic ISSN: 1534-4681
DOI
https://doi.org/10.1245/s10434-012-2713-8

Other articles of this Issue 8/2013

Annals of Surgical Oncology 8/2013 Go to the issue