Background Salivary rinses have been recently proposed as a valuable resource for the development of epigenetic biomarkers for detection and monitoring of head and neck squamous cell carcinoma (HNSCC). Both salivary rinses collected with and without an exfoliating brush from patients with HNSCC are used in detection of promoter hypermethylation, yet their correlation of promoter hypermethylation has not been evaluated. This study was to evaluate the concordance of promoter hypermethylation between salivary rinses collected with and without an exfoliating brush from patients with HNSCC. Methodolgy 57 paired salivary rinses collected with or without an exfoliating brush from identical HNSCC patients were evaluated for promoter hypermethylation status using Quantitative Methylation-Specific PCR. Target tumor suppressor gene promoter regions were selected based on our previous studies describing a panel for HNSCC screening and surveillance, including P16, CCNA1, DCC, TIMP3, MGMT, DAPK and MINT31. Principal Findings In salivary rinses collected with and without brush, frequent methylation was detected in P16 (8.8% vs. 5.2%), CCNA1 (26.3% vs. 22.8%), DCC (33.3% vs. 29.8%), TIMP3 (31.6% vs. 36.8%), MGMT (29.8% vs. 38.6%), DAPK (14.0% vs. 19.2%), and MINT31 (10.5% vs. 8.8%). Spearman's rank correlation coefficient showed a positive correlation between salivary rinses collected with and without brush for P16 (ρ = 0.79), CCNA1 (ρ = 0.61), DCC (ρ = 0.58), TIMP3 (ρ = 0.10), MGMT (ρ = 0.70), DAPK (ρ = 0.51) and MINT31 (ρ = 0.72) (P<0.01). The percent agreement of promoter methylation between salivary rinses with brush and without brush were 96.5% for P16, 82.5% for CCNA1, 78.9% for DCC, 59.7% for TIMP3, 84.2% for MGMT, 84.2% for DAPK, and 94.7% for MINT31. Conclusions Our study demonstrated strong correlations of gene promoter hypermethylation between salivary rinses collected with and without an exfoliating brush. Salivary rinse collection without using an exfoliating brush may offer a cost effective, rapid, non-invasive, and reliable means for development of epigenetic salivary rinse biomarkers.
References
[1]
Haddad RI, Shin DM (2008) Recent advances in head and neck cancer. N Engl J Med 359: 1143–1154.
[2]
Leemans CR, Braakhuis BJ, Brakenhoff RH (2011) The molecular biology of head and neck cancer. Nat Rev Cancer 11: 9–22.
[3]
Franzmann EJ, Reategui EP, Pedroso F, Pernas FG, Karakullukcu BM, et al. (2007) Soluble CD44 is a potential marker for the early detection of head and neck cancer. Cancer Epidemiol Biomarkers Prev 16: 1348–1355.
[4]
Hu S, Arellano M, Boontheung P, Wang J, Zhou H, et al. (2008) Salivary proteomics for oral cancer biomarker discovery. Clin Cancer Res 14: 6246–6252.
[5]
Nagler R, Bahar G, Shpitzer T, Feinmesser R (2006) Concomitant analysis of salivary tumor markers–a new diagnostic tool for oral cancer. Clin Cancer Res 12: 3979–3984.
[6]
Tavassoli M, Brunel N, Maher R, Johnson NW, Soussi T (1998) p53 antibodies in the saliva of patients with squamous cell carcinoma of the oral cavity. Int J Cancer 78: 390–391.
[7]
Xie H, Onsongo G, Popko J, de Jong EP, Cao J, et al. (2008) Proteomics analysis of cells in whole saliva from oral cancer patients via value-added three-dimensional peptide fractionation and tandem mass spectrometry. Mol Cell Proteomics 7: 486–498.
[8]
Brinkmann O, Kastratovic DA, Dimitrijevic MV, Konstantinovic VS, Jelovac DB, et al. (2011) Oral squamous cell carcinoma detection by salivary biomarkers in a Serbian population. Oral Oncol 47: 51–55.
[9]
Li Y, Zhou X, St John MA, Wong DT (2004) RNA profiling of cell-free saliva using microarray technology. J Dent Res 83: 199–203.
[10]
Park NJ, Zhou H, Elashoff D, Henson BS, Kastratovic DA, et al. (2009) Salivary microRNA: discovery, characterization, and clinical utility for oral cancer detection. Clin Cancer Res 15: 5473–5477.
[11]
Ha PK, Califano JA (2006) Promoter methylation and inactivation of tumour-suppressor genes in oral squamous-cell carcinoma. Lancet Oncol 7: 77–82.
[12]
Demokan S, Chang X, Chuang A, Mydlarz WK, Kaur J, et al. (2010) KIF1A and EDNRB are differentially methylated in primary HNSCC and salivary rinses. Int J Cancer 127: 2351–2359.
[13]
Guerrero-Preston R, Soudry E, Acero J, Orera M, Moreno-Lopez L, et al. (2011) NID2 and HOXA9 Promoter Hypermethylation as Biomarkers for Prevention and Early Detection in Oral Cavity Squamous Cell Carcinoma Tissues and Saliva. Cancer Prev Res (Phila) 4: 1061–1072.
[14]
Kaur J, Demokan S, Tripathi SC, Macha MA, Begum S, et al. (2010) Promoter hypermethylation in Indian primary oral squamous cell carcinoma. Int J Cancer 127: 2367–2373.
[15]
Pattani KM, Califano J (2011) Long-Term Experience in Sentinel Node Biopsy for Early Oral and Oropharyngeal Squamous Cell Carcinoma. Ann Surg Oncol 18: 2709–2710.
[16]
Carvalho AL, Jeronimo C, Kim MM, Henrique R, Zhang Z, et al. (2008) Evaluation of promoter hypermethylation detection in body fluids as a screening/diagnosis tool for head and neck squamous cell carcinoma. Clin Cancer Res 14: 97–107.
[17]
Carvalho AL, Henrique R, Jeronimo C, Nayak C, Reddy A, et al. (2011) Detection of promoter hypermethylation in salivary rinses as a biomarker for head and neck squamous cell carcinoma surveillance. Clin Cancer Res 17: 4782–4789.
[18]
Rosas SL, Koch W, da Costa Carvalho MG, Wu L, Califano J, et al. (2001) Promoter hypermethylation patterns of p16, O6-methylguanine-DNA-methyltransferase, and death-associated protein kinase in tumors and saliva of head and neck cancer patients. Cancer Res 61: 939–942.
[19]
Spafford MF, Koch WM, Reed AL, Califano JA, Xu LH, et al. (2001) Detection of head and neck squamous cell carcinoma among exfoliated oral mucosal cells by microsatellite analysis. Clin Cancer Res 7: 607–612.
[20]
Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB (1996) Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A 93: 9821–9826.
[21]
Harden SV, Tokumaru Y, Westra WH, Goodman S, Ahrendt SA, et al. (2003) Gene promoter hypermethylation in tumors and lymph nodes of stage I lung cancer patients. Clin Cancer Res 9: 1370–1375.
[22]
Begum S, Brait M, Dasgupta S, Ostrow KL, Zahurak M, et al. (2011) An epigenetic marker panel for detection of lung cancer using cell-free serum DNA. Clin Cancer Res 17: 4494–4503.
[23]
Zhao M, Rosenbaum E, Carvalho AL, Koch W, Jiang W, et al. (2005) Feasibility of quantitative PCR-based saliva rinse screening of HPV for head and neck cancer. Int J Cancer 117: 605–610.
[24]
Kvalseth TO (1991) A Coefficient of Agreement for Nominal Scales - an Asymmetric Version of Kappa. Educational and Psychological Measurement 51: 95–101.
[25]
Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33: 159–174.
[26]
Sun W, Zaboli D, Wang H, Liu Y, Arnaoutakis D, et al. (2012) Detection of TIMP3 Promoter Hypermethylation in Salivary Rinse as an Independent Predictor of Local Recurrence-Free Survival in Head and Neck Cancer. Clin Cancer Res 18: 1082–91.
[27]
Lingen MW (2010) Screening for oral premalignancy and cancer: what platform and which biomarkers? Cancer Prev Res (Phila) 3: 1056–1059.
