title: DNA methylation profiles and biomarkers of oral squamous cell carcinoma.
abstract: Oral squamous cell carcinoma (OSCC) constitutes >90% of oral cancers and is the sixth most common malignancy among males worldwide and the fourth leading cause of death due to cancer among males in Taiwan. However, most patients do not receive a diagnosis of OSCC until the late stages, which have a lower survival rate. The use of molecular marker analysis to identify early-stage OSCC would permit optimal timing for treatments and consequently prolong survival. The aim of this study was to identify biomarkers of OSCC using the Illumina GoldenGate Methylation Cancer Panel, which comprised a total of 1,505 CpG sites covering 807 genes. Samples of buccal mucosa resected from 40 OSCC patients and normal tissue samples obtained from 15 patients (normal mucosa from OSCC patients or from patients undergoing surgery unrelated to OSCC) were analyzed. Fms-related tyrosine kinase 4 (FLT4) methylation exhibited a perfect specificity for detecting OSCC, with an area under the receiver operating characteristic curve of 0.91 for both all-stage and early-stage OSCC. Methylation of 7 genes (ASCL1, FGF3, FLT4, GAS7, KDR, TERT, and TFPI2) constitutes the top-20 panels for detecting OSCC. The top-20 panels for detecting early-stage OSCC contain 8 genes: ADCYAP1, EPHA7, FLT4, GSTM2, KDR, MT1A, NPY, and TFPI2. FLT4 RNA expression and methylation level were validated using RT-PCR and a pyrosequencing methylation assay. The median level of FLT4 expression was 2.14-fold for normal relative to OSCC tissue samples (P < 0.0001). Among the 8 pyrosequenced FLT4 CpG sites, methylation level was much higher in the OSCC samples. In conclusion, methylation statuses of selected genes, and especially FLT4, KDR, and TFPI2, might be of great potential as biomarkers for early detection of buccal OSCC.
authors.affiliation: a Institute of Biostatistics; China Medical University ; Taichung , Taiwan.
authors.forename: Yu-Fen
authors.initials: YF
authors.lastname: Li
authors.forename: Yi-Hsiu
authors.initials: YH
authors.lastname: Hsiao
authors.forename: Yi-Hui
authors.initials: YH
authors.lastname: Lai
authors.forename: Yi-Chen
authors.initials: YC
authors.lastname: Chen
authors.forename: Ying-Ju
authors.initials: YJ
authors.lastname: Chen
authors.forename: Jian-Liang
authors.initials: JL
authors.lastname: Chou
authors.forename: Michael W Y
authors.initials: MW
authors.lastname: Chan
authors.forename: Yu-Hsing
authors.initials: YH
authors.lastname: Lin
authors.forename: Yung-An
authors.initials: YA
authors.lastname: Tsou
authors.forename: Ming-Hsui
authors.initials: MH
authors.lastname: Tsai
authors.forename: Chien-Kuo
authors.initials: CK
authors.lastname: Tai
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D018899:CpG Islands; D019175:DNA Methylation; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D006023:Glycoproteins; C021240:tissue-factor-pathway inhibitor 2; C581309:FLT4 protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3
keywords: DNA methylation array; FLT4; KDR; TFPI2; biomarkers; oral squamous cell carcinoma; pyrosequencing