title: DNA methylation analysis by bisulfite next-generation sequencing for early detection of oral squamous cell carcinoma and high-grade squamous intraepithelial lesion from oral brushing.
abstract: OBJECTIVE
  Oral squamous cell carcinoma (OSCC) is commonly preceded by oral potentially malignant lesions (OPML). The aim of the present study was to assess, by bisulfite next-generation sequencing (NGS), the methylation status of a list of candidate genes obtained from oral brushings to early detect OPML and OSCC.
  
  
  METHODS
  Oral brushing specimens from 11 OSCC, 11 high-grade squamous intraepithelial lesions (HG-SIL), 9 low-grade SIL (LG-SIL), 9 oral lichen planus (OLP), and 8 healthy donors were included in this study. We investigated, by means of bisulfite NGS, the promoter of GP1BB, ZAP70, KIF1A, p16[CDKN2A], CDH1, miR137, and miR375. Statistical significance between lesions and a pool of healthy donors were evaluated with the Mann-Whitney U test.
  
  
  RESULTS
  ZAP70 was found to be hypermethylated in 100% of OSCC and HG-SIL and in 28.6% of LG-SIL. GP1BB hypomethylation was detected in 90.9% OSCC and HG-SIL and in 37.5% of LG-SIL. MiR137 was hypermethylated in 100% of OLP, 44.4% of OSCC, 40% HG-SIL, and 25% LG-SIL. KIF1A hypermethylation was found to be associated with TP53 mutations (p < 0.0001).
  
  
  CONCLUSIONS
  In the present preliminary cohort of patients, DNA methylation analysis of GP1BB and ZAP70 seems to be a promising noninvasive tool for early detection of OSCC and HG SIL from oral brushing specimens.
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Anatomic Pathology "M.Malpighi" at Bellaria Hospital (Head: Prof. Maria Pia Foschini, MD), University of Bologna, Italy.
authors: Luca
authors: L
authors: Morandi
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Oral Science (Head: Prof. Lucio Montebugnoli, DDS), University of Bologna, Bologna, Italy.
authors: Davide
authors: D
authors: Gissi
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Maxillofacial Surgery (Head: Prof. Claudio Marchetti, MD), University of Bologna, Italy. Electronic address: achille.tarsitano2@unibo.it.
authors: Achille
authors: A
authors: Tarsitano
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Anatomic Pathology "M.Malpighi" at Bellaria Hospital (Head: Prof. Maria Pia Foschini, MD), University of Bologna, Italy.
authors: Sofia
authors: S
authors: Asioli
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Anatomic Pathology "M.Malpighi" at Bellaria Hospital (Head: Prof. Maria Pia Foschini, MD), University of Bologna, Italy.
authors: Valentina
authors: V
authors: Monti
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Maxillofacial Surgery (Head: Prof. Claudio Marchetti, MD), University of Bologna, Italy.
authors: Giacomo
authors: G
authors: Del Corso
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Maxillofacial Surgery (Head: Prof. Claudio Marchetti, MD), University of Bologna, Italy.
authors: Claudio
authors: C
authors: Marchetti
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Oral Science (Head: Prof. Lucio Montebugnoli, DDS), University of Bologna, Bologna, Italy.
authors: Lucio
authors: L
authors: Montebugnoli
authors: Department of Biomedical and Neuro-Muscular Sciences, Section of Anatomic Pathology "M.Malpighi" at Bellaria Hospital (Head: Prof. Maria Pia Foschini, MD), University of Bologna, Italy.
authors: Maria Pia
authors: MP
authors: Foschini
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD; D015820:Cadherins / Q000235:genetics; D002278:Carcinoma in Situ / Q000175:diagnosis* / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000235:genetics; D015331:Cohort Studies; D018899:CpG Islands / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics; D003581:Cytodiagnosis / Q000379:methods*; D019175:DNA Methylation / Q000235:genetics; D055088:Early Detection of Cancer / Q000379:methods*; D005091:Exons / Q000235:genetics; D005260:Female; D006801:Humans; D016547:Kinesins / Q000235:genetics; D017676:Lichen Planus, Oral / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000235:genetics; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000175:diagnosis* / Q000235:genetics; D009154:Mutation / Q000235:genetics; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D019941:Cyclin-Dependent Kinase Inhibitor p16; C545567:MIRN137 microRNA, human; C540933:MIRN375 microRNA, human; D035683:MicroRNAs; D019038:Platelet Glycoprotein GPIb-IX Complex; D013447:Sulfites; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C094990:adhesion receptor; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human; D016547:Kinesins; C042345:hydrogen sulfite
keywords: DNA methylation; Next generation sequencing; Oral scraping; Oral squamous cell carcinoma