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J Craniomaxillofac Surg. 2015 Oct;43(8):1494-500. doi: 10.1016/j.jcms.2015.07.028. Epub 2015 Aug 4.

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.

Author information

1
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.
2
Department of Biomedical and Neuro-Muscular Sciences, Section of Oral Science (Head: Prof. Lucio Montebugnoli, DDS), University of Bologna, Bologna, Italy.
3
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.
4
Department of Biomedical and Neuro-Muscular Sciences, Section of Maxillofacial Surgery (Head: Prof. Claudio Marchetti, MD), University of Bologna, Italy.

Abstract

PURPOSE:

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.

MATERIAL AND 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).

CONCLUSION:

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.

KEYWORDS:

DNA methylation; Next generation sequencing; Oral scraping; Oral squamous cell carcinoma

PMID:
26302938
DOI:
10.1016/j.jcms.2015.07.028
[Indexed for MEDLINE]

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