Oral brushing as a useful method to study epigenetic alterations in oral mucosa from OSCC patients

A non invasive method for the early diagnosis in Oral Oncology may have a major impact on survival and quality of life. Aim of the present study was to evaluate the feasibility of a new non-invasive assay, based on brushing of oral mucosa and DNA methylation analysis using quantitative Bisulfite-Next Generation Sequencing. A total of 684 differentially methylated genes in OSCC was investigated by NGS, a set of 8 genes was tested in 48 patients and remaining 25 patients was tested for 12 genes. In each sample, fluorimeter quantus was used for a quantitative DNA evaluation, and DNA amount >1 ng/ul was considered adequate for DNA methylation analysis. PAP smear evaluation showed presence of keratinocytes of the upper and medium layer within the brush. Oral brushing seems to be a reliable procedure to obtain adequate DNA amounts from keratinocytes, and it may be used as a not invasive, easy to-perform method to study epigenetic alterations, such DNA methylation pattern, in oral mucosa.