13-gene DNA Methylation Analysis from Oral Brushing: A Promising Non Invasive Tool in the Follow-up of Oral Cancer Patients

Abstract

Background: This study aimed to evaluate the prognostic value of a non-invasive sampling procedure based on 13-gene DNA methylation analysis in the follow-up of patients previously treated for oral squamous cell carcinoma (OSCC). Methods: The study population included 49 consecutive patients treated for OSCC. Oral brushing sample collection was performed at two different times: before any cancer treatment in the tumor mass and during patient follow-up almost 6 months after OSCC treatment, within the regenerative area after OSCC resection. Each sample was considered positive or negative in relation to a predefined cut-off value. Results: Before any cancer treatment, 47/49 specimens exceeded the score and were considered as positive. Six months after OSCC resection, 16/49 specimens also had positive scores in the samples collected from the regenerative area. During the follow-up period, 7/49 patients developed locoregional relapse: 6/7 patients had a positive score in the regenerative area after OSCC resection. The presence of a positive score after oral cancer treatment was the most powerful variable related to the appearance of locoregional relapse. Conclusion: 13-gene DNA methylation analysis by oral brushing may have a clinical application as a prognostic non-invasive tool in the follow-up of patients surgically treated for OSCC.

Droplet Digital PCR Analysis of Liquid Biopsy Samples Unveils the Diagnostic Role of hsa-miR-133a-3p and hsa-miR-375-3p in Oral Cancer

“…Indeed, it was widely demonstrated that epigenetic modifications represent early events of neoplastic transformation. In this context, the analysis of DNA methylation and miRNAs expression has been proposed for the early diagnosis of different tumors [52][53][54][55]. In line with the results achieved in this study, other studies demonstrated the diagnostic potential of both hsa-miR-133a-3p and hsa-miR-375-3p in different tumors, including gastric cancer, prostate cancer, hepatocellular carcinoma, colorectal cancer, etc., demonstrating the involvement of these miRNAs in cancer development, including that of oral cancer [56][57][58][59][60].…”

Clinical validation of 13‐gene DNA methylation analysis in oral brushing samples for detection of oral carcinoma: Italian multicenter study

“…In total, 96.6% ( n = 28) of OSCC and 100% ( n = 6) of high‐grade squamous intraepithelial lesion (HGSIL) specimens exceeded the threshold value, whereas none of the healthy donor samples ( n = 65) exceeded the threshold 13 . The clinical relevance of the method in oral oncology has been confirmed in patients surgically treated for OSCC; the 13‐gene DNA methylation analysis of samples taken from the regenerative area after OSCC resection could identify patients at risk of recurrence during follow‐up 14 …”

An Evolutionary Cancer Epigenetic Approach Revealed DNA Hypermethylation of Ultra-Conserved Non-Coding Elements in Squamous Cell Carcinoma of Different Mammalian Species

“…Oral brushing sample collection was performed in the population study as previously described [ 18 , 19 ]; in brief, a cytobrush (manufactured by N.H.M.P. Co., Ltd. PRC EC REP: Shanghai International trading corporation, Hamburg, Germany) was used to collect exfoliated cells from oral mucosa.…”

“…DNA methylation analysis was evaluated as previously described [ 18 , 19 ]. In brief, DNA from exfoliating brush specimens was purified using the MasterPure™ Complete DNA Purification Kit (Lucigen, Middleton, WI, USA, cod.…”