Evaluation of NID2 promoter methylation for screening of Oral squamous cell carcinoma

Abstract

Background: Oral squamous cell carcinoma (OSCC) is an aggressive human malignancy. Because of late diagnosis and recurrence of OSCC, the treatment of patients with OSCC is often ineffective. Thus, finding novel biomarkers of OSCC are essential. Here we derived a methylation marker by utilizing methylation microarray data and testing its capacity in cross-sectional study designed for OSCC detection and screening. Methods: According to bioinformatics analysis of total of 27,578 cg sites, cg22881914 of Nidogen 2 (NID2) methylation was selected for evaluation. Next, we confirmed the methylation status by bisulfite sequencing from the microdissected OSCC cells in comparison with the microdissected oral epithelia. Subsequently, we developed a simple technique using real-time PCR with the specific probe to examine the ability for the detection of OSCC in the oral epithelial samples, which included 103 oral rinse and 82 oral swab samples. Results: Based on the comparison of microdissected tissue, cg22881914 of NID2 was proved to be methylated in most OSCC cells but unmethylated in the normal oral epithelia. Furthermore, the methylated NID2-relied quantitative PCR approach has demonstrated that this marker assists in distinguishing among patients with OSCC from normal oral epithelia, smokers, and patients with oral lichen planus using the non-invasive oral rinse and swab samples. Conclusions: Specific methylation at cg22881914 of NID2 of OSCC could be used as an important potential marker for detecting OSCC. Thus, to certify the utility of this marker, further studies with a larger sample size are needed.

Analysis of the Prognostic Value and Gene Expression Mechanism of SHOX2 in Lung Adenocarcinoma

“…Emerging molecular diagnostic methods are being increasingly applied in clinical practice because they are more sensitive and objective. DNA methylation alteration is considered an early predictor of cancer and can be detected during the early stages of tumorigenesis ( Zhang Y. et al, 2016 ; Dirks et al, 2016 ; Wang et al, 2018 ; Srisuttee et al, 2020 ). Because of the hypermethylation of SHOX2 in lung cancer, methylation detection has been applied to assist in the early diagnosis of unknown pulmonary nodules ( Kneip et al, 2011 ; Konecny et al, 2016 ; Weiss et al, 2017 ; Shi et al, 2020 ).…”

Salivary DNA Methylation as an Epigenetic Biomarker for Head and Neck Cancer. Part I: A Diagnostic Accuracy Meta-Analysis

“…After title and abstract review, 27 articles were submitted for full-text reading, of which nine were excluded for the following reasons: non-independent cancer group (two articles); reviews, letters, personal opinions, book chapters, case reports, conference abstracts, and meetings (three articles); absence of a healthy control group (one article); saliva enriched with brush oral cytology (two articles); and insufficient information for meta-analysis (one article). In the end, 18 articles met the inclusion criteria for final analysis [ 10 , 11 , 12 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 ].…”

“…A total of 34 different genes were identified in the studies. Five studies evaluated the methylation status of a single gene [ 31 , 34 , 36 , 39 , 41 ] and 13 studies evaluated two or more genes [ 10 , 11 , 12 , 30 , 32 , 33 , 35 , 37 , 38 , 40 , 42 , 43 , 44 ]. Ten studies focused on gene promoter methylation panels combining two to four genes, whereas eight studies evaluated only single genes.…”

Saliva Gene Promoter Hypermethylation as a Biomarker in Oral Cancer

“…High salivary methylation of NID2 was observed in oral cancer patients whereas no methylation was detected in smokers and healthy controls. In addition, sensitivity increased up to 90.91% when the NID2 methylation was detected in oral swabs from matched oral cancer patients (n = 22), which could be explained by the larger number of epithelial cells from cancer lesions using oral swabs vs. salivary oral rinses [ 41 ].…”