EDNRB and DCC Salivary Rinse Hypermethylation Has a Similar Performance as Expert Clinical Examination in Discrimination of Oral Cancer/Dysplasia versus Benign Lesions

Abstract

Purpose Promoter hypermethylation has been recently proposed as a mean for HNSCC detection in salivary rinses. In a prospective study of a high-risk population, we showed that EDNRB promoter methylation in salivary rinses is a useful biomarker for oral cancer and premalignancy. Experimental Design Using that cohort, we evaluated EDNRB methylation status and 8 additional genes. Clinical risk assessment by expert clinicians was performed and compared with biomarker performance in the prediction of premalignant and malignant disease. Methylation status of 9 genes was analyzed in salivary rinses of 191 patients by Quantitative methylation-specific PCR. Results HOXA9, EDNRB and DCC methylation were associated (p= 0.012; p<0.0001; p=0.0005) with premalignant or malignant disease. On multivariable modeling, histological diagnosis was only independently associated with EDNRB (p=0.0003) or DCC (p=0.004) methylation. A subset of patients received clinical risk classification (CRC) by expert clinicians based on lesion examination. CRC, DCC and EDNRB were associated with diagnosis of dysplasia/cancer on univariate (p=0.008; p=0.026; p=0.046) and multivariate analysis (p=0.012; p=0.037; p=0.047). CRC identified dysplasia/cancer with56% of sensitivity and 66% of specificity with a similar AUC (0.61, 95%CI=0.60-0.81) when compared to EDNRB and DCC combined AUC (0.60, 95%CI=0.51-0.69), sensitivity of 46% and specificity of 72%. A combination of EDNRB, DCC and CRC was optimal AUC (0.67, 95%CI=0.58-0.76). Conclusion EDNRB and/or DCC methylation in salivary rinses compares well to examination by an expert clinician in CRC of oral lesions. These salivary biomarkers may be particularly useful in oral premalignancy and malignancy screening in clinical care settings in which expert clinicians are not available

Clinical implications of epigenetic regulation in oral cancer

“…Our pooled meta-analysis of 2194 oral cancer patients and 456 controls, demonstrated p16 hypermethylation in 43% oral cancers and 5.4% normal samples (Table 1). Several studies demonstrated that normal individuals with no tobacco habits showed absence or low levels of hypermethylation [9,17,22,24]. Our studies in tobacco chewing Indian oral cancer patients demonstrated p16 hypermethylation in 34% long term tobacco users (LTTUs), 25% premalignant tissues, 65% oral cancers, 47% tumor adjacent mucosa (TAM), and 0% normal controls with no tobacco habits [9].…”

“…Fonseca-Silva and coworkers demonstrated p16 promoter methylation in 78% HPV-16 positive oral cancers. Hypermethylation of p16 is observed in various sites of oral cavity including tongue, lower gingiva and oral floor [8,22,[25][26][27]. Our pooled meta-analysis of 2194 oral cancer patients and 456 controls, demonstrated p16 hypermethylation in 43% oral cancers and 5.4% normal samples (Table 1).…”

“…DNA methylation is detected in adjacent normal tissues indicating the occurrence as an 'early event' and/ or process of 'field cancerization' in oral cavity [9,21]. Studies by Califano and coworkers demonstrated promoter hypermethylation in endothelin receptor type B (EDNRB) and deleted in colon cancer (DCC) genes in dysplasia and cancer tissues using salivary rinses, thus providing a non-invasive biomarker in dysplastic tissues and oral cancer [22]. Development of a convenient tool for detecting early events in oral carcinogenesis may result in down staging of the cancer, early treatment, better recovery and survival of patients.…”

Expression and epigenetic regulation of DACT1 and DACT2 in oral squamous cell carcinoma

“…Recent studies in the literature validate the use of salivary rinse for biomarker detection [29][30][31][32]. As demonstrated in the literature, the DNA samples extracted from salivary rinses showed enough quality to carry out qMSP reactions [29,30,33,34].…”

Aberrant methylated EDNRB can act as a potential diagnostic biomarker in sporadic colorectal cancer while KISS1 is controversial

“…13,16,17 The interaction of this receptor with endothelins leads to the releasing of InsP 3 and Ca 2C as second messengers. 18 It is suggested that EDNRB gene is one of the important causes of Hirschsprung's disease (HSCR). 17 In the tumor development, transcript expression of this gene is decreased through hypermethylation of CpG-rich promoter regions resulting in alteration of ET1 signaling pathway, 19 which leads into proliferation, angiogenesis, and metastasis of tumors.…”