KIF1A and EDNRB are differentially methylated in primary HNSCC and salivary rinses

Abstract

Silencing of tumor suppressor genes plays a vital role in head and neck carcinogenesis. In this study, we aimed to evaluate to the utility of aberrant promoter hypermethylation for detection in a panel of 10 genes (KIF1A, EDNRB, CDH4, TERT, CD44, NISCH, PAK3, VGF, MAL and FKBP4) in head and neck squamous cell carcinoma (HNSCC) via a candidate gene approach. We investigated methylation of the gene promoters by bisulfite modification and quantitative methylation-specific PCR (Q-MSP) in a preliminary study of a limited cohort of salivary rinses from healthy subjects (n 5 61) and patients with HNSCC (n 5 33). The methylation status of 2 selected genes (EDNRB and KIF1A) were then analyzed in 15 normal mucosa samples from a healthy population, 101 HNSCC tumors and the corresponding salivary rinses from 71 out of the 101 HNSCC patients were collected before treatment. The promoter regions of CDH4, TERT, VGF, MAL, FKBP4, NISCH and PAK3 were methylated in normal salivary rinses while no methylation of CD44 was observed in either normal salivary rinses or tumor samples. However, KIF1A and EDNRB were methylated in 98 and 97% of primary HNSCC tissues respectively and were only methylated in 2 and 6.6% of normal salivary rinses. In addition, KIF1A and EDNRB were methylated in 38 and 67.6% of salivary rinses from HNSCC patients, respectively. Promoter hypermethylation of KIF1A and EDNRB is a frequent event in primary HNSCC, and these genes are preferentially methylated in salivary rinses from HNSCC patients. KIF1A and EDNRB are potential biomarkers for HNSCC detection.Among human malignancies, head and neck cancer is the sixth most common cancer in the world. 1 More than 40,000 new cases of head and neck squamous cell carcinoma (HNSCC) are diagnosed in the United States each year, with a mortality rate of 12,000 U.S. deaths annually. Survival rates have not improved significantly for patients with HNSCC in the past 30 years despite active clinical and basic science research addressing this issue. Molecular detection of HNSCC in body fluids has the potential to improve post-treatment surveillance, provide prognostic information, and influence therapy. Body fluids can potentially carry whole cells as well as protein, DNA and RNA species that allow for detection of cellular alterations related to cancer. In previous studies, body fluids such as sputum for lung cancer, 2 urine for urologic tumors, 3 salivary rinses for HNSCC, 4-7 and breast fluid for breast cancer 8 have been used in multiple detection strategies. [9][10][11][12][13] Silencing of tumor suppressor genes by means of promoter hypermethylation plays a role in head and neck carcinogenesis. 5 Measuring promoter hypermethylation by using real time quantitative methylation-specific PCR (Q-MSP) allows an objective, robust, and rapid assessment of promoter methylation status. The ability to quantify methylation

Correlation of gene methylation in surgical margin imprints with locoregional recurrence in head and neck squamous cell carcinoma

“…Frequent promoter methylation of EDNRB has been detected in gastric cancer, prostate cancer, and head and neck cancer 27, 37-39 . A study from our laboratory indicated this gene was methylated in 97% of primary HNSCC tissues and 6.6% of normal control salivary rinses 21 . Besides extremely high sensitivity and specificity, the comparatively high relative QMSP values in HNSCC combine to make these excellent tumor-specific markers for margin analysis.…”

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

“…As demonstrated in the literature, the DNA samples extracted from salivary rinses showed enough quality to carry out qMSP reactions [29,30,33,34].…”

Key tumor suppressor genes inactivated by “greater promoter” methylation and somatic mutations in head and neck cancer

“…Epigenetic events linked to TSG inactivation through promoter methylation, are more frequent events than somatic mutations in cancer, and may be driving tumorigenic initiation and progression. Promoter methylation of CDK2NA , HOXA9 , NID2 , EDNRB , KIF1A, and DCC have previously been identified and characterized in HNSCC 11 - 13 …”