Hypermethylation biomarkers for detection of head and neck squamous cell cancer

摘要

Differentially methylated oral squamous cell carcinoma (OSCC) biomarkers, identified in-vitro and validated in well-characterized surgical specimens, have shown poor clinical correlation in cohorts with different risk profiles. To overcome this lack of relevance we used the HumanMethylation27 BeadChip, publicly available methylation and expression array data, and Quantitative Methylation Specific PCR to uncover differential methylation in OSCC clinical samples with heterogeneous risk profiles. A two stage-design consisting of Discovery and Prevalence screens was used to identify differential promoter methylation and deregulated pathways in patients diagnosed with OSCC and head and neck squamous cell carcinoma. This Phase I Biomarker Development Trial identified a panel of differentially methylated genes in normal and OSCC clinical samples from patients with heterogeneous risk profiles. This panel may be useful for early detection and cancer prevention studies.

主权项

1. A method for identifying oral squamous cell cancer (OSCC) in a human patient comprising:
a) obtaining nucleic acid from a test sample taken from the oral cavity mucosal tissues of the patient, wherein the test sample is from a specimen selected from the group consisting of a tissue specimen, a biopsy specimen, a surgical specimen, saliva, and a cytological specimen; b) obtaining nucleic acid from an oral mucosal non-neoplastic reference sample wherein the reference sample is from a specimen selected from the group consisting of a tissue specimen, a biopsy specimen, a surgical specimen, saliva, and a cytological specimen; c) performing bisulfite modification to the nucleic acid in steps a) and b); d) performing quantitative methylation specific PCR (QMSP) on the bisulfite modified nucleic acid from step c) using PCR primers and probes specific for the promoter region of the HOXA9 and NID2 genes, wherein the primers and probes are SEQ ID NOS:10-15; e) determining the promoter methylation level of the promoter regions HOXA9 and NID2 in the test sample and the reference sample; f) comparing the level of promoter methylation of HOXA9 and NID2 from the test sample of the patient, to the level of promoter methylation of HOXA9 and NID2 in the reference sample; g) identifying said patient as having OSCC when the level of methylation of the promoter regions of HOXA9 and NID2 are increased relative to the level of methylation of the promoter regions of HOXA9 and NID2 in the reference sample.