Abstract: The main modalities for gastric cancer screening are limited to upper gastrointestinal\nendoscopy and contrast radiography. The former is invasive, and the latter has high false-negative\nrates. Thus, alternative diagnostic strategies are required. One solution may be a liquid biopsy.\nMethylated RUNX3 is a well-known biomarker of gastric cancer but it is very difficult to detect with\nconventional bisulfite-based methylation assays when only a small amount of serum is available. We\ndeveloped the combined restriction digital PCR (CORD) assay, a new methylation assay allowing\nfor the counting of as little as one copy of a methylated gene in a small sample of DNA without\nnecessitating DNA bisulfite treatment. We evaluated the sensitivity and specificity of the serum DNA\ntesting of methylated RUNX3 by the CORD assay for the detection of early gastric cancer using 50\npatients with early gastric cancer and 61 control individuals. The CORD assay had a sensitivity of\n50.0% and a specificity of 80.3% for early gastric cancer. Methylated RUNX3 copies were significantly\nassociated with tumor size, massive submucosal invasion, and lymph-vascular invasion. After the\ntreatment, the median number of methylated RUNX3 copies was significantly decreased. The CORD\nassay may provide an alternative screening strategy to detect even early-stage gastric cancer.
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