Peripheral blood-based DNA methylation panel for auxiliary diagnosis of esophageal squamous cell carcinoma

The less than 20% five-year survival of esophageal squamous cell carcinoma (ESCC) is mostly due to the lack of early diagnosis. We aimed to develop a blood-based DNA methylation profile for the auxiliary diagnosis of ESCC. DNA methylation levels were quantitatively determined by mass spectrometry. Logistic regression adjusted for covariates and receiver operating characteristic (ROC) curve were used for data analyses. Sixteen CpG sites in HYAL2 , S100P and DYRK4 were investigated in two independent case-control studies with a total of 601 ESCC cases and 802 healthy controls. In Study I, hypomethylation was found in 11 CpG sites of the three genes (odds ratios (ORs) per 10% decrease in methylation: 1.35–1.94, P -values < 0.004). In Study II with earlier cases, hypomethylation was validated in 9 CpG sites of HYAL2 and S100P (ORs per 10% decrease in methylation: 1.27–1.69, P -values < 0.001). In both studies, methylation differences between cases and controls were increased in advanced or more aggressive tumors. A methylation panel was established to distinguish ESCC cases from controls with area under the curve (AUC) values of 0.68–0.78 in Study I, and AUC values of 0.70–0.86 in Study II. When complete blood count (CBC) parameters were further adjusted for in the methylation panel of Study II, the discriminatory ability was markedly improved, yielding an AUC value of 0.80–0.91 for the detection of ESCC. With a large sample size, our study reported a peripheral blood-based panel using quantitative methylation signatures of HYAL2 , S100P and DYRK4 genes and CBC parameters, suggesting a feasible and efficient strategy for the in vitro auxiliary diagnosis of ESCC.