Our methylation-array analysis identified 10 genes (HOXB2, NEFL, SLC15A3, OBSL1, MIR124-2, KDM2B, PAX6, PLEC, SEPT9, HOXD9) as candidates for predictive biomarkers of LNM in a discovery cohort of 67 ESCC cases. Here, HOXB2 is linked to esophageal squamous cell carcinoma.