In our present study, we found that PBK (area under the curve [AUC]= 96.5%), KIF2C (AUC= 98.8%), NUF2 (AUC= 99.2%), KIF20A (AUC= 99.2%), RAD51AP1 (AUC= 96.7%), and DEPDC1 (AUC= 98.6%) all with a high sensitivity and specificity for the diagnosis of ESCC. This evidence concerns the gene PBK and esophageal squamous cell carcinoma.