In addition, univariate analysis indicated that the factors significantly associated with OS were Bin1 methylation status, TNM stage, invasion depth, tumor differentiation grade and lymph node metastasis (all P < 0.001), whereas age and gender were not related to the OS time of ESCC patients (P = 0.971; P = 0.994). This evidence concerns the gene BIN1 and metastatic malignant neoplasm in the lymph nodes.