In this study, we also found downregulation of ZNF667-AS1 in esophageal cancer cell lines and ESCC tissues, and proximal promoter hypermethylation may be one of the mechanisms in leading to its silencing; together with the inhibiting effect of ZNF667-AS1 on esophageal cancer cells viability, migration, and invasion; suggesting the tumor-suppressor role of ZNF667-AS1 in ESCC tumorigenesis. Here, ZNF667 is linked to esophageal squamous cell carcinoma.