In addition to identifying the well-defined cancer-associated genes TP53, NOTCH1 (MIM 190198), PIK3CA, RB1, CDKN2A, and FBXW7, we identified recently implicated ESCC-associated genes AJUBA, ZNF750, and FAT1 in our combined cohort. Recently, pan-cancer studies across 21 cancer types predicted that AJUBA and ZNF750, two SMGs in our cohort, are associated with some types of cancer.50 Using functional studies, we found that AJUBA and ZNF750 (previously known but with unknown function in ESCC) act as tumor suppressors and that mutations in these genes abrogated their tumor-suppressive effects. Therefore, AJUBA and ZNF750 mutations encoding truncated or disrupted proteins might contribute to ESCC tumorigenesis.