Collectively, this study presented comprehensive proteomic characterization of EESCC and EDAC at multi-omics level, and revealed the functional pathways of cell cycle in EESCC, the positive impacts of apoptosis, and metabolic processes in EDAC, and further demonstrated the potential co-functions of RB1 and EGFR in ESCC, and the key role of MTOR at the protein and phosphoprotein levels in EDAC. The gene discussed is RB1; the disease is esophageal squamous cell carcinoma.