OBJECTIVES: The study examined the NDUFA1 expression and its prognostic value in pan-cancer, especially in esophageal cancer (ESCA). Its carcinogenic effect on ESCA was further elucidated. MATERIALS AND METHODS: TCGA database was used to examine NDUFA1 expression and its prognostic value in 33 cancer types. GO and KEGG were performed for function and pathway enrichment of NDUFA1-related genes. The carcinogenic effect on ESCA was verified in both KYSE-30 cell and Xenograft mouse model. RESULTS: Abnormal high expression of NDUFA1 was detected in pan-cancer, and related to immune cell infiltration. TCGA database indicated an elevated value of NDUFA1 in ESCA tumor tissues, which was linked to patients' poor prognosis. NDUFA1-related genes were mainly enriched in oxidative stress and immune response in ESCA. NDUFA1 knockdown significantly suppressed ESCA cell proliferation, migration and invasion. Similarly, tumor growth of ESCA xenograft mice was inhibited by NDUFA1 knockdown. Activated PI3K-Akt signaling was detected in both ESCA cell lines and tumor tissues, which was reversed by NDUFA1 knockdown. CONCLUSION: Multi-omics analysis showed that NDUFA1 might be adopted as a potential therapeutic goal and prognostic indicator for a variety of cancers, especially for ESCA. NDUFA1 knockdown inhibited ESCA tumor growth, which may have the participation of the PI3K/Akt signaling pathway.