Background and Purpose: Esophageal squamous cell carcinoma (ESCC) is one of the most common malignancies and causes cancer-related deaths globally, necessitating the identification of novel therapeutic targets and treatments. Sterol O-acyltransferase 1 (SOAT1) is one of key genes related to the cholesterol esterification and identified as a singularly up-expression gene in ESCC tissues. However, the anti-cancer effects and molecular mechanisms of SOAT1 on ESCC remains unclear. Experimental Approach: Therapeutic target was screened and identified using ESCC patients’ samples and bioinformatics. To evaluate the anti-cancer effect of SOAT1 on ESCC, multiple experiences were performed such as SOAT1 knockdown, molecular docking analysis, CCK-8, clone formation assays, bioinformatics analysis, Western blotting and real-time PCR arrays. Key Results: Knockdown of SOAT1 could inhibit cell proliferation, migration, and invasion. Then, molecular docking analysis was used to identify piperine as a potential inhibitor of SOAT1. Interestingly, the inhibition of SOAT1 obviously enhanced ferroptosis and the inhibitory of cell viability. Furthermore, the inhibition of SOAT1 synergistically enhanced the anti-cancer effect of cisplatin in ESCC. Further information mining revealed that the inhibition of SOAT1 enhanced ALF4-induced ALOX12B expression by reducing PI3K-AKT pathway. Next, we found that SOAT1 inhibitor piperine suppressed tumor growth without affecting mice weight in vivo. Conclusion and Implications: Together, this study indicated that the inhibition of SOAT1 enhanced ferroptosis by reducing PI3K-AKT pathway and subsequently up-regulating ALF4/ALOX12B. Thus, our findings strongly emphasize that SOAT1 holds great potential as a highly promising clinical therapeutic target in ESCC.