Objective: To explore the downregulation of ATP-binding cassette transporter B subfamily member 1 (ABCB1) by Tepoxalin mediated AKT/mTOR pathway in regulating the migration and apoptosis of colorectal cancer (CRC) LS1034 cells and its possible mechanisms.Methods: Public databases (GEPIA and HPA) were used to analyze the expression level and prognosis of ABCB1 in CRC, CCK-8 was used to detect cell proliferation, RT-qPCR and Western blot were used for mRNA and protein detection, flow cytometry was used for apoptosis detection, and Transwell assay was used to measure cell migration. Results: ABCB1 was highly expressed in CRC tissues (P0.05). Cell validation showed that ABCB1 was an upregulated gene in CRC. Tepoxalin inhibits ABCB1 in colorectal cancer cells and blocks the induction of proliferation, migration, and apoptosis in colorectal cancer cells. In addition, Western blot results showed that tepoxalin could inhibit AKT/mTOR signaling pathway and apoptotic signaling pathway expression.Conclusion: Tepoxalin inhibits proliferation and migration of CRC cells while inducing apoptosis by targeting ABCB1, which is mediated by ABCB1-mediated inhibition of AKT/mTOR signaling pathway.