Objective: The aim of this study was to investigate the regulatory effect of dagliflozin on the KLF4-NDRG2 signaling pathway in colon cancer cells, and to clarify the effect of dagliflozin on the process of colon cancer cell development. Methods: 10 colon cancer RKO cell strains were divided into control group and experimental group, the starting time of the study was July 2023, and the ending time was October 2023, the number of cells in each group was 5. The cells in the control group were applied to spread RKO cells into 6-well plates, and the cell strains in the experimental group were applied to pre-stimulation with 2 μm dagliflozin, and the improvement of the lineation of the two cell strains was observed by taking the same field of view, while the migration distance of the cells in the control group was calculated and compared with that of RKO cells. At the same time, the cell migration distance was calculated to compare the in vitro motility of RKO cells, and the KLF4 mRNA and protein expression levels of the two cell strains were detected by RT-qPCR. At the same time, the KLF4 mRNA and protein expression levels of the two cell strains were examined by RT-qPCR. RESULTS: The levels of strain KLF4 mRNA and protein expression of RKO cells in the experimental group were lower than those in the control group after dagliflozin intervention, and the differences were statistically significant (P<0.05). And the in vitro motility of strain RKO cells in the experimental group was significantly lower than that of the control group, and the difference was statistically significant (P<0.05). Conclusion: Dagliflozin can play an inhibitory role on the invasive ability of RKO cells, and also effectively reduce the expression level of KLF4, which can have an important impact on colon cancer occurrence and progression by regulating the KLF4-NDRG2 signaling pathway.