To investigate the effect of escin on the proliferation，migration，and invasion of colorectal cancer cell line HCT116 induced by trimethylamine N-oxide(TMAO) and to explore its mechanism of action.Methods:HCT116 colorectal cancer cells were divided into a control group，a TMAO group，and an experimental group.The control group received no treatment，the TMAO group was induced with 100 μmol/L TMAO，and the experimental group was treated with 100 μmol/L TMAO followed by 10 μmol/L escin.Cell proliferation，migration，and invasion were assessed using the MTT assay，scratch wound healing assay，and Transwell migration and invasion assays，respectively.The expression levels of SRC，pBTK，and pTRIO proteins were measured by Western blotting.Results:TMAO induction enhanced the proliferation，migration，and invasion of HCT116 cells and significantly upregulated the mRNA levels of genes in the SRC-BTK-TRIO pathway.Escin treatment inhibited cancer cell proliferation(P<0.01)，reduced their migration(P<0.001) and invasion(P<0.01) capabilities，and downregulated the expression of SRC(P<0.05)，BTK(P<0.01)，and TRIO(P<0.05) proteins.Conclusion:Escin can significantly inhibit the proliferation，migration，and invasion of HCT116 cells by downregulating the SRC-BTK-TRIO pathway.