Objective: To investigate the effect of Fascin shRNA on the grow th of colorectal cancer cells. Methods: Colorectal cancer cells HT29 infected with Fascin shRNA and shRNA control lentivirus, real-time PCR and Western blot were used to measure the expression of Fascin mRNA and protein, at the same time, the level of Notch1 protein was detected by Western blot, Notch1 signaling pathway inhibitor DAPT was used to the HT29 cells after downregulate Fascin. Cell proliferation, cell cycle, the expression levels of cyclin p27 and Cyclin D1 were detected by MTT assay, flow cytometry, and Western blot, respectively. Results: After Fascin shRNA lentivirus infection, Fascin transcription and expression levels were reduced. However, shRNA control lentivirus had no effect on Fascin expression. The cell proliferation ability of silent Fascin was reduced, the proportion of G₀/G₁ in cells increased, the level of Cyclin D1 protein in the cells decreased, the level of p27 protein was increased. Silencing Fascin reduced the level of Notch1 protein in cells. Moreover, Notch1 signaling pathway inhibitor DAPT coexisted with silencing Fascin to further inhibit the proliferation of colorectal cancer cells, the cell cycle of colorectal cancer was blocked, the expression of Cyclin D1 protein reduced, the expression of p27 protein promoted. Conclusion: Fascin shRNA can inhibit proliferation and blocked cell cycle of colorectal cancer cells by inhibiting Notch1 signaling pathway.