ObjectiveTo investigate the anti-liver cancer mechanism of water extract of Jianpi Xiaoji prescription and its effect on cell autophagy flow and autophagy-related proteins. MethodsSMMC7721 cells were treated with water extract of Jianpi Xiaoji prescription. CCK-8 assay was used to measure cell proliferation; flow cytometry was used to measure cell apoptosis rate; Western blot was used to measure the expression of the autophagy-related proteins Beclin1, LC3, and P62; the formation of autophagosomes and the fusion of autophagosomes and lysosomes were monitored. The independent samples t-test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsThe CCK-8 assay showed that cell proliferation was significantly inhibited at 24, 48, 72, and 96 hours after the drug was added, and there was a significant difference between the drug treatment group and the control group (t=28.458, 81.093, 85.328, and 100.158, all P＜0.001). Flow cytometry showed that compared with the control group, the drug treatment group had a significant increase in the apoptosis of liver cancer SMMC7721 cells (t=-42.629, P＜0.001). Western Blot showed that compared with the control group, the drug treatment group had significantly downregulated expression of Beclin1 and significantly upregulated expression of the autophagy markers LC3-II and P62. The flow of autophagosomes to autolysosomes was blocked in the drug treatment group, and there was a significant difference between the drug treatment group and the control group (F=31.155, P＜0.001). ConclusionThe water extract of Jianpi Xiaoji prescription can inhibit the proliferation of human liver cancer SMMC-7721 cells and promote apoptosis; meanwhile, it can upregulate the expression of Beclin1, downregulate the expression of LC3 and P62, and block the formation of autophagy flux.