〔Abstract〕 Objective To investigate the effect of aloperine (Alo) on the proliferation, migration and invasion of gastric cancer cells. Methods Human gastric cancer cell lines HGC-27 and AGS were treated with 0, 100, and 200 μmol/L Alo. CCK-8 was used to detect cell viability. Colony formation assay was used to detect cell proliferation. Flow cytometry was used to detect apoptosis. Scratch and Transwell migration assays were used to detect cell migration. Transwell invasion assay was used to detect cell invasion. Western blot was used to detect the expression of proteins related to proliferation, apoptosis, migration, invasion and Hippo pathway. Results Compared with the control group, the cell viability, number of colony formation, cell migration and invasion were significantly reduced after 100 and 200 μmol/L Alo treatment, and the apoptosis rate significantly increased ( P < 0.05 ). Additionally, the expressions of proliferating cell nuclear antigen (PCNA), B-cell lymphoma/leukemia-2 protein (Bcl-2), neural cadherin (N-cadherin), Vimentin, and transcriptional co-activator with PDZ-binding motif (TAZ) were significantly reduced, whereas the expressions of Bcl-2-associated X protein (Bax), phosphorylated yes-associated protein (p-YAP), and large tumor suppressor 1/2 (LATS1/2) significantly increased ( P < 0.05 ). Conclusion Alo inhibits the proliferation, migration and invasion of gastric cancer cells HGC-27 and AGS by regulating the Hippo signaling pathway.