Objective To explore the effects of Neferine on the proliferation and metastasis of gastric cancer cells in vivo and in vitro. Methods MTT assay, 5-ethyl-2′-deoxyuridine (EdU) incorporation assay, cell scratch assay and Transwell assay were used to detect the proliferation, migration, and invasion of gastric cancer cells, respectively. Western blot was used to detect the expression of cell proliferation and metastasis related proteins. In vivo, a xenograft tumor model was constructed by subcutaneous injection in nude mice, a lung metastasis model was constructed using tail vein injection. Immunohistochemistry was used to detect the protein levels of proliferation related markers and metastasis related markers.Results Compared with Neferine 0 μM group, Neferine 5 μM and 10 μM treatment significantly inhibited the proliferation, migration, and invasion of gastric cancer cells in vitro. Compared with Neferine 0 μM group, Neferine 5 μM and 10 μM treatment significantly reduced Ki67 and MMP2 protein expression, while increased TIMP2 protein expression. In vivo, compared with the control group, Neferine significantly inhibited the tumor growth and lung metastasis. Conclusion Neferine has strong anti-tumor activity and is a promising candidate drug for the treatment of gastric cancer