Objective To explore the effects of advanced glycation end product (AGE) on proliferation and differentiation of mouse embryonic osteoblast line MC3T3-E1 cells and its mechanisms.Methods MC3T3-E1 cells were treated with different mass concentrations of AGE (100, 200, and 300 mg/L). Cell proliferation activity and apoptosis rate were detected by cell counting kit 8 and flow cytometry, respectively. Osteogenic ability was detected by alkaline phosphatase (ALP) staining, and the mRNA expression of osteogenesis-related genes (osteocalcin, ALP, and Runx2), Yes-associated protein (YAP), and β-catenin was detected by quantitative polymerase chain reaction. The protein expression of YAP and β-catenin was detected by Western blotting, and the nuclear contents of YAP and β-catenin and the expression of cytoskeleton protein filamentous actin (F-actin) were observed by immunofluorescence assay.Results The proliferation activity of MC3T3-E1 cells was significantly decreased and the apoptosis rates were significantly increased after treatment with 200 or 300 mg/L AGE (all P < 0.05), and 100 mg/L AGE had no significant effect on cell proliferation or apoptosis and was selected for the experiment. Under osteogenic induction culture condition, compared with the control group, the ALP staining was shallow in MC3T3-E1 cells after treatment with 100 mg/L AGE, and the mRNA expression levels of osteocalcin, ALP and Runx2 were significantly lower (all P < 0.05). Under conventional culture condition, compared with the control group, the morphology and distribution of F-actin were significantly changed in MC3T3-E1 cells after treatment with 100 mg/L AGE; there were no significant changes in the mRNA or protein expression of YAP, but the nuclear contents were decreased; and the mRNA and protein expression levels of β-catenin were significantly decreased (both P < 0.05), but the nuclear contents had no significant change.Conclusion AGE can inhibit proliferation activity, induce apoptosis, and inhibit osteogenic differentiation ability of MC3T3-E1 cells. F-actin, YAP, and β-catenin participate in the regulation process.