Abstract: Objective To investigate the effect of Pinocembrin (Pin) on apoptosis and migration of mouse melanoma B16 cells and its relationship with DAAM2/Wnt/β-catenin, signaling axis. Methods CCK8 method was used to screen the Chinese medicine monomers which could inhibit the viability of B16 cells in vitro. Flow cytometry and wound healing assay were used to detect the apoptosis and migration of melanoma B16 cells. RNA-seq sequencing combined with GEPIA database was used to analyze the possible core genes of Pin. The expression levels of DAAM2 and Wnt/β-catenin signaling pathway-related proteins Axin2, p-GSK3β, and p-β-catenin were detected by real-time fluorescence quantitative PCR and Western blot. Reversion experiments were performed by transfection of a DAAM2 overexpression plasmid. A mouse melanoma model was established in vivo, and the mice were treated with low and high doses (10 mg/kg, 30 mg/kg) of Pin by gavage. The growth of melanoma in mice was observed. Western blot was used to detect the effect of Pin on the expression of DAAM2, Axin2, β-catenin and p-GSK3β in melanoma tissues to further clarify the effect of Pin on the progression of melanoma in mice. Results Pin significantly promoted the apoptosis of B16 cells (p < 0.001) and inhibited the migration of B16 cells (p < 0.01, p < 0.001). RNA-seq and GEPIA analysis suggested that DAAM2 was a key target. The results of qPCR and Western blot showed that Pin could significantly reduce the mRNA and protein expression of DAAM2 (p < 0.05, p < 0.01, p < 0.001). The expression levels of Wnt/β-catenin signaling pathway-related proteins p-GSK3β, p-β-catenin and Axin2 were down-regulated (p < 0.001). In vivo experiments showed that high-dose Pin could significantly inhibit the growth and weight of melanoma in mice, and reduce the expression of DAAM2, Axin2, β-catenin and p-GSK3β(p < 0.05, p < 0.01) in tumor tissues. DAAM2 overexpression can reverse the effect of Pin on the DAAM2/Wnt/β-catenin signaling axis. Conclusion Pin can promote the apoptosis of melanoma cells, inhibit their migration, and inhibit the growth of melanoma in mice by down-regulating the DAAM2/Wnt/β-catenin signaling axis. Key words: Pinocembrin; Melanoma; Anti-tumor; DAAM2; Wnt/β-catenin