The aim of the present study was to investigate the effect of bavachin treatment on A375 cells and the regulation of melanin synthesis. The cultured A375 cells in vitro were treated with bavachin; and the effect of bavachin on cell activity, tyrosinase (TYR) activity and melanin synthesis were respectively tested by the MTT assay, L-dopa oxidation assay and the NaOH lysis assay. The expression levels of TYR and c-Jun N-terminal kinases (JNK) proteins were tested by western blot analysis. The expression levels of TYR, tyrosinase-related protein-1 (TRP-1), TRP-2, extracellular signal-regulated kinase 1 (ERK1), ERK2 and JNK2 mRNA were tested by the reverse transcription-polymerase chain reaction assay. Simultaneously, the effect of estrogen receptor inhibitor (ICI182780) and ERK pathway inhibitor (U0126) was also tested on A375 cells following bavachin. The safe dose of bavachin significantly inhibited melanin synthesis and TYR activity. Bavachin (10 µmol/l) inhibited the expression of TYR and JNK proteins, and the expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2 mRNA in A375 cells. ICI182780 and U0126 could significantly reverse the bavachin treatment on the protein expression levels and the mRNA expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2. In conclusion, bavachin inhibited the synthesis of melanin on A375 cells by inhibiting the protein and mRNA expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2.