Sulforaphane (SFN) is an organosulfur compound extracted from cruciferous vegetables and has biological effects. The effect of SFN has been studied in different types of cancers, as this compound incites various cytotoxic mechanisms to stunt cancer proliferation. However, the role of SFN activity in melanoma is yet to be known. The current study has been devised to elucidate the effects induced by SFN treatment in the B16F10 melanoma cell line and zebrafish model. Cells were treated with SFN reduced cell proliferation and increased tyrosinase production. Moreover, microscopic and immunofluorescence analysis confirmed the elongated appearance of melanoma cells due to cytoskeletal reorganization induced by SFN. Western blotting showed that SFN regulates the protein expression of Microphthalmia-associated transcription factor (MITF), Protein kinase C beta 1 (PKCβ1), and tyrosinase. The relationship between melanin biosynthesis and changes in the actin cytoskeleton encouraged by SFN on melanoma was determined by treating it with Cytochalasin D (CD) and Jasplakinolide (JAS). Co-treatment of SFN with CD increased more tyrosinase expression than SFN alone whereas with JAS, slightly reduced the expression. Immature zebrafish were pretreated with phenylthiourea (PTU) and then exposed to different SFN concentrations yielded the same results by upregulating the melanin levels despite the presence of melanin inhibitor (PTU). These study results show that SFN induces the biosynthesis of melanin in the B16F10 melanoma cell line, which occurs through changes in actin.