The aim of this work was to investigate the anticancer cytotoxic effects of natural compound subamolide E on the human skin cancer melanoma A375.S2 cells. Subamolide E was isolated from Cinnamomum subavenium and demonstrated cytotoxicities in the cell-growth assay at concentration ranges from 0 to 100 μM at 24 h. Propidium iodide staining and flow cytometry analyses were used to evaluate cell-cycle distribution and found that subamolide E caused DNA damage in the sub-G1 phase with a dose-dependent manner after 24 h of treatment. According to the western blot result, subamolide-E-treated cells with the increase of caspase-dependent apoptotic proteins induced related pathway mechanisms. Subamolide E also showed antimigratory activities of A375.S2 cells on the wound-healing assay. Finally, subamolide E demonstrated minor cytotoxicities to normal human skin cells (keratinocytes, melanocytes, and fibroblasts); therefore, it is a potential chemotherapeutic agent against skin melanoma.