Background: Phosphoinositide 3-kinase/Protein kinase B (PI3K/AKT) pathway activation correlates with the clinical progress of melanoma. Previous research demonstrated the inhibitory effect of parthenolide on PI3K/AKT pathway. It was testified whether parthenolide could be utilized to inhibit PI3K-AKT pathway activation and relevant growth in melanoma in this study. Methods: Flow cytometry based on Annexin V/propidine iodide (PI) staining was used to measure cell apoptosis. A xenograft mouse model was constructed, and parthenolide was intraperitoneally injected to examine the effects of parthenolide in vivo. Parthenolide showed cytotoxicity on A375 and A2058 cells as revealed by diminishing viability, colony formation, and migration. Results: Increased apoptosis was found in A375 and A2058 cells induced by parthenolide treatment. Up-regulated Bax, and down-regulated B-cell lymphoma 2 (Bcl-2) expression was observed after parthenolide administration. Parthenolide also reduced the levels of phosphorylation of AKT and PI3K. In addition, parthenolide inhibited the growth of melanoma in the xenograft model. Conclusions: In summary, parthenolide inhibits cell viability and induces apoptosis in melanoma through the inhibition of PI3K/AKT signaling pathway, which could be considered an adjuvant therapy.