Atractylenolide II induces G1 cell-cycle arrest and apoptosis in B16 melanoma cells

Ethnopharmacological relevance: Atractylenolide II (AT-II) is a sesquiterpene compound isolated from the dried rhizome of Atractylodes macrocephala (Baizhu in Chinese), which is traditionally prescribed for melanoma treatment by Chinese medicine practitioners. Our previous study showed that AT-II can inhibit B16 cells proliferation. Here we investigate the mechanistic basis for the anti-proliferative activity of AT-II in B16 melanoma cells. Materials and methods: Cell viability was examined by MTT assay. Cell cycle distribution and apoptosis were determined by flow cytometry. Protein expression was determined by Western blotting. Results: AT-II treatment for 48 h dose-dependently inhibited cell proliferation with an IC(50) of 82.3 mu M and induced G1 phase cell cycle arrest. Moreover, treatment with 75 mu M AT-II induced apoptosis. These observations were associated with the decrease of the expression of Cdk2, phosphorylated-Akt, phosphorylated-ERK and Bcl-2, the increase of the expression of phosphorylated-p38, phosphorylated-p53, p21, p27, and activation of caspases-8, -9 and -3. In addition, a chemical inhibitor of p53, PFT alpha, significantly decreased AT-II-mediated growth inhibition and apoptosis. Conclusions: We demonstrated that the G1-arresting and apoptotic effects of AT-II in B16 cells involve p38 activation as well as ERK and Akt inactivation, and the cytotoxic/apoptotic effects of AT-II are potentially p53 dependent. These findings provided chemical and pharmacological basis for the traditional application of Baizhu in melanoma treatment. (C) 2011 Elsevier Ireland Ltd. All rights reserved.