Atractylon induces autophagy-dependent apoptosis in hepatocellular carcinoma cells via inhibition of the PI3K/AKT/mTOR pathway.

Hepatoma is one of leading cancer-related deaths. Regulation of apoptosis and autophagy represents an effective strategy for treatment of hepatoma. Atractylon is a primary component isolated from Atractylodes macrocephala. The effects of atractylon on hepatoma cell fate and the underlying mechanisms were investigated. Here, HepG2 cells were cultured and treated with atractylon. Cell proliferation, apoptosis, autophagy, and related signaling pathway were examined using various cellular and molecular approaches. Atractylon treatment for 24 h reduced viability and inhibited proliferation in HepG2 cells. Atractylon also stimulated mitochondrial apoptosis evidenced by Tunel positive staining and loss of mitochondrial membrane potential (MMP), which was accompanied by downregulation of Bcl-2 and upregulation of Bax and by caspase-9 and -3 activation in HepG2 cells. The phosphoinositide 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway was inhibited by atractylon treatment for 6 h in HepG2 cells, leading to the induction of mitochondrial apoptosis. Atractylon treatment for 12 h activated autophagic flux, because atractylon-induced autophagy was abolished by 3-methyladenine but was enhanced by chloroquine or bafilomycin A1. Furthermore, loss of MMP and activation of caspases upon atractylon treatment were abrogated by 3-methyladenine or autophagy-related gene 3 (ATG3) siRNA in HepG2 cells, suggesting that autophagy activation was required for induction of apoptosis. Altogether, atractylon disrupted the PI3K/AKT/mTOR signaling leading to autophagy-dependent apoptosis, which could be a promising candidate for anti-hepatoma therapy.