Ethyl acetate subfractions from ethanol extracts of fermented oats (Avena sativa L.) exert anti-cancer properties in vitro and in vivo through G2/M and S Phase arrest and apoptosis

Background: Cancer is a major public problem and poses a long-term impact on patients' life, work, and study. Oats are widely recognized as healthy food and fermented oats were rich in the higher contents of polyphenols. However, the role of fermented oats in cancer remains elusive. Methods: The effect of ethyl acetate subfractions (EASs) from ethanol extracts of oats fermented by Rhizopus oryzae 3.2751 on cancer cells was verified by series experiments in vitro and in vivo. The cell viability, colony formation, cell cycle, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and western blot were determined in vitro. The toxicity of EASs and xenograft mouse model were performed in vivo. Results: MTT assay indicated that EASs interference suppressed the proliferation of four human cancer cells in a dose-dependent manner without a significant impact on two normal cells. EASs (0.2, 0.4, and 0.8 μg/mL) resulted in the G2/M and S phase arrest, apoptosis, depolarization of MMP, and ROS generation in HepG2 cells by flow cytometry. p53, JNK, caspase-9, and caspase-3 were activated and the expression of Bax was promoted, while the expression of Bcl-2 was reduced in HepG2 cells exposed to EASs via western blot. Furthermore, the in vivo study using a xenograft mouse model demonstrated that EASs attenuated the tumor growth with low systemic toxicity. Conclusions: EASs exhibited anti-cancer activities in vitro and in vivo via cell cycle arrest and apoptosis. This finding suggests that polyphenol-enriched composition from fermented oats might become a promising candidate for impeding the development and progression of liver cancer.