| dc.description.abstract | Antrodia cinnamomea, an endemic fungus of Taiwan, is well known as a herbal medicine to remedy various illnesses, including liver cancer, a major cause of death in Asia. However, the early mechanism of its anti-cancer effect was unclear. In this study, we explored the anti-cancer mechanism induced by Antrodia cinnamomea fruiting body ethanol crude extract (AcFBE) on human hepatocellular carcinoma cell (SK-HEP-1) through in vitro assay in conjunction with miRNA and mRNA transcriptome profiling using next generation sequencing (NGS). Results indicated that 500μg /ml of AcFBE can induce apoptotic cell death on SK-HEP-1 within 2 hours, as characterized by cell viability assay, cell morphology alteration, cell membrane extroversion, caspases activation, and DNA fragmentation. The sequence data revealed 354 known miRNAs from miRNA libraries. To our surprise, 85.4% of known miRNAs in SK-KEP-1 were decreased, while only 57.3% of known miRNAs were found decreased for normal mouse liver cells (BNL CL.2). The AcFBE-affected miRNAs expression profiles showed dramatic down-regulation in SK-HEP-1 cells, with 95.2% (141/148) and 84.1% (122/145) affected after 2 hr and 4 hr AcFBE treatment, respectively, suggesting that AcFBE caused a global inhibition of miRNA level. This result was supported by western blotting showing decreases of miRNA biogenesis proteins, Dicer and Drosha, together with an increase of XRN2 involved in miRNA degradation. Transcriptome analysis showed 2077 (58.3%) and 1491 (40.1%) up-regulated genes in 2-hr and 4-hr libraries, respectively, based on 2 fold cutoff. We then analyzed 365 genes most significantly regulated by AcFBE using GO term analysis on transcriptome data, indicating a strong association with apoptosis. Overall, our study suggests that AcFBE is able to globally reduce SK-HEP-1 miRNA expression and thereafter affect gene expression regulation, causing cancer cell death by apoptosis. This is the first report implicating miRNA in the anti-cancer effects of A. cinnamomea fruiting body.
| en_US |