关键词: 东北刺人参, Falcarindiol, 制备HPLC, 结肠癌, Wnt/&beta, -catenin信号通路

Abstract: This study aimed to develop a rapid and efficient preparative method for falcarindiol (FAD) and investigate the relationship of the anti-proliferative effect of FAD on colorectal cancer HCT-116 cells and regulating cell cycle arrest related genes.The separation and purification were performed on silica gel chromatography and preparative HPLC in order to get FAD with high purity,which was identified by spectral data.MTS method was employed to test the cytotoxity of FAD on colorectal cancer HCT-116 cells.Subsequently,the cell cycle arrest,the mRNA level,protein expression of β-catenin,cyclin D1 and c-myc of HCT-116 cells were analyzed by flow cytometry,RT-qPCR and Western blotting,respectively.As the results,a rapid and efficient HPLC method has been validated for the preparation and purification of FAD from O.elatus.FAD showed cytotoxity on HCT-116 cells with the IC₅₀ values of 8.1±1.4,4.6±0.5 and 3.2±0.4 μmoL/L at 24,48 and 72 h,respectively,which could arrest the cell cycle of HCT-116 cell line at G2/M phase and significantly down-regulate the mRNA level and protein expressions of β-catenin,cyclin D1 and c-myc.These results suggested that FAD possessed a strong cytotoxic activity against HCT-116 cells,and arrested its cell cycle at G2/M phase,while its mechanism referred to Wnt/β-catenin signaling pathway.

Key words: Oplopanax elatus, falcarindiol, preparative HPLC, colorectal cancer, Wnt/&beta, -catenin signaling pathway