当归挥发油通过抑制PI3K/Akt/mTOR信号转导通路影响人结肠癌LOVO细胞自噬研究

目的 研究当归挥发油(volatile oil of Angelicae Sinensis Radix,VOAS)对人结直肠癌LOVO细胞自噬的影响及其分子机制。方法 用不同浓度的VOAS (6.25,12.5,25,50和100 μg·mL^-1)作用于人结肠癌LOVO细胞48 h,CCK8比色法检测VOAS对LOVO细胞增殖的抑制作用。将VOAS 50 μg·mL^-1作用于LOVO细胞,AO荧光染色和透射电镜观察细胞自噬情况;Western blotting观察VOAS对自噬相关蛋白LC3B、Beclin-1、Atg5及PI3K/Akt/mTOR信号转导通路的影响。结果 6.25~100 μg·mL^-1的VOAS能够抑制LOVO细胞的增殖,具有浓度依赖性;50 μg·mL^-1的VOAS能够促进LOVO细胞的自噬,上调自噬相关蛋白LC3B-Ⅱ、Beclin-1及Atg5的表达,同时下调PI3K、p-Akt、p-mTOR蛋白的表达。结论 VOAS能够促进人结肠癌LOVO细胞自噬,其机制可能与抑制PI3K/Akt/mTOR信号转导通路有关。

Study on Volatile Oil of Angelicae Sinensis Radix Affects Autophagy in Human Colorectal Cancer LOVO Cells by Inhibiting PI3K/Akt/mTOR Signal Transduction Pathway

OBJECTIVE To study the effect and mechanism of volatile oil of Angelicae Sinensis Radix(VOAS) on autophagy in human colorectal cancer LOVO cells. METHODS Human colorectal cancer LOVO cells were treated with various concentrations of VOAS(6.25, 12.5, 25, 50 and 100 μg·mL^-1) for 48 h, CCK8 assay was used to detect the effect of VOAS on the proliferation inhibition effect of cells. When treated with VOAS(50 μg·mL^-1), the effect of VOAS on autophagy was observed by acridine orange staining and electron microscopy, Western blotting was used to detect the expression of autophagy hallmark protein(LC3B, Beclin-1, Atg5) and PI3K/Akt/mTOR signal pathway. RESULTS The 6.25-100 μg·mL^-1 VOAS could significantly inhibit the proliferation of LOVO cells in dose-dependent manner, and VOAS(50 μg·mL^-1) could promote autophagy. Furthermore, VOAS(50 μg·mL^-1) could up-regulate the expression of LC3B-II, Beclin-1 and Atg5 protein that related to autophagy, and down-regulate the expression of PI3K, p-Akt and p-mTOR protein. CONCLUSION VOAS may induce autophagy in human colorectal cancer LOVO cells through inhibiting PI3K/Akt/mTOR signal transduction pathway.