目的：探索南蛇藤提取物齐墩果烷型五环三萜（28-hydroxy-3-oxoolean-12-en-2-oic acid）协同miR-451对人胃癌AGS细胞增殖、迁移的影响及其可能的分子机制。方法：用miR-451过表达慢病毒感染AGS细胞，并用盐酸多西环素（DOX）10或100 ng/ml 诱导 24 h，构建过表达 miR-451 的细胞 AGS/miR-451+。采用 10、20、40、80、160 μmol/L 的齐墩果烷型五环三萜处理AGS/miR-451+细胞，MTT法、划痕实验分别检测细胞增殖和迁移能力的变化，WB法检测细胞中mTOR通路及凋亡相关蛋白表达水平的变化。结果：成功构建过表达miR-451的AGS/miR-451+细胞。与未加药对照组相比，齐墩果烷型五环三萜处理后AGS/miR-451+细胞的增殖抑制率均呈时间和浓度依赖性升高（P<0.05或P<0.01），细胞迁移率均显著降低（P<0.05或P<0.01）。齐墩果烷型五环三萜处理组细胞中，mTOR 信号通路相关蛋白的表达量均有所降低（P<0.05或P<0.01）；凋亡相关蛋白中，Bcl2的表达量下降，BAX、caspase-3、caspase-1及细胞色素c的表达量升高（P<0.05或P<0.01）。结论：齐墩果烷型五环三萜能够协同miR-451抑制人胃癌AGS细胞的增殖与迁移，其机制可能与影响凋亡和mTOR信号通路相关蛋白的表达有关。

Objective: To explore the synergistic effect of 28-hydroxy-3-oxoolean-12-en-2-oic acid and miR-451 on the proliferation and migration of human gastric cancer AGS cells and its possible molecular mechanism. Methods: AGS cells were infected with miR-451 overexpression lentivirus and induced with DOX (10 or 100 ng/ml) for 24 h to construct AGS/miR-451+ cells overexpressing miR-451. AGS/miR-451+ cells were treated with 28-hydroxy-3-oxoolean-12-en-2-oic acid of 10, 20, 40, 80, 160 μmol/L. The changes in proliferation and migration ability of cells were detected by the MTT method and the scratch test, respectively; WB method was used to detect the changes in the expression levels of mTOR signaling pathway-related and apoptosis-related proteins in cells. Results:AGS/miR-451+ cells overexpressing miR-451 were successfully constructed. Compared with the untreated control group, the proliferation inhibition rate of AGS/miR-451+ cells in the 28-hydroxy-3-oxoolean-12-en-2-oic acid treatment group increased in a time[1]and concentration-dependent manner (P<0.05 or P<0.01), while the cell migration rate was significantly reduced (P<0.05 or P<0.01). In the cells treated with 28-hydroxy-3-oxoolean-12-en-2-oic acid, the expression of mTOR signaling pathway-related proteins was reduced (P<0.05 or P<0.01); among the apoptosis pathway-related proteins, the expression of Bcl2 was decreased, but the expression of BAX,caspase-3, caspase-1, and cytochrome c was increased (P<0.05 or 0.01). Conclusion: The 28-hydroxy-3-oxoolean-12-en-2-oic and miR-451 can synergistically inhibit the proliferation and migration of human gastric cancer AGS cells. The mechanism may be related to their regulation of apoptosis signaling pathway- and mTOR signaling pathway-related proteins.

江苏省自然科学基金委面上项目资助（No. BK20171290）；江苏省教育厅自然科学基金重大项目资助（No. 19KJA480003）