云南美登木提取物抑制胃癌的药理作用研究

Study on pharmacological action of Yunnan Maytenus Hookeri Loes.Extract on inhibiting gastric cancer

目的:评价云南美登木提取物(MDM)在体内外抑制胃癌的作用效果,并对其作用机制进行探讨。方法:采用细胞计数试剂盒(CCK-8)、活死染色、克隆形成、流式细胞术及高内涵成像等方法研究MDM对胃癌细胞SGC7901及MGC-803增殖、凋亡、周期、线粒体膜电位及活性氧(ROS)生成的影响;通过转录组学及蛋白组学对MDM调控的基因及蛋白进行分析,确定调控通路后采用Western Blot法进行验证;构建裸鼠皮下肿瘤模型评价MDM体内抗胃癌活性。结果:CCK-8、活死染色及克隆形成实验显示,MDM可时间和剂量依赖性地显著抑制胃癌细胞的增殖;流式细胞术及高内涵成像结果显示,MDM可诱导细胞凋亡,阻滞细胞周期于G2/M期,同时增加胞内ROS水平,降低线粒体膜电位。转录组及蛋白组的结果显示,MDM主要影响周期凋亡相关信号通路,Western blot实验得到相同结果。动物实验显示,MDM在体内可高效抑制胃癌生长。结论:MDM在体内外均具有良好的抗胃癌作用,且其可激活p53信号通路,产生氧化应激损伤,导致线粒体膜电位去极化,诱导胃癌细胞凋亡。

Objective:To evaluate the inhibitory effect of Yunnan Maytenus Hookeri Loes.extract(MDM)on gastric cancer in vitro and in vivo,and explore the mechanism of action.Methods:The effects of MDM on the proliferation,apoptosis,cell cycle,mitochondrial membrane potential and reactive oxygen species(ROS)generation of gastric cancer cells SCC7901 and MCC-803 were studied by CCK-8,Calcein AM/PI staining,colony formation assay,flow cytometry and high-content imaging.Subsequently,the genes and proteins regulated by MDM were analyzed by transcriptomics and proteomics,and the regulatory pathway was verified by Western Blot after determined.Finally,nude mice with subcutaneous tumor were modeled to evaluate the anti-gastric cancer activity of MDM in vivo.Results:CCK-8,Calcein AM/PI staining and colony formation assay showed that MDM significantly inhibited the proliferation of gastric cancer cells in a time-and dose-dependent manner.The results of flow cytometry and high-content imaging showed that MDM induced apoptosis,arrested the cell cycle in G,/M phase,increased intracellular ROSs level,and decreased mitochondrial membrane potential.The results of transcriptome and proteome showed that MDM primarily affected signaling pathways related to cell cycle and apoptosis,which was consistent with the results of Western Blot.Animal experiments showed that MDM effectively suppressed the growth of gastric cancer in vivo.Conclusion:MDM exhibits potent anti-gastric cancer effect both in vitro and in vivo.It can active p53 signaling pathway to produce oxidative stress damage,leading to depolarization of mitochondrial membrane potential,which induces apoptosis of gastric cancer cells.