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目的:探讨植物钩吻中生物碱单体对肝癌细胞HepG2增殖的抑制作用,并探讨其相关机制。方法:MTT法观察3种生物碱单体对HepG2细胞的体外杀伤作用,选取活性最好的单体进一步行时效、量效关系的测定和细胞形态学观察;流式细胞术检测细胞周期和凋亡率的变化;Capspase-3、Capspase-8、Capspase-9活性检测用分光光度法进行。结果:3种单体都显示出对HepG2细胞的增殖抑制活性,其中钩吻素己效果最好,呈时间及剂量依赖性抑制HepG2细胞增殖;经钩吻素己处理48 h后的HepG2细胞变小变圆,正常贴壁细胞随着药物浓度的增加而减少;各浓度组钩吻素己可阻滞细胞于S期,并诱导细胞凋亡,且Capspase-3、Capspase-8、Capspase-9活性随着药物浓度的增高而增高。结论:3种钩吻生物碱单体中钩吻素己能显著抑制HepG2细胞体外增殖且细胞毒性较小,并通过影响细胞周期分布和激活Capspase-8、Capspase-9进而活化Capspase-3发挥抗肿瘤作用。
Objective: To investigate the inhibitory effect of alkaloid monomer in plant kiss on the proliferation of HepG2 hepatocellular carcinoma cells and to explore its mechanism. Methods: The killing effects of three alkaloids on HepG2 cells were observed by MTT method. The most active monomer was selected for further study on the effect of apoptosis, cell cycle and apoptosis by flow cytometry Changes in mortality; Capspase-3, Capspase-8, Capspase-9 activity was measured spectrophotometrically. Results: HepG2 cells showed inhibitory activity on all three kinds of cells. The best one was in the HepG2 cells, and the proliferation of HepG2 cells was inhibited in a dose and time dependent manner. Small round, normal adherent cells decreased with the increase of drug concentration; each concentration of kyoto-kinin could block cells in S phase and induce apoptosis, and Capspase-3, Capspase-8, Capspase-9 Activity increased with increasing drug concentration. CONCLUSION: Agrocyberin can significantly inhibit the proliferation and cytotoxicity of HepG2 cells in vitro, and can activate Capspase-3 by affecting the cell cycle distribution and activation of Capspase-8 and Capspase-9 Tumor action.