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目的:探讨去甲斑蝥素抑制人骨肉瘤U2OS细胞生长的作用及其分子机制,为开发抗骨肉瘤新药提供实验基础。方法:以不同浓度的去甲斑蝥素处理骨肉瘤U2OS细胞,应用MTT法检测细胞生长抑制率,用Bliss法计算IC50值,通过流式细胞术和Hoechst33258荧光染色检测细胞凋亡,Caspase-3、8、9活性检测试剂盒检测其活性。结果:5、10、20、40、80μg/mL的去甲斑蝥素明显抑制骨肉瘤U2OS细胞生长并呈剂量依赖关系,48h的IC50值为26.15μg/mL;20μg/mL去甲斑蝥素作用12、24、48h诱导U2OS细胞的凋亡率分别为:7.8%、26.3%和43.9%,并出现明显的凋亡特征性形态变化;去甲斑蝥素处理U2OS细胞48h后Caspase-3、8、9的活性明显增强。结论:去甲斑蝥素可通过激活Caspase级联活化而抑制骨肉瘤U2OS细胞生长并诱导凋亡。
Objective: To investigate the effect of norcantharidin on the growth of human osteosarcoma U2OS cells and its molecular mechanism, providing experimental basis for the development of new anti-osteosarcoma drugs. Methods: The osteosarcoma U2OS cells were treated with different concentrations of norcantharidin. The cell growth inhibition rate was determined by MTT assay. The IC50 values were calculated by Bliss method. The apoptosis, Caspase-3 and CD44 expression were detected by flow cytometry and Hoechst33258 staining. 8,9 activity test kit to test its activity. Results: Norcantharidin 5,10,20,40,80μg / mL significantly inhibited the growth of osteosarcoma U2OS cells in a dose-dependent manner, with an IC50 of 26.15μg / mL at 48h and a dose of 20μg / mL norcantharidin , The apoptotic rates of U2OS cells induced by 24,48h were 7.8%, 26.3% and 43.9%, respectively. The apoptotic morphological changes of U2OS cells were observed. After treated with norcantharidin for 48h, the expressions of Caspase-3, The activity was significantly enhanced. CONCLUSION: Norcantharidin can inhibit the growth of osteosarcoma U2OS cells and induce apoptosis by activation of Caspase cascade.