去甲斑蝥素抑制人骨肉瘤U2OS细胞生长及其机制研究

The Growth Inhibition Role of Norcantharidin on human osteosarcoma U2OS cells

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 study the growth inhibition effect of norcantharidin on human osteosarcoma U2OS cells and its molecular mechanisms.Meanwhile,it provide experimental basis for developing new anti-osteosarcoma drugs.Methods： U2OS cells were treated with different concentrations of norcantharidin and then detect growth inhibition rate with MTT assay.IC50value was calculated with Bliss method and cell apoptosis was detected with flow cytometry and Hoechst33258 fluorescence staining.The activities of caspase-3,8,9were assayed by caspase colorimetric assay kit.Results：At concentrations of 5、10、20、40、80μg/ml,norcantharidin can inhibit U2OS cells growth in a dose-dependent manner.The IC50value was 26.15μg/ml 48hlater.Treated with 20μg/ml norcantharidin for 12,24and 48h,the apoptosis rate of U2OS cells reached 7.8%,26.3%and 43.9%respectively,and the typical morphological changes occurred.The activities of caspase3,8,9in U2OS cells increased obviously 48hafter the treatment with norcantharidin.Conclusion：Norcantharidin induces growth inhibition and apoptosis in human osteosarcoma U2OS cells by caspase activating as a cascade