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目的:探讨小檗胺体外对大鼠骨肉瘤UMR-106细胞增殖和凋亡的作用及机制。方法:用不同浓度的小檗胺(0,2,4,8,16,32mg.L-1)分别处理UMR-106细胞,作用时间分别为24,48,72h,采用四甲基偶氮唑盐(MTT)比色法检测小檗胺对UMR-106细胞增殖的抑制作用。用不同浓度的小檗胺(0,4,8,16mg.L-1)分别处理UMR-106细胞24h后,采用流式细胞术(FCM)检测细胞凋亡率(Annexin/PI染色),采用酶联免疫吸附试验(ELISA)检测细胞内Bcl-2、Bax水平,采用分光光度法检测caspase-3的活性。结果:小檗胺对UMR-106细胞的作用呈时间-剂量依赖性,孵育72h、32mg.L-1的小檗胺对UMR-106细胞的作用最强,抑制率接近90%;24,48,72h的IC50分别为24.69、8.03和3.54mg.L-1。空白对照组和小檗胺处理组细胞凋亡率分别为(1.64±0.29)%,(3.58±0.31)%,(6.3±0.5)%和(11.3±1.1)%。同时,小檗胺可以诱导细胞坏死;此外,小檗胺剂量依赖性地降低Bcl-2,增高Bax,降低Bcl-2/Bax比值,并增强caspase-3的活性。结论:小檗胺体外能抑制UMR-106细胞增殖并诱导细胞凋亡或坏死,其诱导凋亡的机制可能与降低Bcl-2/Bax比值,增强caspase-3的活性有关。
Objective: To investigate the effects and mechanisms of berbamine on proliferation and apoptosis of rat osteosarcoma UMR-106 cells in vitro. Methods: UMR-106 cells were treated with different concentrations of berbamine (0, 2, 4, 8, 16, 32 mg.L-1) for 24, 48 and 72 h, respectively. Salt (MTT) colorimetric assay berbamine inhibit the proliferation of UMR-106 cells. After treated with various concentrations of berbamine (0, 4, 8, 16 mg.L-1) for 24 h, the apoptotic rates of the cells were detected by flow cytometry (FCM) Enzyme-linked immunosorbent assay (ELISA) was used to detect intracellular levels of Bcl-2 and Bax, and the activity of caspase-3 was detected by spectrophotometry. Results: The effect of berbamine on UMR-106 cells in a time-and-dose-dependent manner, incubation 72, 32mg.L-1 berbamine strongest effect on UMR-106 cells, the inhibition rate of nearly 90%; 24,48 , 72h’s IC50 were 24.69,8.03 and 3.54mg.L-1. The apoptotic rates of the blank control group and berbamine group were (1.64 ± 0.29)%, (3.58 ± 0.31)%, (6.3 ± 0.5)% and (11.3 ± 1.1)%, respectively. At the same time, berbamine induced cell necrosis; in addition, berbamine dose-dependently decreased Bcl-2, increased Bax, decreased Bcl-2 / Bax ratio, and enhanced caspase-3 activity. Conclusion: Berbamine can inhibit the proliferation of UMR-106 cells and induce apoptosis or necrosis in vitro. The mechanism of apoptosis induced by berbamine may be related to the decrease of Bcl-2 / Bax ratio and the increase of caspase-3 activity.