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小檗胺对大鼠骨肉瘤UMR-106细胞增殖的抑制作用

王巨存1,胡永成1,潘振华2,李正翔2,胡人杰3   

  1. 1. 天津市天津医院
    2. 天津医科大学总医院
    3. 天津市医药科学研究所
  • 收稿日期:2011-12-07 修回日期:2012-03-31 出版日期:2012-10-15 发布日期:2012-10-15
  • 通讯作者: 王巨存

Inhibitory Effect of Berbamine on Proliferation of Rat Osteosarcoma Cells UMR-106

  • Received:2011-12-07 Revised:2012-03-31 Published:2012-10-15 Online:2012-10-15
  • Contact: WANG Ju-Cun

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摘要： 目的：探讨小檗胺体外对大鼠骨肉瘤UMR-106细胞增殖的抑制作用及机制。方法：0、2、4、8、16、32 mg/L小檗胺作用UMR-106细胞24、48、72 h后，采用四甲基偶氮唑蓝（MTT）比色法检测小檗胺对UMR-106细胞增殖的抑制作用；0、4、8、16 mg/L小檗胺作用UMR-106细胞24 h后，Hoechst33258染色激光共聚焦扫描显微镜观察细胞的形态学变化；Annexin V/碘化丙啶（PI）荧光标记流式细胞术（FCM）检测细胞凋亡率；PI荧光标记FCM检测细胞周期变化。结果：小檗胺以剂量依赖方式显著抑制UMR-106细胞的增殖（P < 0.01），其作用24、48、72 h的半数抑制浓度（IC50）分别为24.69、8.03和3.54 mg/L。小檗胺处理组可见核固缩和凋亡小体。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）%、（6.27±0.47）%和（11.27±1.09）%，与空白对照组比较，差异有统计学意义（P <0.01）；同时，小檗胺可以诱导细胞坏死；此外，与空白对照组比较，G0/G1期细胞比例增高，而S期和G2/M期细胞比例降低（P < 0.01）。结论：小檗胺体外能够抑制骨肉瘤UMR-106细胞增殖，其机制与诱导细胞凋亡、坏死和G0/G1期阻滞有关。

关键词: 小檗胺, 骨肉瘤, 细胞增殖, 细胞凋亡, 细胞周期, 体外研究, 剂量效应关系, 药物

Abstract: Objective: To explore the inhibitory effect of berbamine on the proliferation of UMR-106 cells and its possible mechanism in vitro. Objective: To explore the inhibitory effect of berbamine on the proliferation of UMR-106 cells and its possible mechanism in vitro. Methods: UMR-106 cells were cultured with different concentrations of berbamine （0,2,4,8,16 and 32 mg/L） for 24, 48 and 72 h ,respectively,and the proliferation were colorimetrically assayed by MTT. UMR-106 cells were interfered with different concentrations of berbamine （0,4,8 and 16 mg/L）for 24h,the morphological changes of UMR-106 cells were observed by laser confoca1 scan microscope after Hoechst33258 staining, the apoptotic rates were detected by flow cytometry （FCM）using Annexin V/PI staining, the cell cycle were detected by FCM using PI staining. Results: Berbamine could obviously inhibit the proliferation of UMR-106 cells in a dose-dependent manner （P < 0.01）.The IC50 value at 24,48 and 72h were 24.69、8.03 and 3.54 mg/L, respectively. There were nuclear condensation and apoptotic body in berbamine treated group. Apoptotic rates of control group and berbamine treated group were （1.64±0.29）% ，（3.58±0.31） %，（6.27±0.47） % and （11.27±1.09） %，respectively.There was a significant difference in apoptotic rate compared with the control group （P < 0.01）. Meanwhile, berbamine can induce cell necrosis .In addition, berbamine significantly increased the percentages of cells in G0/G1 phase and decreased in S phase and G2/M phase compared with the control group （P <0.01）. Conclusion:berbamine can inhibit the proliferation of UMR-106 cells in vitro, inducing cell apoptosis , necrosis and G0/G1 phase arrest may be the mechanism .

Key words: berbamine , osteosarcoma , cell proliferation, apoptosis, cell cycle, in vitro , dose-response relationship, 药物