丹参酮ⅡA与阿霉素联合应用对人骨肉瘤MG63细胞增殖体外抑制作用

Inhibition Effect of TanshinoneⅡA Combined with Adriamycin on the Proliferation of Human MG63 Osteosarcoma in vitro

目的:研究丹参酮ⅡA(TanⅡA)与阿霉素(ADM)联合应用对人骨肉瘤MG63细胞增殖的抑制作用及机理。方法:不同浓度的TanⅡA与ADM联合处理MG63细胞后,MTT比色分析法检测细胞的增殖抑制率,流式细胞仪测定细胞的凋亡率,蛋白质印记法(Western blot)检测细胞Bcl-2,Bax和Caspase-3的蛋白表达水平改变。结果:处理48hADM+低浓度TanⅡA组与ADM+高浓度TanⅡA组的抑制率分别为62.35%±1.58%、76.38%±2.01%,明显高于ADM组(32.67%±1.65%)、低浓度TanⅡA组(18.55%±0.77%)和高浓度TanⅡA组(44.25%±1.01%);流式结果显示空白对照组、ADM组、低浓度TanⅡA组、高浓度TanⅡA组、ADM+低浓度TanⅡA组、ADM+高浓度TanⅡA组48h细胞凋亡率分别为0.03%,10.24%,4.47%,13.06%,13.23%,21.36%;联合用药组Bcl-2蛋白表达明显较单一用药组下调,而Bax和Caspase-3的蛋白表达较单一用药组明显上调。结论:TanⅡA与ADM联合应用对人骨肉瘤MG63细胞有显著的抑制作用,其作用可能与干预Bcl-2,Bax和Caspase-3等癌基因表达从而诱导细胞凋亡有关。

Objective：To investigate the effects and mechanism of combination of Tanshinone（Tan）ⅡA and adriamycin on the proliferation inhibition in human MG-63 Osteosarcoma.Methods：The MG-63 cells were treated with different concentrations of Tanshinone ⅡA and adriamycin.Cell growth was analyzed with MTT assay.Cell apoptosis was tested by flow cytometry.Westernblot was used to determine the protein expression of Bcl-2,Bax and Caspase-3of the MG-63 Osteosarcoma cells.Results：The 48 hours inhibition rate of adriamycin with low concentration TanⅡ A group and adriamycin with high concentration TanⅡ A group were 62.35% ±1.58% and76.38%±2.01%,which were higher than those in the TanⅡ A group and two adriamycin groups as 32.67%±1.65%,18.55%±0.77%and 44.25%±1.01%,respectively.The inhibition rate of each group was 0.03%,10.24%,4.47%,13.06%,13.23%,and 21.36%,respectively,by flow cytometry.Compared with the single drug group,down-regulated expression of Bcl-2and up-reg-ulated expression of Bax and Caspase-3were detected in the combination group.Conclusion：The combination of TanⅡ A and adriamycin has significant inhibitory effect on the human MG-63 Osteosarcoma cells.The role may be related to the intervention of Bcl-2,Bax and Caspase-3and other cancer relevant genes,which can bring induction of apoptosis.