目的研究中药有效成分人参皂甙Rg1、肉桂酸CINN及丹参酮TanⅡA的组合(简称RCT),诱导人成骨肉瘤MG-63细胞分化过程中,细胞核基质构型与蛋白质组成的变化。方法应用流式细胞仪检测细胞周期变化,用四甲基偶氮唑蓝(MTT)法绘制细胞生长曲线,以选择性抽提整装光镜与电镜技术及蛋白质组学技术分析观察处理前后MG-63细胞核基质构型与蛋白质组成的变化。结果生长曲线显示,RCT处理后MG-63细胞的增殖受到明显抑制,增殖抑制率在第7天达到72.37%,细胞周期出现明显的G0/G1期阻滞。MG-63细胞核基质为典型的肿瘤细胞恶性表型,在RCT处理后发生了向正常细胞转变的恢复性变化。RCT处理后的细胞中多种核基质功能蛋白的表达发生了变化。结论RCT能显著抑制体外培养MG-63细胞的增殖活动,将细胞周期阻滞于G0/G1期,诱导核基质构型发生了显著的恢复性变化,并改变核基质蛋白的组成,对MG-63细胞具有显著的分化诱导作用。
Objective To explore the alteration of nuclear matrix proteins and the configurational changes of nuclear matrix (NM) system during differentiation of the osteosarcoma MG-63 ceils in vitro after being induced by the combination of ginsenoside Rgl, cinnamic acid and tanshinone Ⅱ A ( RCT). Methods Cell cycle was investigated by flow cytometry and the growth curve was drawn with MTT method. The cells treated with or without RCT were subjected to selective extraction method and prepared for whole mount electron microscopy observation. The nuclear matrix-intermediate samples were examined under light and transmission electron microscopes respectively. The nuclear matrix proteins were subjected to analysis of proteomic technique. Results The proliferstion of the MG-63 ceils treated with RCT were repressed markedly; the rate of repression was 72.37 % on the seventh day, and the cell cycle was blocked at G0/G1 phase. The typical malignant phenotype of nuclear matrix of MG-63 tumor cell were reversed to normal cells. The expression of many functional proteins of nuclear matrix were changed in the MG-63 cells after the treatment with RCT. Conclusion RCT can repress markedly the proliferation of MG-63 cells, block cell cycle, induce a restorational change to the nuclear matrix architecture similar to that of normal cells, and change the constitutes of nuclear matrix proteins. Thereby RCT have a notable effect on MG-63 cells differentiation.