【摘要】 选择性抽提经中药有效成分人参皂甙Rg1组合(简称RCT)诱导处理前后的人成骨肉瘤MG-63细胞核基质,对nucleophosmin(NPM)在核基质中的存在、分布及其与相关基因产物在MG-63细胞中的共定位关系进行了观察研究。双向凝胶电泳和质谱鉴定结果显示NPM存在于MG-63细胞核基质蛋白组分中,在RCT处理后细胞核基质蛋白中表达下调。蛋白质印迹杂交结果证实了NPM在RCT处理前后的MG-63细胞核基质蛋白中的存在及其表达下调变化。免疫荧光显微镜观察显示NPM定位于MG-63细胞核基质上,经RCT处理后出现分布位置与表达水平的变化。免疫荧光-激光扫描共聚焦显微镜的观察结果显示NPM在MG-63细胞中与c-fos、c-myc、RB、p53等基因产物具有共定位关系,并在RCT处理后细胞核内其共定位区域发生了变化。研究结果证实了NPM存在于核基质上,是一种核基质蛋白,其在RCT诱导人成骨肉瘤MG-63分化过程中的表达与分布变化及其与相关癌基因、抑癌基因的关系显然对MG-63细胞分化具有重要影响,这为深入揭示中药有效成分诱导肿瘤细胞分化的机制提供了重要科学依据和深入探索的新方向。更多还原

【Abstract】 To explore the existence, distribution of nucleophosmin (NPM) in the nuclear matrix of the osteosarcoma MG-63 cells and the co-localizational relationship between NPM and the products of some interrelated genes, the MG-63 cells treated with or without the combination of ginsenoside Rg1, cinnamic acid and tanshinone II A (RCT) were selectively extracted and subjected to proteomic methodologies and laser scanning confocal microscopy (LSCM). 2-D PAGE, MS and Western blotting methods confirmed the existence of NPM and its decreased expression in the nuclear matrix proteins. LSCM observation showed that there were altered co-localization after RCT treatment between NPM and the product of genes c-fos, c-myc, RB and p53, respectively. Our study confirmed the existence of NPM in the nuclear matrix and the important role its expression and distribution played during the differentiation of MG-63 cells induced by effective ingredients from Chinese materia medica.更多还原