Differentiation Induced by Ginsenoside Rg1 Combined Formula Involved the Alteration of Prohibitin Localization and Expression in Human Osteosarcoma MG-63 Cells
选择性抽提经人参皂苷Rg1组合(RCT)诱导处理前后的人成骨肉瘤MG-63细胞核基质,对prohibitin在核基质中的存在、分布及其与相关基因产物在RCT处理前后MG-63细胞中的共定位关系进行观察研究.蛋白质组学分析结果显示,prohibitin存在于人成骨肉瘤MG-63细胞核基质蛋白组分中,并在RCT处理后细胞核基质中表达下调;蛋白质印迹杂交确证了prohibitin在MG-63细胞核基质中的存在及其在RCT处理后下调变化;免疫荧光显微镜观察进一步证实prohibitin定位在核基质上,经RCT处理后出现分布位置与表达水平变化;激光共聚焦显微镜观察可见prohibitin与c-Fos、c-Myc、p53和Rb基因产物均存在共定位关系,并在RCT处理后共定位分布区域出现变化.本研究证实了prohibitin是一种新发现的核基质蛋白,其在核基质上的定位与表达在RCT诱导分化前后发生显著变化,并与相关癌基因、抑癌基因产物存在共定位关系.实验表明RCT处理引起的prohibitin的变化与人成骨肉瘤MG-63细胞的诱导分化与调控具有密切关系,为深入揭示RCT等中药有效成分诱导肿瘤细胞分化的机理提供了重要科学依据和深入探索的新方向.Prohibitin is a nuclear matrix protein and associates with various oncogene products.A combined formula with ginsenoside Rg1,cinnamic acid and tanshinoneⅡA(RCT)was used to treat human osteosarcoma MG-63 cells in this study.We found that the prohibitin expression in the nuclear matrix was of decreased following RCT treatment by Western blot.The colocalization of prohibitin with the products of oncogenes or tumor suppressor genes,including c-Fos,c-myc,p53 and Rb was evaluated by immunofluorescence staining and laser scanning confocal microscopy.We observed significant alternations of prohibiitin distribution and the degree of colocalization with the associated gene products after the cells exposed to RCT.The results suggested that prohibitin might be involved in the RCT-induced differentiation in MG-63 cells.Our findings may provide information to expand the applications of Traditional Chinese Medicine formulas in cancer-related studies.国家自然科学基金资助项目(No.30470877)~
