人骨髓间质干细胞诱导成骨分化过程中差异表达蛋白的鉴定和分析  

Identification and analysis of differential expressed proteins in osteoblast differentiation from human bone marrow mesenchymal stem cells

作　　者：张爱霞[1] 余伟华[2] 王涛[1] 马保锋[2] 

机构地区：[1]嘉应学院生命科学学院生物工程系,广东梅州514015 [2]中山大学中山医学院干细胞与组织工程中心,广东广州510080

出　　处：《中国现代医学杂志》2015年第25期16-20,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(No:81000681;No:81000177)

摘　　要：目的探讨人骨髓间质干细胞(h MSCs)诱导成骨分化过程中的差异表达的蛋白点,为阐明h MSCs成骨分化的分子机制提供依据。方法收集h MSCs和成骨诱导7 d的细胞全蛋白,应用双向凝胶电泳和图像分析软件进行蛋白质点的识别和检测,应用比较蛋白质组学技术找出凝胶上的差异蛋白点,应用基质辅助激光解析离子化飞行时间质谱(MALDI-TOF-MS)和生物信息学方法对差异蛋白点进行蛋白质的鉴定和分析。结果通过MALDI-TOF-MS和生物信息学方法鉴定出了52种差异蛋白质,其中13种蛋白质在成骨诱导7 d后表达明显上调,39种蛋白质在成骨诱导7 d后表达明显下调;利用Gene Ontology对这些蛋白质按生物学过程和分子功能进行了分析。结论鉴定了52种可能在h MSCs诱导成骨分化过程中发挥重要作用蛋白质,这为进一步揭示h MSCs成骨分化的分子机制提供了科学依据。【Objective】To research human bone marrow mesenchymal stem cells（h MSCs） differentiation along osteoblasts to find the important functional proteins and elucidate the molecular mechanism in the process of differentiation into osteoblasts.【Methods】The total protein extracts were obtained with cell lysis buffer from undifferentiated h MSCs and osteogenic induced h MSCs on day 7. Using proteomic approaches based on two-dimensional gel electrophoresis（2-DE） and 2-DE gel analysis software, differently expressed protein spots were selected after being recognized and tested. The selected differently expressed protein spots were identified and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry（MALDI-TOF-MS） and bioinformatics. 【Results】Through MALDI-TOF-MS analysis and database searching, 52 proteins were identified including 13 up-regulated and 39 down-regulated in osteogenic induced h MSC for 7 days relative to undifferentiated h MSCs. The identified proteins were grouped by Gene Ontology according to biological process and molecular function. 【Conclusion】The identified differential proteins that maybe play important roles in osteogenic differentiation of h MSCs provide a comprehensive reference to understand molecular mechanism of osteogenic differentiation.

关 键 词：骨髓间质干细胞 成骨分化 蛋白质组学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]