人骨髓间质干细胞诱导成骨分化蛋白质的鉴定及生物信息学分析

Protein identification and bioinformatics analysis of osteoblast differentiation from human bone marrow mesenchymal stem cells

摘要:

目的：通过蛋白质组学和生物信息学技术建立人骨髓间质干细胞（human bone marrow mesenchymal stem cells，hMSCs）诱导成骨分化蛋白表达谱，为进一步阐明 hMSCs 的成骨分化机制提供基础资料。方法：收集 hMSCs 诱导成骨分化的全蛋白，应用双向凝胶电泳（two-dimensional gel electrophoresis，2-DE）技术和图像分析软件进行蛋白质点的识别和检测，选择清晰表达的蛋白质点，应用基质辅助激光解析离子化飞行时间质谱（matrix-assisted laser desorption ionization-time of flight mass spectromety，MALDI-TOF-MS）和生物信息学方法进行蛋白质的鉴定和分析。结果：通过 MALDI-TOF-MS 和生物信息学鉴定了77种蛋白质；建立了hMSCs诱导成骨分化蛋白质表达谱；利用Gene Ontology对这些蛋白按分子功能和生物学途径进行了归类。结论：成功建立了hMSCs诱导成骨分化蛋白质表达谱；为进一步阐明 hMSCs 成骨分化的分子机制提供了基础资料，为进一步构建 hM－SCs 诱导成骨分化蛋白质表达数据库奠定了坚实的基础。

Abstract:

Objective：To establish protein expressing profile of human bone marrow mesenchymal stem cells（hMSCs） which were induced into osteoblasts through proteomic and bioinformatic techniques and to further elucidate the precise molecular mechanism of hMSCs osteoblast differentiation. Methods：The total protein extracts were obtained with cell lysis buffer from hMSCs which were induced into osteoblasts. Two-dimensional gel electrophoresis（2-DE） and 2-DE gel analysis software were used to recognize and test protein spots. Clearly expressed protein spots were selected and were identified and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectromety（MALDI-TOF-MS） and bioinformatics． Results：Protein expressing maps of hMSCs induced into osteoblasts were set up. Seventy-seven proteins were identified by MALDI-TOF-MS and bioinformatics，which were classified by Gene Ontology according to molecular function and biological process. Conclusion：Protein expressing maps of hMSCs induced into osteoblasts are built，which will provide basal information to illuminate the molecular mechanism of hMSCs differentiation into osteoblasts and to build protein expression database of osteoblast differentiation from hMSCs.