人骨髓间质干细胞诱导成骨分化过程中差异表达蛋白的质谱鉴定及生物信息学分析  

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

作　　者：张爱霞[1,2] 余伟华[2] 王涛[1] 

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

出　　处：《安徽医科大学学报》2014年第12期1722-1726,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81000681;81000177)

摘　　要：目的比较人骨髓间质干细胞(h MSCs)诱导成骨分化过程中的蛋白质组差异,寻找h MSCs成骨分化相关蛋白质。方法体外培养h MSCs并诱导成骨分化,收集h MSCs和成骨诱导分化3 d的细胞的全蛋白,应用双向凝胶电泳(2-DE)分离和图像分析软件进行蛋白质点的识别和检测,通过比较蛋白质组学技术找出凝胶上差异2.0倍以上的蛋白质点,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 h MSCs在体外成功诱导成骨分化,通过2-DE分离、MALDI-TOF-MS分析和生物信息学方法鉴定出了38种差异蛋白,其中22种蛋白质在成骨诱导3 d后表达明显上调,16种蛋白质在成骨诱导3 d后表达明显下调。利用Gene Ontology对所鉴定的蛋白质按分子功能和生物学途径进行分析显示,参与体内代谢、发育过程、催化反应和酶调节活性的蛋白质分别占29%、32%、44%、16%。结论蛋白质组学较好地显示了h MSCs诱导成骨分化过程中的蛋白质表达差异,这为进一步阐明h MSCs成骨分化的分子机制提供了新的思路。Objective To compare the proteomics discrepancies between human bone marrow mesenchymal stem cells （hMSCs） and its differentiation along osteoblasts to find the important functional protein candidates and elucidate the molecular mechanism in the process of differentiation into osteoblasts.Methods hMSCs were cultured and induced into osteoblast in vitro.The total protein extracts were obtained with cell lysis buffer from undifferentiated hMSCs and osteogenic induced hMSCs on day 3.Using proteomic approaches based on two-dimensional gel electrophoresis （2-DE） and 2-DE gel analysis software,differently expressed protein spots that had an altered abundance ratio of greater than 2.0 fold were selected after being recognized and tested.The selected different protein spots were identified and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectromety （MALDI-TOF-MS） and bioinformatics.Results hMSCs were induced into osteoblast in vitro successfully.The total protein extracts were separated effectively with 2-DE.By MALDI-TOF-MS analysis and database searching,38 proteins were identified including 22 up-regulated,16 down-regulated in osteogenic induced hMSC for 3 days relative to undifferentiated hMSCs.The identified proteins that participated metabolism,development process,catalytic reaction and enzyme regulation respectively possessed 29％,32％,44％ and 16％ according to molecular function and biological process by Gene Ontology analysis.Conclusion Proteomic analysis can identify the proteins with variance of osteogenic differentiation of hMSCs,meantime,provide a comprehensive reference to understand molecular mechanism of osteogenic differentiation.

关 键 词：骨髓间质干细胞 蛋白质组学 成骨分化 质谱鉴定 生物信息学 

分 类 号：R331.22[医药卫生—人体生理学]