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

发布时间：2018-11-13 18:13

【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。
[Abstract]:Objective to compare the proteome differences of human bone marrow mesenchymal stem cells (BMSCs) during osteogenic differentiation induced by (h MSCs), and to search for the related proteins of h MSCs osteogenic differentiation. Methods MSCs was cultured in vitro and osteogenic differentiation was induced. The whole proteins of the cells with h MSCs and osteogenic differentiation for 3 days were collected. The protein spots were identified and detected by two dimensional gel electrophoresis (2-DE) separation and image analysis software. By comparing proteomics techniques, we found the protein spots on the gel that were 2.0 times different. The differential protein spots were identified and analyzed by matrix assisted laser ionization time of flight mass spectrometry (MALDI-TOF-MS) and bioinformatics. Results h MSCs successfully induced osteogenic differentiation in vitro. 38 differentially expressed proteins were identified by 2-DE isolation, MALDI-TOF-MS analysis and bioinformatics, of which 22 proteins were up-regulated 3 days after osteogenesis induction. The expression of 16 proteins was down-regulated 3 days after osteogenesis induction. According to the molecular function and biological pathway, the proteins identified by Gene Ontology showed that the proteins involved in metabolism, development, catalytic reaction and enzyme regulation activities accounted for 2932% and 444.16%, respectively. Conclusion Proteomics shows the difference of protein expression in the process of h MSCs induced osteogenesis, which provides a new idea for further elucidating the molecular mechanism of h MSCs osteogenic differentiation.
【作者单位】：嘉应学院生命科学学院;中山大学中山医学院干细胞与组织工程中心;
【基金】：国家自然科学基金(编号:81000681、81000177)
【分类号】：R329.2

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