骨髓间充质干细胞与视网膜色素上皮细胞的蛋白质组学比较

Proteomics Analysis between Bone Marrow Mesenchymal Stem Cells and Retinal Pigment Epithelium

目的采用蛋白质组学的方法比较骨髓间充质干细胞(BMSCs)和视网膜色素上皮细胞(RPE)中蛋白表达的差异。方法对BMSCs和RPE细胞采用2D-DIGE双向电泳技术获取差异的蛋白点,并对其进行MAL-DI-TOF-MS分析、鉴定,相应蛋白数据库信息检索和功能分析。Western Blot对其中2种差异蛋白进行验证。结果在BMSCs和RPE细胞鉴定出31个差异表达蛋白,其中BMSCs细胞有23种蛋白表达明显上调(主要包括SPARC、PCNA、HSP27),RPE细胞有8种蛋白表达明显上调(主要包括TBP),建立了BMSCs与RPE细胞的差异表达蛋白谱。Western Blot实验进一步验证了蛋白质组学结果。结论 SPARC、PCNA、HSP27、TBP蛋白在BMSCs和RPE细胞中具有明显表达差异,为进一步研究BMSCs诱导分化为RPE细胞提供了基础数据。

Objective To identify the difference between bone marrow （BMSCs） and retinal pigment epithelium （RPE） by the means of proteomics. mesenchymal stem cells Methods 2D-DIGE was conducted to detect different protein expression between BMSCs and RPE. The protein spots which indi- cate the difference between BMSCs and RPE were identified via MALDI-TOF-MS analysis. The protein functions were retrieved through Uniprot database. Western blot experiments were carried out on the tWO kinds of differences in protein. Results Fifty differential protein spots were identified between BMSCs and RPE. 23 protein spots were over-expressed in BMSCs comparing with RPE （including SPARC, PCNA, HSP27）, while 8 spots were down-expressed（including TBP）. Therefore, the different protein profiles between BMSCs and RPE were developed. Western blot analysis further confirms the ex- pression of some of the identified proteins. Conclusion Significantly different protein expression of SPARC, PCNA, HSP27, TBP were identified between BMSCs and RPE, which provide preliminary data for induction of BMSCs into RPE.