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目的寻找不同分化潜能的真皮干细胞的差异膜蛋白。方法通过选择性地抽提三向分化克隆的细胞膜蛋白,与二向分化克隆进行对比分析,以筛选出与分化能力下降有关的细胞膜相关蛋白。三向克隆与双向克隆的膜蛋白经2-D电泳、质谱分析及Western-Blot检测,初步确认二者之间的明显差异蛋白。结果蛋白质组学分析结果显示,二者的差异蛋白主要有10种膜相关蛋白,其中膜表达差异较显著的蛋白质,如:Annexin A2 isoform1(ANXA2)、Prohibitin(PHB)、Protein disulfide-isomeraseA3(PDIA3),均与细胞分化有关。结论ANXA2、PHB、PDIA3在不同分化潜能的人真皮干细胞膜上表达存在差异,提示了这几组蛋白质参与了该细胞的分化,为初步筛选多潜能真皮成纤维细胞,以及进一步研究其分化的机理提供了重要科学依据和深入探索的新方向。
Objective To find differential membrane proteins of dermal stem cells with different differentiation potential. Methods The cell membrane proteins of three-way differentiated clones were selectively extracted and compared with the two-way differentiated clones to screen the cell membrane-associated proteins related to the diminished ability of differentiation. Three-way cloning and two-way cloning of membrane proteins by 2-D electrophoresis, mass spectrometry and Western-Blot detection, initially identified significant differences between the two proteins. Results The results of proteomic analysis showed that there were 10 kinds of membrane-associated proteins in the differential proteins, including proteins with significant differences in membrane expression such as Annexin A2 isoform1 (ANXA2), Prohibitin (PHB), Protein disulfide-isomeraseA3 ), Are related to cell differentiation. Conclusion The expression of ANXA2, PHB and PDIA3 in human dermal stem cell membranes with different differentiation potentials is different, suggesting that these groups of proteins are involved in the differentiation of the cells. In order to preliminary screen the pluripotent dermal fibroblasts and to further study its differentiation mechanism Provide important scientific basis and explore new directions in new directions.