Department of Physiology, School of Medicine, CHA University, Seongnam, Gyeonggi, Republic of Korea.
Pancreas. 2012 Jan;41(1):65-73. doi: 10.1097/MPA.0b013e3182223e35.
This study aimed to identify proteins important for the primitive gut tube differentiation from human embryonic stem cells (hESCs) by derivation method for pancreatic cells.
Proteins with altered expression levels in the process of differentiating to primitive gut tube from definitive endoderm of hESCs were investigated by comparative proteomic analysis using 2-dimensional gel electrophoresis and mass spectrometric analyses.
Differentiation to primitive gut tube from hESCs was analyzed using differentiation marker genes and proteins. Twenty-seven protein spots with significant changes in intensity were found by 2-dimensional gel electrophoresis, and 24 proteins were further identified. These proteins were functionally annotated based on gene ontology. The expression levels of 3 proteins, RREB1, PDE6B, and CD209, involved in signal transduction, were validated using quantitative reverse transcription-polymerase chain reaction and Western blot. Their mRNA and protein expression levels increased in primitive gut tube but not in definitive endoderm or embryonic body.
The increase in expression of RREB1, PDE6B, and CD209 suggests that these proteins might play important roles in the differentiation of primitive gut tube cells from hESCs and in human primitive gut tube development into pancreas. Therefore, they could be developed as differentiation markers for identifying primitive gut tube cells.
本研究旨在通过胰腺细胞衍生方法，从人类胚胎干细胞（hESC）中鉴定出对原始肠管分化重要的蛋白质。
通过比较蛋白质组学分析，使用二维凝胶电泳和质谱分析，研究了从 hESC 向原肠管分化过程中表达水平发生变化的蛋白质。
使用分化标记基因和蛋白质分析 hESC 向原始肠管的分化。通过二维凝胶电泳发现 27 个蛋白斑点的强度有明显变化，进一步鉴定出 24 种蛋白质。根据基因本体论对这些蛋白质进行了功能注释。使用定量逆转录-聚合酶链反应和 Western blot 验证了 3 种蛋白质（RREB1、PDE6B 和 CD209）的表达水平，这些蛋白质参与信号转导，在原始肠管中表达增加，但在定型内胚层或胚胎体中不表达。
RREB1、PDE6B 和 CD209 的表达增加表明，这些蛋白质可能在 hESC 原始肠管细胞的分化和人类原始肠管向胰腺的发育中发挥重要作用。因此，它们可以作为鉴定原始肠管细胞的分化标记物。
