Proteomic Analysis of Human Intestinal Epithelial Cells

细胞生物学的一个主要挑战是确定细胞内蛋白质的亚细胞分布，并表征蛋白质定位在不同细胞生长条件下以及对应激和其他外部信号的反应如何变化。蛋白质定位通常通过显微镜或细胞分离结合蛋白质印迹技术来确定。这两种方法本质上都是低通量的，并且仅限于对已知成分的分析。我们的目标是建立基于蛋白质组学的方法来表征不同细胞系之间蛋白质表达和亚细胞定位的差异，并将这种表征扩展到人体组织。这些方法将为进一步从正常和疾病中识别潜在的生物标志物和研究组织中细胞的正常功能的相应人体组织的蛋白质组学特征提供基础。更具体地说，我们将使用这种方法来比较正常的肠上皮细胞和不同的细胞系和组织。我们的假设是，蛋白质表达和定位分析将使我们能够识别参与细胞转化和分化的蛋白质特异性组织。从这种新方法中获得的信息应用于肠道细胞和组织的研究，将产生大量的数据，识别参与分化和转化过程的蛋白质，这些数据将发展成新的假设。蛋白质组学研究领域为新颖和创新的发现提供了强大的潜力，并且在学术和工业研究实验室中也是非常受欢迎的专业知识。从长远来看，该结果将作为更详细地研究肠细胞分化和转化机制的起点。

A major challenge in cell biology is to identify the subcellular distribution of proteins within cells, and to characterize how protein localization changes under different cell growth conditions and in response to stress and other external signals. Protein localization is usually determined either by microscopy, or using cell fractionation combined with protein blotting techniques. Both these approaches are intrinsically low throughput and limited to the analysis of known components. Our objective is to establish proteomic based methods to characterize the differences in protein expression and subcellular localisation between different cell lines and extend this characterization to human tissues. These approaches will provide a basis for further proteomic characterisation of corresponding human tissues from normal and diseases to identify potential biomarkers and study normal functions of cells in tissues. More specifically, we will use this method to compare normal epithelial intestinal cells and different cell lines and tissues. Our hypothesis is that protein expression and localization profiling will allow us to identify proteins involved in cell transformation and differentiation specific to these tissues.The information gained from this new method applied to the study of intestinal cells and tissues herein will generate a large amount of data identifying proteins involved in the differentiation and transformation process that will develop into new hypotheses. The field of proteomic research presents strong potential for novel and innovative discoveries, and is also a very sought after expertise in both academic and industry research laboratories. In the long term, the results will be used as a starting point to study in more details the mechanism of differentiation and transformation of intestinal cells.