Secretome Research Of Several Kinds Of Important Cells

Secretory protein is involved in cell signal transduction, cell proliferation, differentiation/apoptosis, the organism's development and other important life processes. The understanding of the secretory protein expression characteristics of different growth period and different physiological or pathological conditions is of great significance. Secreted proteome research will not only help us to find the specific physiological /pathological condition related to biological molecules directly, and understanding of disease occurrence of molecular level or provide a theoretical basis for the development process, but also for the design of the role of specific target molecules and lay the foundation for drug discovery. This study selected several different types of primary cells and cell lines to study the model material, proteomics technology was used to study the secreted protein, it also carried out a secretory protein group methodology, expression profiling, as well as specific biomarkers of related research.First of all, we carry out a large-scale proteome research as a prerequisite for high-resolution protein separation techniques and high-throughput, high sensitivity identification technology. Therefore, in the second chapter of this study, we optimized the narrow range pH gradient two-dimensional gel electrophoresis (2-DE) separation conditions, and the use of the parameters being optimized to establish a nasopharyngeal secretion of 2-DE protein expression profiling. We then explored the in-gel protein identification strategy, which established based on high-throughput 2-DE, automatic cutting and automatic in-gel digestion of gel protein spots identification technology.Secondly, we take the advantage of the analysis of nasopharyngeal secreted proteins purification and enrichment strategy in the second chapter, and launched the olfactory ensheathing cells secreted protein expression profile studies. The difference of this chapter is that we use an optimized one-dimensional gel electrophoresis and liquid chromatography tandem mass spectrometry combined protein mixture identification technology. Olfactory ensheathing cells are a special type of glial cell which have characteristics of both astrocytes and Schwann cells. They express a high level of growth factors which play a very important role as neuronal support. In this study, ID-shotgun proteomics followed with gene ontology analysis was used to screen proteins present in the conditioned medium from primary culture rat olfactory ensheathing cells. One hundred and sixty eight non-redundant proteins were identified, the majority of which were never before reported to be produced by olfactory ensheathing cells. Furthermore, extracellular proteins were classified based on their biological and molecular functions, most of which were involved in neuronal differentiation and development. Moreover, we carried out semi-quantitative proteomic analysis to identify the active components of the olfactory ensheathing cells secretome that different from astrocytes. Spectral counting analysis revealed that twenty-three proteins were up-regulated in olfactory ensheathing cells whereas fourteen were up-regulated in astrocytes. Changes of the expression level of secreted proteins were further confirmed by western blot. In conclusion, this study provided a large number of proteins involved in neuronal development and regeneration in the olfactory ensheathing cell secretome and it may shed light on the role of extracellular matrix proteins from conditioned medium in the repairing process of central nervous system.In total, through the research papers, we have established a large-scale human nasopharyngeal carcinoma cell line secreted proteome expression profile, as well as olfactory ensheathing cells and astrocytes secreted proteome expression profile. As far as we know, our olfactory ensheathing cells secreted protein expression profile data is now the largest international database of olfactory ensheathing cells secreted protein. Moreover, this study of different strategies starting from the proteomics research has established a set of effective technology based on 2-DE and 1-DE gel electrophoresis and mass spectrometry used in conjunction with the secretion proteome research strategy can be effective for the different types of cells to secrete the protein group enrichment and simplify the analysis system, and realize potential markers of disease discovered. This secreted proteins research system will be equally applicable to other tumors or disease-related protein markers.