Proteomics Study Of The Colorectal Cancer In Dukes B Period

The Colorectal cancer is one of current global most common mali-gnant tumors, its incidence rate and the mortality rate is increasing year by year. The Colorectal cancer tended to shift and recur easily, and has become the5th in mortality rate of maligrant tumor in our country, it is quite necessary to conduct the research of its morbidity, the shift and the recrudescence mechanism .Traditionally, the domestic and foreign scholars,researches of colorectal cancer mainly focus on :â‘ Epidemiology investigation, foresightedness;â‘¡Foresighted and retrospective comparative studiesâ‘¢autopsy analysis. Nowadays, along with scientific progress, the methods for the study of colon cancer are increasing rapidly, and the studying fields are more and more broad. Such as histochemistry dyes, PCR method, protein studies and so on. Particularly in recent years, the research in Protein levels developed more quickly,and the study on colorectal cancer has steped into a new world.With the human genome project's implementation and advancement,life science research has entered the post-genome era. In this era, the main studying object of life science is functional genomics, including structural genomics and proteomics research.Although there are many species' genome has been sequenced, but more than half of gene's function is unknown in these genomes . Currently, the strategies of the functional genomics uses, such as gene chip, serial analysis of gene expression (SAGE), which are considered in the context of mRNA from the cells , and its premise is that the level of mRNA of cells can reflect the protein expression levels. But this is not entirely the case, in the mRNA point of view, actually, it only includes the level of transcription regulation, and can not fully represent the level of protein expression. Protein's complex post-translational modification, protein's sub-cellular localization or migration, and protein-protein interactions is almost impossible to judge from the mRNA level.Needless to say, protein is executor of physiological functions, is the direct embodiment of the phenomenon of life, and the study of protein structure and function will directly clarify the conditions of life under physiological or pathological changes in the mechanism. The existence forms and activity patterns of the protein itself, such as post-translational modification, protein-protein interactions and protein conformation and so on, still depends on the direct study of proteins to resolve. Although the protein's special nature, such as variability, diversity and so on, has led to the result that the research techniques of protein are far more complex and difficult than nucleic acid technology , but it is these characteristics affect the entire life course.The traditional study approach of a single protein has been unable to meet the requirements of post-genome era, it is because [2]:â‘ Biological phenomena's occurrence is often multi-factors,it will inevitably involve multiple proteins;â‘¡the involvement of multiple proteins is interwoven into a network, or parallel to occur, or showed a cascade relationship;â‘¢In the implementation of physiological functions, the performance of proteins are diverse, dynamic, it is not as basically fixed as the genome. Therefore,if one need to have a comprehensive and in-depth understanding of the complexity of life, it is essential to study protein on a overall, dynamic, network level. Thus, in 1994, Australia's Macquarie University, Wilkins and Williams first proposed proteome concept, which originated from the heterozygosity of two words,protein and genome , which is defined as: all types of proteins expressed by a cell in a particular physiological or pathological state. Proteomics which takes all proteins expressed by the genome, namely protein group,as studying object, is a new area of research. Proteomics is divided into two types: 1,expressional proteomics which is to study the change of quantity of protein expression between different samples .2,functional proteomics which establish cell signal transduction pathway network through the separation of protein complexes and the systematic study protein-protein interactions . Proteomics has a rich studying content ,a great many technology platform and more and more advanced technological means. In that case, rational selection of clinical specimens, is the key to the expressional proteomics studies.Generally, International organizations still follow the Dukes improved TNM staging and the staging method proposed by UICC. According to the Dukes law's supplementin our country , divides into: Cancers limited in the intestinal wall is within the DukesA period. Serosal invasion through the intestinal wall and / or serosa things, but no lymph node metastasis is the B phase. With lymph node metastasis belongs to the C phase, in which lymph node metastasis is limited to the vicinity, such as colon wall and lymph nodes are for the C1 period; if transferred to the Department of Film and mesangial root node is the C2 period. Already have distant metastasis or peritoneal metastasis or extensive invasion to adjacent organs and can not be removed is the D periodBased on the above principle, this experiment identified stages of clinical pathology, the Patient tumor organization of Colorectal cancer in Dukes B period and different proteins of adjacent tissues in the pathological classification through the proteomic approach.,it can provide protein markers for the pathological classification of colorectal cancer ;it can be further used to find new targets for early diagnosis and biology treatment of colorectal cancer .Under the application of two-dimensional gel electrophoresis and mass spectrometry experiments , we finally obtained organization different proteins of tumors and adjacent tissues 21, including 14 up- regulated proteins and 7 protein down. With the application of two-dimensional LC-MS analysis, we could obtain 44 organization different proteins of cancer and adjacent tissues, including 21 up- regulated proteins and 23 protein down. From the two technical route we find 13 proteins to obtain the consistent appraisal result, and including 7 up- regulated proteins ,for example, Hemoglobin Lepore-Baltimore, 60 kDa heat shock protein, ATP synthase subunit alpha , Keratin type I cytoskeletal 18 , Protein disulfide-isomerase A3,Alpha-enolase,Isoform 1 of Nucleophosmin;and 6 protein down, for example, Isoform 1 of Desmuslin,Transgelin,Desmin,Calponin-1,Heat shock protein beta-1,Isoform 2 of Vinculin.In order to confirm the accuracy of our experiment,we selected two protein(Enolase,Hemogobin) among the 21different proteins of cancer and adjacent tissues by Dimensional gel electrophoresis and mass spectrometry experiments respectively and verified the accuracy of two-dimensional gel electrophoresis by immunity signature experiment; in the same way , we also selected two protein(Vinculin,Talin)among 44 different proteins which were finally received from tumor and adjacentissues in the two-dimensional liquid chromatography analysis and verified the accuracy of Two-dimensional LC-MS experiment by immunity signature experiment. In addition ,we discussed the four proteins above ,which further illustrates their meaning in the incidence of disease ,colorectal cancer diagnosis or treatment and other aspects.