Proteomic Anlaysis Of Spinal Ependymoma

Part I Obtaining and identification discrepant proteins between normal ependyma and ependymoma of spinal cord by using 2-DE proteomics technologyObjective:To evaluate the differences of proteins extracted between ependymoma and normal ependyma of spinal cord by using 2-DE combined with MALDI-TOF-MS proteomics technology. To provide a foundation for the further research to the tumorgenesis of spinal ependymoma.Method:Seven spinal ependymoma tissues were obtained through operations, four nomal ependyma tissues were obtained through autopsy The samples were progressed total proteins extraction, two-dimension electrophoresis(2-DE), quickly Coomassie brilliant blue staining, discrepant protein spots selection, spot cutting, protein enzymolysis and MALDI-TOF-MS. Then differential expression proteins were obtained and some of them were verified by Western Blotting.Results:The 2-DE gels of eleven samples from ependymoma and control groups with high resolution and reproducibility had been set up using 2-DE combined with MALDI-TOF-MS proteomics technology. Fifty effective discrepant protein spots were selected. Of the total fifty proteins, twenty-four protein spots were up-regulated and thirteen proteins spots were down-regulated in tumor group compared with the control group. Total effective discrepant protein spots were carried out by MALDI-TOF MS and MASCOT database searching Western Blotting test verified that vimentin, annexin I and HSPA8 were up-regulated in tumor group compared with the control group.Conclusion:2-DE combined with MALDI-TOF-MS proteomics technology can select discrepant protein between spinal ependymoma and normal ependyma tissues. Bioinformatical analysis indicated that vimentin, annexin I, HSPA8, calnexin and calreticulin probably related to origin of spinal ependymoma. There were no reports about these before.Partâ…¡Study on proteomic profile of normal ependyma and ependymoma of spinal cord by using SDS-PAGE-SCX-RPLC-MS/MS technologyObjective:To research focused on proteomic profile of normal ependyma and ependymoma of spinal cord by using SDS-PAGE-SCX-RPLC-MS/MS technology.Method:Seven spinal ependymoma and four nomal ependyma tissues were obtained through operations. The samples were progressed total proteins extraction, SDS-PAGE electrophoresis, gels cutting, protein enzymolysis and SCX-RPLC-LTQ-Orbitrap. The samples were progressed total proteins extraction. The identified proteins were analysis by bioinformatics with DAVID (The Database for Annotation, Visualization and Integrated Discovery) and KEGG (Kyoto Encyclopedia of Genes and Genomes).Results:We established three-diamensional SDS-PAGE-SCX-RPLC-MS/MS technology. A normal spinal ependyma proteome consist of 2758 proteins while the ependymoma proteome consist of 2721.Conclusion:It is the first time in world to proteomic profile of normal ependyma and ependymoma of spinal cord by using SDS-PAGE-SCX-RPLC-MS/MS technology. MAPK signal pathway probably correlated with the spinal ependymoma All the proteomic profile studies provide a foundation for the further tumor-genetical research of spinal ependymoma.