Analysis Of Plasma Differential Protein In Brain Glioblastoma

Background:Glioma is the most commonly seen malignant tumor in nervous system. It derived fromneuroglia cell which originated in neuroectoderm and account for80%of the malignant braintumor. It was found that the more malignant of the tumor, the lower of the cure rate for the patients.The mean survival time of glioblastoma in grade Ⅳ was only12-15months, which was greatlydifferent from other kind of tumor (usually with5-10years of median survival). Besides, it is easyfor glioma to recur, and once the recurrence was happen, it often accompanied with grade elevation,sensitivity reduction to radiotherapy/chemotherapy, and drug resistance.The diagnostic of tumor are mainly based on imaging and histopathology method.Nevertheless, due to glioma’s character in complexity, diversity and untypicality, imaging methodcould not determine the category and grde of glioma, despite its advantage in visual sense. On theother hand, though histopathology was wide used in tumor category determination, its applicationin early diagnosis and real time therapy monitoring was greatly hindered for its invasive nature.Meanwhile, it’s well known that the judgment of histopathology is subjective, and diverse resultsmay be achieved for a same sample among different pathologist. If glioma specific protein couldbe explored from the peripheral blood of the patients, then more information of the patients inprognosis, therapy evaluation and recurrence monitor may be obtained, which may help thephysicians to fulfill better the individualized treatment to the patients.Although blood sample is easy to obtain, the separation and identification of the plasmaprotein is hard for traditional methods. However, mass spectrum has obvious advantages in thisarea, and it was reported that more than2000kinds of proteins and polypeptides can be separatedby NanoUPLC-nano-ESI-MS/MS, which making it possible to screen out potential glioma specificprotein.Purpose:In this study, applied comparative proteomic technology, through the separation、analysisand comparison human plasma proteome of primary brain glioblastoma patients and the healthy,find specific plasma protein markers of brain glioblastoma. It will provide meaningful assistance toclarify the pathogenesis of brain glioblastoma， screening and research of targeted therapeuticdrugs, as well as the early diagnosis, prognosis, treatment options, efficacy evaluation, detection ofrecurrence of brain glioblastoma patients Method:Randomly selected six cases from all primary brain glioblastoma patients and healthy fastingplasma specimens. First Bradford applied for measured the protein content of the sample, andremove non-protein components. Then using two-dimensional gel electrophoresis(2DE),differentially expressed proteins were isolated after gel silver staining and gel imagerscaned images using BIO-RAD GS-800. The obtained images has analyzed the differences withPDQuest Advanced-8.0.1analysis software between more than two-fold difference in points. Afterin-gel digestion, It applied with the ultra-efficient nanoliter liquid chromatography-tandem massspectrometry (NanoUPLC-nano-ESI-MS/MS) isolation and identification technology. Afterprocessing with the software PLGS v2.3, using Mascot software, according to the MS/MS ionmode, retrieve IPI (International Protein Index) Database database. After a comparative analysis ofdata screening and identification of differentially expressed proteins.Results:Two plasma proteins separated by2DE, through software analysis, differences in expressionlevels were screened out the four differentially expressed proteins more than twice. AfterNanoUPLC-nano-ESI-MS/MS mass isolated and identified, Mascot software analysis database, get23different upregulated.proteins．To include of two unnamed protein products, Human serumalbuminmutant R218h complexed with thyroxine, Human serum albumic omplexed with Myristateand Azapropazone, Human serum albumin complexed with Myristate and Aspirin, ApolipoproteinD,Platelet basic protein preproprotein,Thrombocidin-2antimicrobial variant,Dermcidinpreproprotein, Structure of quaternary complex of human Tlr3ecd with threefabs,IGL@protein,protein NIG58, Bence-Jones, Anti-Il-15antibody in complex with humanIl-15,Anti-peptid/MHCcomplex HLA-A1/MAGE-A1mono-clonal antibody light chain，Crystal structure of a diseasessociate anti-human Gm-Csf autoantibody, GAD65-specific monoclonal antibody b78lambdachain,Protein S100-A7,Protein S100-A8,Lysozyme precursor, Mutant human lysozyme C77a,Human lysozyme double mutant A96l、W109h,Complement component C3, Proapolipoprotein．Conclusion:First application2DE and NanoUPLC-nano-ESI-MS/MS technical analysis of brainglioblastoma plasma proteome. The total screened23differential proteins in glioblastoma plasma.and not in the healthy people. All of them may be involved in the regulation process ofglioblastoma. Combined with a variety of different proteins may constitute a specific diagnosticmodel of glioblastoma. For early diagnosis, the prognosis, treatment options, efficacy evaluation,monitoring and other aspects of recurrence of glioblastoma provide useful help.