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Establishment of Two-dimension Gel Electrophoresis Protein Profiles and Differential Aanalysis from Human Pituitary Adenoma with and without Invasivness

Author: ChenWei
Tutor: LiuZhiXiong
School: Central South University
Course: Surgery
Keywords: Pituitary adenoma Invasive Differential Proteomics 2-D electrophoresis Mass Spectrometry Oncogene DJ-1 protein Heat shock protein 27 Western blot
CLC: R736.4
Type: Master's thesis
Year: 2008
Downloads: 69
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Abstract


Objective: Using a proteomics approach, for the first time to establish a credible human invasive and non-invasive pituitary adenomas two-dimensional gel electrophoresis protein profiles, to study the differential expression of the protein identification, authentication and invasiveness of pituitary adenomas lay the foundation. Materials and Methods: 20 cases of collection Xiangya Hospital neurosurgery pituitary adenoma specimens, waiting for the biopsy results to screen invasive, invasive and non-invasive pituitary adenomas 10 cases each. Purification of pituitary adenomas total protein immobilized pH gradient (IPG) two-dimensional gel electrophoresis (2-DE) separation of total protein gel by Coomassie blue staining after the application of the ImageMaster 2D image analysis software analysis to identify differentially expressed proteins using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) mass spectrometry analysis of the differentially expressed protein spots, corresponding peptide mass fingerprinting (PMF), the identification of differentially expressed proteins of the search database. Results: (1) for the first time two-dimensional gel electrophoresis protein profiles credible human invasive and non-invasive pituitary adenomas. Each of three randomly selected invasive and paired non-invasive pituitary adenomas gel average spots respectively for 1080 ± 24,1035 ± 28 the average match points to 909 ± 45,828 ± 56, average match percentage of 84.3 %, 79.5%; the protein points in the same tissue type of glue, etc. isoelectric focusing (IEF) direction of the deviation is (1.563 ± 0.259) mm, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) direction on the deviation of (1.088 ± 0.206) mm; (2) comparative analysis of two-dimensional gel electrophoresis protein profiles of human invasive and non-invasive pituitary adenoma condensate between the average match points to 899 ± 58 match percentage of 83.3%, random peptide mass fingerprint analysis identified with cell growth and proliferation, metabolism, cell cycle regulation, signal transduction protein of 30 different protein spots. Conclusion: This study for the first time established a credible invasive pituitary adenomas two-dimensional gel electrophoresis protein profiles in identification of oncogene DJ-1 protein, heat shock protein 27 and some protein may be associated with invasiveness of pituitary adenomas . Purpose: This topic aims to study pituitary adenomas oncogene DJ-1 protein (DJ-1), heat shock protein 27 (HSP27) expression whether its invasiveness, research associated with tumor close DJ-1, HSP27 the expression level as the possibility to determine the invasiveness of pituitary adenomas and prognostic indicators. MATERIALS AND METHODS: selected 20 patients with resected pathological diagnosis of pituitary adenoma, complete endocrine data, according to Hardy-Wilson and Knosp classification of staging, combined with magnetic resonance images and biopsy to determine the tumor aggressiveness, application Western blot (Western Blot) were detected in each tumor, DJ-1, HSP27 expression levels, and to analyze the relationship between DJ-1, HSP27 and tumor aggressiveness. On the results of the application of SPSS 13.0 forWindows package for statistical analysis. Results: (1): 10 cases of invasive pituitary adenomas DJ-1 protein expression of HSP27 strongly positive rates were 70% (7/10), 80% (8/10), 10 cases of non-invasive pituitary adenomas DJ-1 protein expression of HSP27 strong positive rate was 10% (1/10), 10% (1/10), invasive pituitary adenomas was significantly higher than the non-invasive pituitary adenomas (P <0.05). (2) DJ-1, there is a significant positive correlation of HSP27 in the expression level of invasiveness of pituitary adenomas. Conclusion: oncogene DJ-1 or HSP27 expression in invasive pituitary adenomas strong positive rate was significantly higher than the non-invasive pituitary adenomas (70% / 10%, 80% / 10%, P <0.05). Oncogene DJ-1, abnormal expression of HSP27 in protein plays a more important role in the formation and development of the pituitary adenoma in, suggesting that the DJ-1, of HSP27 protein expression of the strength of the pituitary adenoma invasion extent, the expression levels may as judged by the invasiveness of pituitary adenomas one of the indicators of prognosis and treatment.

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CLC: > Medicine, health > Oncology > Internal endocrine tumors > Pituitary tumor
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