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P04.202-D FLUORESCENCE DIFFERENCE GEL ELECTROPHORESIS (DIGE) AND MASS SPECTROMETRY ANALYSIS IN EPENDIMOMAS: PRELIMINARY DATA

ABSTRACT: Despite the histological similarity of ependymomas from throughout the neuroaxis, the disease comprises multiple entities, depending on site of origin (spinal cord, posterior fossa and supratentorial) and patient age (children versus adults). Although detailed histological examination may identify important prognostic factors, their strength varies between subsets. Clinically, ependymomas remain an enigma to neuro-oncologists and neurosurgeons. Proteomic data on ependimomas are scant but a more detailed insight may complement cytogenetic and gene expression data in the effort to achieve a robust classification system of pediatric and adult ependymomas as a base for a molecular based therapy. RNA, DNA and proteins were extracted from 14 selected ependimomas. Proteins were analyzed with a 2-D Fluorescence Difference gel Electrophoresis (2D-DIGE) to compare proteomes. DIGE spots demonstrated significant difference in protein expression between selected types of ependimoma. Mass spectrometry analysis of these spots identified a high presence of GFAP and Albumin, Vimentin and Creatine kinase B-type. A new protein of particular interest - never described in ependimomas and other tumours of Central Nervous System - is FERM domain-containing protein 3 (FRMD3) (accession A2A2Y4). FRMD3 is as members of the 4.1 protein superfamily, which shares the highly conserved membrane-association domain FERM, which has already been linked to tumour suppressor activity. These preliminary data will be extended to more cases to discovery new biomarkers useful to integrate molecular data and possible validate diagnostic and/or prognostic factors.

SUBMITTER: Morra I

PROVIDER: S-EPMC4185497 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND: Peach fruit undergoes a rapid softening process that involves a number of metabolic changes. Storing fruit at low temperatures has been widely used to extend its postharvest life. However, this leads to undesired changes, such as mealiness and browning, which affect the quality of the fruit. In this study, a 2-D DIGE approach was designed to screen for differentially accumulated proteins in peach fruit during normal softening as well as under conditions that led to fruit chilling injury. RESULTS: The analysis allowed us to identify 43 spots -representing about 18% of the total number analyzed- that show statistically significant changes. Thirty-nine of the proteins could be identified by mass spectrometry. Some of the proteins that changed during postharvest had been related to peach fruit ripening and cold stress in the past. However, we identified other proteins that had not been linked to these processes. A graphical display of the relationship between the differentially accumulated proteins was obtained using pairwise average-linkage cluster analysis and principal component analysis. Proteins such as endopolygalacturonase, catalase, NADP-dependent isocitrate dehydrogenase, pectin methylesterase and dehydrins were found to be very important for distinguishing between healthy and chill injured fruit. A categorization of the differentially accumulated proteins was performed using Gene Ontology annotation. The results showed that the 'response to stress', 'cellular homeostasis', 'metabolism of carbohydrates' and 'amino acid metabolism' biological processes were affected the most during the postharvest. CONCLUSIONS: Using a comparative proteomic approach with 2-D DIGE allowed us to identify proteins that showed stage-specific changes in their accumulation pattern. Several proteins that are related to response to stress, cellular homeostasis, cellular component organization and carbohydrate metabolism were detected as being differentially accumulated. Finally, a significant proportion of the proteins identified had not been associated with softening, cold storage or chilling injury-altered fruit before; thus, comparative proteomics has proven to be a valuable tool for understanding fruit softening and postharvest.

Dataset Information

P04.202-D FLUORESCENCE DIFFERENCE GEL ELECTROPHORESIS (DIGE) AND MASS SPECTROMETRY ANALYSIS IN EPENDIMOMAS: PRELIMINARY DATA

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