Abstract
The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.
Highlights
The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas
The growth of gliomas largely depends on their blood supply, the elimination of which would result in the destruction of these tumors [4]
We identified glioma-related proteins by using two-dimensional PAGE followed by MALDI-TOF-MS analysis
Summary
The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The samples were fractionated by nano-LC prior to MALDI-TOF/TOF This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. The use of laser microdissection has become an important tool in biological research to isolate relatively pure cell populations from heterogeneous frozen tissue samples [14] This technique became widely used for tracing genotypical aberrations including aberrant RNA and protein expression of subsets of cells and tissues [15]. Primary brain tumors have been subjected to direct tissue profiling and imaging mass spectrometry techniques [1, 16], to the best of our knowledge, laser microdissection of brain blood vessels has never been used before in proteomics analysis
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