title: Proteomic analysis by SILAC and 2D-DIGE reveals radiation-induced endothelial response: four key pathways.
abstract: Epidemiological data show that ionising radiation increases the risk of cardiovascular disease. The endothelium is one of the main targets of radiation-induced damage. Rapid radiation-induced alterations in the biological processes were investigated after exposure to a clinically relevant radiation dose (2.5 Gy gamma radiation). The changes in protein expression were determined using the human endothelial cell line EA.hy926 as a model. Two complementary proteomic approaches, SILAC (Stable Isotope Labelling with Amino acids in Cell culture) and 2D-DIGE (Two Dimensional Difference-in-Gel-Electrophoresis) were used. The proteomes of the endothelial cells were analysed 4h and 24h after irradiation. Differentially expressed proteins were identified and quantified by MALDI-TOF/TOF and LTQ Orbitrap tandem mass spectrometry. The deregulated proteins were mainly categorised in four key pathways: (i) glycolysis/gluconeogenesis and synthesis/degradation of ketone bodies, (ii) oxidative phosphorylation, (iii) Rho-mediated cell motility and (iv) non-homologous end joining. We suggest that these alterations facilitate the repair processes needed to overcome the stress caused by irradiation and are indicative of the vascular damage leading to radiation-induced cardio- and cerebrovascular impairment.
authors.affiliation: Helmholtz Zentrum München, German Research Center for Environmental Health, Research Unit of Radiation Cytogenetics, Neuherberg, Germany.
authors.forename: Arundhathi
authors.initials: A
authors.lastname: Sriharshan
authors.forename: Karsten
authors.initials: K
authors.lastname: Boldt
authors.forename: Hakan
authors.initials: H
authors.lastname: Sarioglu
authors.forename: Zarko
authors.initials: Z
authors.lastname: Barjaktarovic
authors.forename: Omid
authors.initials: O
authors.lastname: Azimzadeh
authors.forename: Ludwig
authors.initials: L
authors.lastname: Hieber
authors.forename: Horst
authors.initials: H
authors.lastname: Zitzelsberger
authors.forename: Marius
authors.initials: M
authors.lastname: Ueffing
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Atkinson
authors.forename: Soile
authors.initials: S
authors.lastname: Tapio
mesh_terms: D000596:Amino Acids; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000528:radiation effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*; D061307:Human Umbilical Vein Endothelial Cells / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000502:physiology / Q000528:radiation effects; D008954:Models, Biological; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D011832:Radiation Injuries / Q000378:metabolism; D015398:Signal Transduction / Q000528:radiation effects; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*; D054928:Validation Studies as Topic
chemical_list: D000596:Amino Acids; D020543:Proteome