PuSH - Publication Server of Helmholtz Zentrum München: Proteomic analysis by SILAC and 2D-DIGE reveals radiation-induced endothelial response: Four key pathways.

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Sriharshan, A. ; Boldt, K. ; Sarioglu, H. ; Barjaktarovic, Z. ; Azimzadeh, O. ; Hieber, L. ; Zitzelsberger, H. ; Ueffing, M. ; Atkinson, M.J. ; Tapio, S.

Proteomic analysis by SILAC and 2D-DIGE reveals radiation-induced endothelial response: Four key pathways.

J. Proteomics 75, 2319-2330 (2012)

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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 (2.5Gy 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.

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