STOREDB:STUDY1063 Proteome analysis of irradiated endothelial cells reveals persistent alteration of RhoGDI and NO signalling pathways [DOI:10.20348/STOREDB/1063]
|Proteome analysis of irradiated endothelial cells reveals persistent alteration of RhoGDI and NO signalling pathways|
|Published: Open access to everyone|
|DATA SHARING POLICY|
|CC-Attribution Non-Commercial Share Alike|
|SIZE OF COHORT|
|MELODI RESEARCH PRIORITY|
|Analysis of mechanisms involved in low dose radiation through use and development of suitable cellular and animal models|
|INTERNAL OR EXTERNAL EXPOSURE|
|TYPE OF EXTERNAL EXPOSURE|
|BIOLOGICAL SAMPLE AVAILABLE|
|Background: Recent epidemiological studies indicate that radiation doses as low as 0.5 Gy may increase the risk of cardiovascular disease. The aim of the present study was to investigate whether this radiation dose causes molecular alterations in endothelial cells that could support the suspicion of low-dose cardiovascular risk raised by population studies.
Materials and Methods: Radiation-induced changes in the proteome of human coronary artery endothelial cell line were analysed using triplex isotope-coded protein labelling technology combined with LC–ESI–MS/MS. The cells were irradiated using the dose of 0.5 Gy (X-ray) and investigated after different time intervals (1 day, 1 week, 2 weeks). The proteomics and bioinformatics data were validated by immunoblotting and ELISA.
Results: The radiation-induced alteration of the endothelial proteome was characterised by perturbation of RhoGDI and NO signalling pathways. This was accompanied with reduced proteasome activity and enhanced protein carbonylation indicating augmented oxidative stress.
Conclusions: The persistent adverse molecular changes found in this study are indicative of endothelial dysfunction and support the epidemiological data showing significantly increased risk of cardiovascular disease at 0.5 Gy.
|MEAN DURATION OF FOLLOW-UP (WEEKS)|
STOREDB:DATASET1102 EC 500 mGy 1 day, 7 days and 14 days [DOI:10.20348/STOREDB/1063/1102]
Created on:2017-01-11 11:15:40 Modified On:2017-01-11 11:15:40
|EC 500 mGy 1 day, 7 days and 14 days|
|ICPL labelling and analysis was done as previously reported . Briefly, triplicate aliquots of 50 µg protein lysate obtained from either control or irradiated samples were labelled with Isotope Coded Protein Label (ICPL) reagents (SERVA) following the manufacturer’s instructions. ICPL0 was used for the control samples and ICPL6 for the irradiated samples. Samples representing the three different time points after irradiation were analysed independently. The heavy and light labelled samples were combined and then separated by 12% SDS gel electrophoresis before mass spectrometry analysis using nano-LC connected to a linear quadrupole ion trap- Orbitrap mass spectrometer (LTQ Orbitrap XL, ThermoFisher, Bremen, Germany) and equipped with a nano-ESI source as previously described [27, 28].
Data processing for protein quantification of ICPL pairs was performed using MASCOT search engine (version 2.3.02; Matrix Science) and Proteome Discoverer version 1.3 (Thermo Fisher) as described in detail previously .