STOREDB:STUDY1063 Proteome analysis of irradiated endothelial cells reveals persistent alteration of RhoGDI and NO signalling pathways [DOI:10.20348/STOREDB/1063]

Study meta-data


STUDYIDSTOREDB:STUDY1063
CREATEDON2017-01-11 11:14:31
MODIFIEDON2017-01-11 11:14:31
UPLOADEROmid Azimzadeh
DOIDOI:10.20348/STOREDB/1063

Study details


STUDY NAME
Proteome analysis of irradiated endothelial cells reveals persistent alteration of RhoGDI and NO signalling pathways
STUDY STATUS
Published: Open access to everyone
DATA SHARING POLICY
CC-Attribution Non-Commercial Share Alike
COUNTRY
Germany
PRINCIPAL INVESTIGATOR
Omid Azimzadeh
SPECIES
Homo sapiens
SIZE OF COHORT
0-999
OUTCOME
Cardiovascular
MELODI RESEARCH PRIORITY
Analysis of mechanisms involved in low dose radiation through use and development of suitable cellular and animal models
EXPOSURE CONTEXT
Medical
INTERNAL OR EXTERNAL EXPOSURE
External
TYPE OF EXTERNAL EXPOSURE
X-ray
EXPOSURE PATTERN
Acute
DOSE RATE
Low
BIOLOGICAL SAMPLE AVAILABLE
Yes
STUDY DESCRIPTION
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.
PUBMED ID
1
MEAN DURATION OF FOLLOW-UP (WEEKS)
14

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
DATASET NAME
EC 500 mGy 1 day, 7 days and 14 days
DOIDOI:10.20348/STOREDB/1063/1102
DATASET DESCRIPTION
ICPL labelling and analysis was done as previously reported [26]. 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 [26].