Proteomic and genomic modulations induced by γ-irradiation of human blood lymphocytes.

Abstract

PURPOSE:

Quantitative evaluation of early response proteins (ERPRO) and early response genes (ERG) following γ-irradiation of human lymphocytes; identification of specific proteins and genes as candidate biomarkers for the development of a novel biodosimeter.

MATERIALS AND METHODS:

Human peripheral blood lymphocytes were exposed to clinically relevant doses (1, 2 and 4 Gy) of γ-radiation ex-vivo. Analyses of protein and gene expression modulation were conducted 2 h post-irradiation. Global modulations were monitored using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and DNA microarray analyses of the samples originating from one human donor. On the proteome level, both phosphorylated and non-phosphorylated proteins were considered. Proteins and genes of specific interest were further targeted using Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) techniques, employing samples from several human donors (n=3).

RESULTS:

A set of ERPRO and ERG showing significant alterations 2 h post-γ-irradiation have been identified in human lymphocytes. The most radiation responsive genes and proteins indicated alterations of cellular structure (ß-actin, talin-1 [TLN1], talin-2, zyxin-2), immune and defence reactions (major histocompatibility complex binding protein-2 [MBP2], interleukin-17E and interferon-γ), cell cycle control (cyclin-dependent kinase inhibitor-1A [CDKN1A], mouse double minute-2, annexin-A6 [ANXA6], growth arrest and DNA-damage-inducible protein-α [GADD45A], proliferating cell nuclear antigen [PCNA], dual specificity phosphatase-2 and 8 [DUSP8]) as well as detoxification processes (peroxin-1) and apoptosis (B-cell lymphoma-2 binding component-3 [BBC3]).

SUMMARY:

The estimations of protein concentration modulation of TLN1 and CDKN1A, phosphorylation status of ANXA6 (dose range 0-2 Gy) and MBP2 as well as the alterations in the level of gene expressions of BBC3, DUSP8, GADD45A and PCNA appears to be of potential value for future biodosimetric applications.

• Authors: Andrei Turtoi, Rajeshwar N Sharan, Alok Srivastava, Frank H A Schneeweiss
• Journal: International journal of radiation biology (Int J Radiat Biol) Vol. 86 Issue 10 Pg. 888-904 (Oct 2010) ISSN: 1362-3095 [Electronic] England
• PMID: 20653344 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Proteome

Topics

• Adult
• Blotting, Western
• Dose-Response Relationship, Radiation
• Female
• Gamma Rays
• Gene Expression Profiling
• Genome, Human (radiation effects)
• Genomics
• Humans
• Lymphocytes (metabolism, radiation effects)
• Male
• Mass Spectrometry
• Oligonucleotide Array Sequence Analysis
• Proteome (radiation effects)
• Proteomics
• Reproducibility of Results
• Reverse Transcriptase Polymerase Chain Reaction
• Time Factors