Department of Safety and Radiation Protection, Research Centre Jülich, Germany.
Int J Radiat Biol. 2010 Oct;86(10):888-904. doi: 10.3109/09553002.2010.486016.
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.
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).
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]).
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.
定量评估人外周血淋巴细胞经γ射线照射后的早期反应蛋白（ERPRO）和早期反应基因（ERG）；鉴定特定蛋白和基因作为新型生物剂量计开发的候选生物标志物。
将人外周血淋巴细胞离体暴露于临床相关剂量（1、2 和 4 Gy）的γ射线。照射后 2 h 进行蛋白质和基因表达调控分析。使用来自一位人类供体的样本进行二维聚丙烯酰胺凝胶电泳（2D-PAGE）和 DNA 微阵列分析，以监测整体调控。在蛋白质组水平上，考虑了磷酸化和非磷酸化蛋白。使用来自多个人类供体（n=3）的样本，通过 Western blot（WB）和实时定量聚合酶链反应（RT-qPCR）技术进一步靶向感兴趣的蛋白质和基因。
在人淋巴细胞中鉴定出一组在 γ 射线照射后 2 h 显示出明显变化的 ERPRO 和 ERG。最具辐射反应性的基因和蛋白质表明细胞结构发生改变（β-肌动蛋白、talin-1[TLN1]、talin-2、zyxin-2）、免疫和防御反应（主要组织相容性复合物结合蛋白-2[MBP2]、白细胞介素-17E 和干扰素-γ）、细胞周期控制（细胞周期蛋白依赖性激酶抑制剂-1A[CDKN1A]、双微体 2、膜联蛋白 A6[ANXA6]、生长停滞和 DNA 损伤诱导蛋白-α[GADD45A]、增殖细胞核抗原[PCNA]、双重特异性磷酸酶-2 和 8[DUSP8]）以及解毒过程（过氧化物酶-1）和细胞凋亡（B 细胞淋巴瘤-2 结合成分-3[BBC3]）。
TLN1 和 CDKN1A 蛋白浓度调节的估计、ANXA6 的磷酸化状态（剂量范围 0-2 Gy）以及 MBP2 以及 BBC3、DUSP8、GADD45A 和 PCNA 的基因表达水平的改变似乎具有未来生物剂量计应用的潜力。
