Functional Gene Analysis Reveals Cell Cycle Changes and Inflammation in Endothelial Cells Irradiated with a Single X-ray Dose.
Functional Gene Analysis Reveals Cell Cycle Changes and Inflammation in Endothelial Cells Irradiated with a Single X-ray Dose.
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DOI：
10.3389/fphar.2017.00213
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发表时间：
2017
影响因子：
5.6
通讯作者：
Aerts A
中科院分区：
文献类型：
--
作者：
Baselet B;Belmans N;Coninx E;Lowe D;Janssen A;Michaux A;Tabury K;Raj K;Quintens R;Benotmane MA;Baatout S;Sonveaux P;Aerts A
背景和目的：流行病学数据表明，在低剂量（0.05和0.1戈伊）电离辐射下，患心血管疾病的风险过高。此外，辐射诱导CVD的潜在生物学和分子机制仍不清楚。由于内皮损伤在IR相关CVD中可能是关键的，因此本研究旨在确定动脉粥样硬化背景下辐射对永生化内皮细胞的影响。材料和方法：使用微阵列和RT-qPCR比较在不同的照射后（修复）时间（1天、7天、14天）测量的用单次X射线剂量（0.05、0.1、0.5、2戈伊）照射的内皮细胞的反应。为了巩固和机械地支持通过微阵列分析鉴定的内皮细胞对X射线暴露的响应，评估了DNA修复信号传导（γ H2 AX/TP 53 BP 1-病灶定量）、细胞周期进展（BrdU/7AAD流式细胞术分析）、细胞衰老（β-半乳糖苷酶测定，CPRG和IGFBP 7定量）和促炎状态（IL 6和CCL 2）。结果：微阵列结果表明细胞周期进程和炎症的持续变化。高剂量（0.5和2戈伊）后，细胞以剂量依赖性方式经历G1期阻滞，1周后通过增殖增加进行补偿，2周后几乎正常化。然而，在这一点上，照射的细胞显示出增加的β-Gal活性和IGFBP 7分泌，指示过早衰老。促炎细胞因子IL 6和CCL 2的产生在早期时间点增加。结论：IR以剂量依赖性方式诱导内皮细胞的促动脉粥样硬化过程。这些研究结果为进一步研究剂量-反应曲线的形状提供了动力，因为我们表明即使是低剂量的IR也可以在以后的时间点诱导过早的内皮衰老。此外，我们的研究结果的时间和剂量依赖性反应的差异表达的基因，细胞周期的进展，炎症和衰老带来了新的见解的基础分子机制的内皮细胞对X射线辐射的反应。这可能反过来导致风险降低策略的发展，以防止IR诱导的CVD，如使用细胞周期调节剂和抗炎药物作为辐射保护剂和/或辐射缓解剂。
Background and Purpose: Epidemiological data suggests an excess risk of cardiovascular disease (CVD) at low doses (0.05 and 0.1 Gy) of ionizing radiation (IR). Furthermore, the underlying biological and molecular mechanisms of radiation-induced CVD are still unclear. Because damage to the endothelium could be critical in IR-related CVD, this study aimed to identify the effects of radiation on immortalized endothelial cells in the context of atherosclerosis. Material and Methods: Microarrays and RT-qPCR were used to compare the response of endothelial cells irradiated with a single X-ray dose (0.05, 0.1, 0.5, 2 Gy) measured after various post-irradiation (repair) times (1 day, 7 days, 14 days). To consolidate and mechanistically support the endothelial cell response to X-ray exposure identified via microarray analysis, DNA repair signaling (γH2AX/TP53BP1-foci quantification), cell cycle progression (BrdU/7AAD flow cytometric analysis), cellular senescence (β-galactosidase assay with CPRG and IGFBP7 quantification) and pro-inflammatory status (IL6 and CCL2) was assessed. Results: Microarray results indicated persistent changes in cell cycle progression and inflammation. Cells underwent G1 arrest in a dose-dependent manner after high doses (0.5 and 2 Gy), which was compensated by increased proliferation after 1 week and almost normalized after 2 weeks. However, at this point irradiated cells showed an increased β-Gal activity and IGFBP7 secretion, indicative of premature senescence. The production of pro-inflammatory cytokines IL6 and CCL2 was increased at early time points. Conclusions: IR induces pro-atherosclerotic processes in endothelial cells in a dose-dependent manner. These findings give an incentive for further research on the shape of the dose-response curve, as we show that even low doses of IR can induce premature endothelial senescence at later time points. Furthermore, our findings on the time- and dose-dependent response regarding differentially expressed genes, cell cycle progression, inflammation and senescence bring novel insights into the underlying molecular mechanisms of the endothelial response to X-ray radiation. This may in turn lead to the development of risk-reducing strategies to prevent IR-induced CVD, such as the use of cell cycle modulators and anti-inflammatory drugs as radioprotectors and/or radiation mitigators.