X射线离体照射人外周血淋巴细胞差异表达基因的筛选与初步验证

摘要/Abstract

摘要： 筛选外周血淋巴细胞中的辐射应答差异表达基因(differrentially expression gene, DEGs)并验证其剂量-效应和时间-效应关系,探讨DEGs作为辐射生物剂量计的可行性。取3名成年志愿者的外周血淋巴细胞样本进行0.1、0.2、0.5、1、2和4 Gy X射线照射,对细胞进行RNA测序及生物信息学分析。以log₂ |Fold Change| ≥2、P<0.05为条件筛选DEGs并进行聚类热图分析、基因本体论(GO)功能注释分析和京都基因与基因组百科全书(KEGG)信号通路富集分析。实时荧光定量PCR(qRT-PCR)验证DEGs中EDA2R、ACER2、CD70、ACTA2、MGAT3、MAMDC4的剂量-响应或时间-效应关系。照射后 8 h在不同剂量(0.1、0.2、0.5、1、2和4 Gy)水平上筛选出37个DEGs,GO分析表明DEGs基因在细胞过程、单个生物体过程、生物调节、对刺激应答的调控中最为丰富,而KEGG分析表明DEGs主要在细胞生长和死亡过程中富集。qRT-PCR实验结果表明EDA2R、ACER2、CD70、ACTA2在照射后8 h以剂量依赖的方式显著上调。2 Gy X射线照射淋巴细胞中ACTA2的表达随时间增加而增加,表现出时间-效应关系。人外周血淋巴细胞样本中EDA2R、ACER2、CD70、ACTA2呈剂量依赖性上调,其中ACTA2呈时间依赖性上调,这些基因具有成为人类辐射生物剂量计的潜能。

关键词: 电离辐射, 外周血淋巴细胞, 差异表达基因, 剂量-响应关系

Abstract: To screen the differentially expression genes (DEGs) in peripheral blood lymphocytes and verify the dose-response and temporal response, and to explore the feasibility of DEGs as a radiation biological dosimeter. Human peripheral blood samples from 3 adult volunteers were exposed to X-rays at doses of 0, 0.1, 0.2, 0.5, 1, 2 and 4 Gy. RNA-sequencing and bioinformatics analyses were performed. DEGs were screened out under the conditions of log₂|Fold Change| ≥2,P<0.05. The cluster heat map analysis, Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were applied for the identification of DEGs. Among these DEGs, EDA2R, ACER2, CD70, ACTA2, MGAT3, MAMDC4 were verified for the dose response and temporal response to ionizing radiation by qRT-PCR. We identified 37 DEGs at doses of 0, 0.1, 0.2, 0.5, 1, 2 and 4 Gy at 8 h post-irradiation. The GO analysis of DEGs was mainly enriched in cellular process, single-organism process, cell, cell part, binding. KEGG pathway analysis highlighted the significance of cell growth and death. Doses of 0, 0.1, 0.2, 0.5, 1, 2 and 4 Gy led to a dose-dependent increase in the mRNA levels of EDA2R, ACER2, CD70, ACTA2. The mRNA expression of ACTA2 after 2 Gy X-ray irradiation was shown to be elevated in a time-dependent manner, revealed a temporal response to ionizing radiation. A dose-dependent increase in expression of EDA2R, ACER2, CD70, ACTA2,and a time-dependent increase in ACTA2 in human peripheral lymphocyte samples were demonstrated, indicating the potential of these genes to be biomarkers of human radiation exposure.

Key words: ionizing radiation, peripheral blood lymphocyte, differentially expression gene, dose-response relationship

中图分类号:

于慧杰, 宋秀军, 李峰生, 江其生, 王思念, 李伟. X射线离体照射人外周血淋巴细胞差异表达基因的筛选与初步验证[J]. 辐射防护通讯, 2026, 46(2): 19-25.

YU Huijie, SONG Xiujun, LI Fengsheng, JIANG Qisheng, WANG Sinian, LI Wei. Screening and preliminary verification of differentially expressed genes in ex vivo X-irradiated human peripheral blood lymphocytes[J]. RADIATION PROTECTION BULLETIN, 2026, 46(2): 19-25.

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