| 王亚林,李爽,孙鑫,等.低剂量辐射诱导差异表达人淋巴细胞lncRNAs的筛选与初步验证[J].中华放射医学与防护杂志,2025,45(5):423-430.Wang Yalin,Li Shuang,Sun Xin,et al.Screening and preliminary validation of differentially expressed lncRNAs in human lymphocytes induced by low dose ionizing radiation[J].Chin J Radiol Med Prot,2025,45(5):423-430 |
| 低剂量辐射诱导差异表达人淋巴细胞lncRNAs的筛选与初步验证 |
| Screening and preliminary validation of differentially expressed lncRNAs in human lymphocytes induced by low dose ionizing radiation |
| 投稿时间：2025-01-06 |
| DOI：10.3760/cma.j.cn112271-20250106-00009 |
| 中文关键词: 电离辐射 低剂量 长链非编码RNA 生物标志物 |
| 英文关键词:Ionizing radiation Low dose Long non-coding ribonucleic acid (lncRNA) Biomarker |
| 基金项目:国家自然科学基金(82173464,82173463) |
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| 中文摘要: |
| 目的 探讨人淋巴细胞长链非编码RNA (lncRNAs)在低剂量电离辐射诱导下表达水平的改变及其作为辐射生物标志物的可能性。方法 采用0、0.05和0.1 Gy γ射线照射人永生化淋巴细胞(AHH-1),照后24 h提取RNA进行全转录组测序,测序分为0、0.05和0.1 Gy 3组,筛选低剂量辐射诱导的差异表达lncRNAs;采用基因本体论(GO)分析差异表达基因的分子功能、生物学过程以及信号通路的富集情况;采用qRT-PCR方法对候选lncRNAs进行初步验证;采用0、0.02、0.05、0.075、0.1和0.2 Gy γ射线照射AHH-1细胞,在照射后4、24、48、72、96和120 h提取总RNA,检测并分析候选lncRNAs的剂量效应关系;取8名健康人的外周血进行0、0.02、0.05、0.075、0.1和0.2 Gy γ射线离体照射,照射后培养24和48 h,进一步验证在人外周血水平上具有辐射响应的lncRNAs表达水平变化情况。结果 转录组测序初步筛选出在0.05和0.1 Gy照射后均发生明显上调或下调的lncRNAs共44个,从中筛选差异表达倍数大于2倍的lncRNAs为SNHG1、SNHG15、NEAT1和PRC1-AS1。在细胞水平上,γ射线照射后4~48 h,与0 Gy相比,PRC1-AS1相对表达水平在0.05、0.075和0.1 Gy 3个剂量点显著升高,差异具有统计学意义(t=-3.11~1.23,P < 0.05);与0 Gy相比,NEAT1相对表达水平在0.02~0.1 Gy剂量范围内明显上调,差异具有统计学意义(t=-2.47~2.10,P < 0.05)。与0 Gy相比,人外周血0~0.2 Gy γ射线照后24 h,PRC1-AS1和NEAT1的相对表达量均明显升高,差异有统计学意义(t=-3.79~-1.96, P < 0.05)。结论 PRC1-AS1和NEAT1表达水平改变有作为低剂量电离辐射生物标志物的潜力。 |
| 英文摘要: |
| Objective To investigate the changes in the expression levels of long non-coding ribonucleic acids (lncRNAs) in human lymphocytes induced by low-dose ionizing radiation (LDIR) and the potential of lncRNAs as radiation biomarkers. Methods Human immortalized lymphocytes (AHH-1) were irradiated with 0, 0.05, and 0.1 Gy of γ-rays at 24 h to extract RNAs for whole transcriptome sequencing. The sequencing was performed based on the 0, 0.05, and 0.1 Gy groups. The differentially expressed lncRNAs induced by LDIR were identified. The molecular functions, biological processes, and signaling pathway enrichment of differentially expressed genes were analyzed through the Gene Ontology (GO) analysis. Candidate lncRNAs were preliminarily validated using the qRT-PCR method. AHH-1 cells were irradiated with 0, 0.02, 0.05, 0.075, 0.1, and 0.2 Gy to extract the total RNAs at 4, 24, 48, 72, 96, and 120 h. The dose-response relationship of candidate lncRNAs was detected and analyzed. Peripheral blood sampled from eight healthy persons was irradiated with 0, 0.02, 0.05, 0.075, 0.1, and 0.2 Gy in vitro, followed by culturing for 24 h and 48 h to further verify the changes in the expression levels of radiation-responsive lncRNAs at the cellular level. Results A total of 44 lncRNAs that were significantly up- or down-regulated after 0.05 and 0.1 Gy irradiation were initially identified through transcriptome sequencing. Among them, lncRNAs with over two-fold differential expression included SNHG1, SNHG15, NEAT1, and PRC1-AS1. At the cellular level, compared to 0 Gy, the relative expression level of PRC1-AS1 after 4 h to 48 h of γ-ray irradiation, was significantly elevated at 0.05, 0.075, and 0.1 Gy(t= -3.11 to 1.23, P < 0.05). In contrast, the relative expression level of NEAT1 was significantly up-regulated in a dose range of 0.02 to 0.1 Gy (t=-2.47 to 2.10, P < 0.05). At the level of human peripheral blood, the relative expression levels of PRC1-AS1 and NEAT1 were significantly increased at 24 h after 0 to 0.2 Gy irradiation (t=-3.79 to -1.96, P < 0.05). Conclusion The PRC1-AS1 and NEAT1 with significant changes in expression levels serve as potential LDIR biomarkers. |
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