USP11在电离辐射诱导肠上皮细胞损伤中的作用初探
Preliminary Study on the Role of USP11 in Radiation-Induced Intestinal Epithelial Cell Injury
王一丹 1焦旸 1曹建平1
作者信息
- 1. 苏州大学医学部放射医学与防护学院，苏州 215123;放射医学与辐射防护国家重点实验室
- 折叠
摘要
目的：初步探究泛素特异性肽酶11（ubiquitin-specific peptidase 11，USP11）在电离辐射诱导肠上皮细胞损伤中的作用及潜在机制。方法：以人肠上皮细胞（HIEC）为研究对象，给予不同剂量及不同时间的电离辐射处理，检测USP11及DNA损伤标志物γ-H2AX的表达变化；构建USP11过表达细胞模型，采用蛋白免疫印迹法（Western blot）和实时荧光定量聚合酶链反应（qRT-PCR）验证模型构建效率；采用免疫荧光和彗星实验评估DNA损伤程度；通过活性氧（reactive oxygen species，ROS）水平检测及线粒体膜电位分析评价氧化应激和线粒体功能变化；进一步检测基础状态下CHK1及其磷酸化CHK1（p-CHK1）的表达变化。结果：电离辐射处理后，USP11在不同剂量和时间条件下均呈总体上调趋势。γ-H2AX水平呈先升高后回落的动态变化，提示细胞在辐射诱导DNA双链断裂后激活损伤应答并启动后续修复过程。USP11过表达显著增强辐照后γ-H2AX荧光信号，增加尾部DNA百分比，促进ROS蓄积，并降低JC-1红/绿荧光比值，提示USP11可加重辐射诱导的DNA损伤、氧化应激及线粒体功能障碍。此外，在未照射状态下，USP11过表达可上调p-CHK1蛋白水平并增加p-CHK1/CHK1比值，而总CHK1蛋白表达无明显变化，提示USP11可能参与基础状态下CHK1磷酸化水平的调控。结论：USP11参与电离辐射诱导的肠上皮细胞损伤过程，其作用可能与促进DNA损伤积累、增强氧化应激、加重线粒体功能障碍及调控CHK1磷酸化相关的损伤应答网络有关。
Abstract
Objective: To preliminarily investigate the role of ubiquitin-specific peptidase 11 (USP11) in ionizing radiation-induced intestinal epithelial cell injury and explore its potential underlying mechanisms.Methods: Human intestinal epithelial cells (HIECs) were exposed to ionizing radiation at different doses and time points, and the expression changes of USP11 and the DNA damage marker γ-H2AX were determined. A USP11-overexpressing cell model was established and verified by Western blot and quantitative real-time polymerase chain reaction (qRT-PCR). DNA damage was evaluated by immunofluorescence staining and comet assay. Oxidative stress and mitochondrial function were assessed by reactive oxygen species (ROS) measurement and mitochondrial membrane potential analysis. In addition, the expression levels of CHK1 and phosphorylated CHK1 (p-CHK1) under basal conditions were examined.Results: Following ionizing radiation exposure, USP11 showed an overall upregulation across different doses and time points.γ-H2AX exhibited a dynamic pattern characterized by an initial increase followed by a subsequent decline, suggesting that cells activated the DNA damage response and initiated the subsequent repair process after radiation-induced DNA double-strand breaks.USP11 overexpression significantly enhanced radiation-induced γ-H2AX fluorescence intensity, increased the percentage of tail DNA, promoted ROS accumulation, and decreased the JC-1 red/green fluorescence ratio, indicating that USP11 aggravated radiation-induced DNA damage, oxidative stress, and mitochondrial dysfunction. Furthermore, under non-irradiated conditions, USP11 overexpression increased p-CHK1 protein levels and the p-CHK1/CHK1 ratio, whereas total CHK1 protein expression remained unchanged, suggesting that USP11 may be involved in the regulation of CHK1 phosphorylation under basal conditions.Conclusion: USP11 is involved in ionizing radiation-induced intestinal epithelial cell injury. Its effects may be associated with enhanced DNA damage accumulation, aggravated oxidative stress, exacerbated mitochondrial dysfunction, and modulation of CHK1 phosphorylation-related damage response networks.关键词
USP11/电离辐射/DNA损伤/氧化应激/线粒体膜电位Key words
USP11/ionizing radiation/DNA damage/oxidative stress/mitochondrial membrane potential引用本文复制引用
王一丹,焦旸,曹建平.USP11在电离辐射诱导肠上皮细胞损伤中的作用初探[EB/OL].(2026-04-01)[2026-04-04].http://www.paper.edu.cn/releasepaper/content/202604-14.学科分类
基础医学/生物科学研究方法、生物科学研究技术
评论