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彗星芯片分析人原代淋巴细胞可定量评估氧化 DNA 损伤修复动力学的个体间差异。

CometChip analysis of human primary lymphocytes enables quantification of inter-individual differences in the kinetics of repair of oxidative DNA damage.

机构信息

Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA; MIT Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.

Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA; MIT Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA; Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, MA, 02115, USA.

出版信息

Free Radic Biol Med. 2021 Oct;174:89-99. doi: 10.1016/j.freeradbiomed.2021.07.033. Epub 2021 Jul 26.

Abstract

Although DNA repair is known to impact susceptibility to cancer and other diseases, relatively few population studies have been performed to evaluate DNA repair kinetics in people due to the difficulty of assessing DNA repair in a high-throughput manner. Here we use the CometChip, a high-throughput comet assay, to explore inter-individual variation in repair of oxidative damage to DNA, a known risk factor for aging, cancer and other diseases. DNA repair capacity after HO-induced DNA oxidation damage was quantified in peripheral blood mononuclear cells (PBMCs). For 10 individuals, blood was drawn at several times over the course of 4-6 weeks. In addition, blood was drawn once from each of 56 individuals. DNA damage levels were quantified prior to exposure to HO and at 0, 15, 30, 60, and 120-min post exposure. We found that there is significant variability in DNA repair efficiency among individuals. When subdivided into quartiles by DNA repair efficiency, we found that the average t is 81 min for the slowest group and 24 min for the fastest group. This work shows that the CometChip can be used to uncover significant differences in repair kinetics among people, pointing to its utility in future epidemiological and clinical studies.

摘要

虽然已知 DNA 修复会影响癌症和其他疾病的易感性,但由于难以以高通量方式评估 DNA 修复,因此相对较少进行评估人类 DNA 修复动力学的人群研究。在这里,我们使用 CometChip(一种高通量彗星测定法)来探索 DNA 氧化损伤修复的个体间差异,这是衰老、癌症和其他疾病的已知风险因素。外周血单核细胞 (PBMC) 中的 HO 诱导的 DNA 氧化损伤后的 DNA 修复能力被定量。对于 10 个人,在 4-6 周的过程中多次抽取血液。此外,还从 56 个人中每人抽取一次血液。在暴露于 HO 之前和暴露后 0、15、30、60 和 120 分钟时,定量了 DNA 损伤水平。我们发现个体之间的 DNA 修复效率存在显着差异。当按 DNA 修复效率细分为四分之一时,我们发现最慢组的平均 t 为 81 分钟,最快组的平均 t 为 24 分钟。这项工作表明 CometChip 可用于揭示人群中修复动力学的显着差异,表明其在未来的流行病学和临床研究中的实用性。

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