Comet Assay analysis of DNA strand breaks after exposure to the DNA-incorporated Auger Electron Emitter Iodine-125.

IF 2.1 4区 医学 Q2 BIOLOGY International Journal of Radiation Biology Pub Date : 2023-01-01 DOI:10.1080/09553002.2020.1851059
Marcus Unverricht-Yeboah, Kathrin Holtmann, Ralf Kriehuber
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引用次数: 4

Abstract

Purpose: Ionizing radiation causes various types of DNA damage e.g. single strand breaks (SSB) and double strand breaks (DSB), whereby the SSB/DSB ratio is shifted toward the DSB with increasing LET. For the DNA-incorporated Auger electron emitter Iodine-125 a SSB/DSB ratio of 5.4:1 is calculated based on computer simulations. In the presented work the SSB/DSB ratio of DNA-incorporated Iodine-125 was experimentally determined and compared to external homogenous γ-irradiation.

Materials and methods: Iodine-125-iododeoxyuridine (I-125-UdR) was incorporated into the DNA of SCL-II cells and cells were subsequently frozen for decay accumulation. Accordingly, external γ-irradiation (Cs-137) experiments were performed in frozen cells. After exposure the neutral or alkaline Comet Assay was performed to quantify DSB or DSB and SSB, respectively. Automated quantification of the comets was performed using the Olive Tail Moment (Metafer CometScan; MetaSystems). Calculation of absorbed dose for Auger electrons on cellular level is extremely biased due to the exclusive DNA localization of I-125-UdR. To avoid dose calculation the γ-H2AX assay was used in order to allow the comparison of the Comet Assay data between both investigated radiation qualities.

Results: For low-LET γ-radiation, a SSB/DSB ratio of 10:1 was determined. In contrast, a lower SSB/DSB ratio of 6:1 was induced by DNA-incorporated Iodine-125 which compares very well to the calculated values of Pomplun and co-authors.

Conclusion: DNA-incorporated Iodine-125 induces a high-LET type DNA damage pattern in respect to SSB/DSB ratio.

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暴露于含DNA的俄歇电子发射器碘-125后DNA链断裂的彗星分析。
目的:电离辐射引起多种类型的DNA损伤,如单链断裂(SSB)和双链断裂(DSB),其中SSB/DSB比值随着LET的增加而向DSB偏移。在计算机模拟的基础上,计算出含dna的俄歇电子发射器碘- 125a的SSB/DSB比为5.4:1。本文通过实验测定了dna结合碘-125的SSB/DSB比值,并与外均质γ辐照进行了比较。材料和方法:将碘-125-碘脱氧尿苷(I-125-UdR)掺入SCL-II细胞的DNA中,冷冻细胞进行衰变积累。因此,在冷冻细胞中进行体外γ-辐照(Cs-137)实验。暴露后进行中性或碱性彗星试验分别定量DSB或DSB和SSB。使用Olive Tail Moment (Metafer CometScan;MetaSystems)。由于I-125-UdR的DNA定位,在细胞水平上计算俄歇电子的吸收剂量存在极大的偏差。为了避免剂量计算,使用了γ-H2AX测定法,以便在两种研究的辐射质量之间进行彗星测定数据的比较。结果:对于低let γ辐射,SSB/DSB比值为10:1。相比之下,dna结合碘-125诱导的SSB/DSB比较低,为6:1,与Pomplun及其合作者的计算值相当。结论:DNA掺入的碘-125诱导高let型DNA损伤模式,与SSB/DSB比值有关。
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来源期刊
CiteScore
5.00
自引率
11.50%
发文量
142
审稿时长
3 months
期刊介绍: The International Journal of Radiation Biology publishes original papers, reviews, current topic articles, technical notes/reports, and meeting reports on the effects of ionizing, UV and visible radiation, accelerated particles, electromagnetic fields, ultrasound, heat and related modalities. The focus is on the biological effects of such radiations: from radiation chemistry to the spectrum of responses of living organisms and underlying mechanisms, including genetic abnormalities, repair phenomena, cell death, dose modifying agents and tissue responses. Application of basic studies to medical uses of radiation extends the coverage to practical problems such as physical and chemical adjuvants which improve the effectiveness of radiation in cancer therapy. Assessment of the hazards of low doses of radiation is also considered.
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