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An Analytical Method for Quantifying the Yields of DNA Double-Strand Breaks Coupled with Strand Breaks by gamma-H2AX Focus Formation Assay Based on Track-Structure Simulation

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Title: An Analytical Method for Quantifying the Yields of DNA Double-Strand Breaks Coupled with Strand Breaks by gamma-H2AX Focus Formation Assay Based on Track-Structure Simulation
Authors: Yachi, Yoshie Browse this author
Matsuya, Yusuke Browse this author →KAKEN DB
Yoshii, Yuji Browse this author
Fukunaga, Hisanori Browse this author →KAKEN DB
Date, Hiroyuki Browse this author →KAKEN DB
Kai, Takeshi Browse this author →KAKEN DB
Keywords: Monte Carlo track-structure simulation
complex DSB
gamma-H2AX focus formation assay
photon irradiation
Issue Date: 10-Jan-2023
Publisher: MDPI
Journal Title: International Journal of Molecular Sciences
Volume: 24
Issue: 2
Start Page: 1386
Publisher DOI: 10.3390/ijms24021386
PMID: 36674901
Abstract: Complex DNA double-strand break (DSB), which is defined as a DSB coupled with additional strand breaks within 10 bp in this study, induced after ionizing radiation or X-rays, is recognized as fatal damage which can induce cell death with a certain probability. In general, a DSB site inside the nucleus of live cells can be experimentally detected using the gamma-H2AX focus formation assay. DSB complexity is believed to be detected by analyzing the focus size using such an assay. However, the relationship between focus size and DSB complexity remains uncertain. In this study, using Monte Carlo (MC) track-structure simulation codes, i.e., an in-house WLTrack code and a Particle and Heavy Ion Transport code System (PHITS), we developed an analytical method for qualifying the DSB complexity induced by photon irradiation from the microscopic image of gamma-H2AX foci. First, assuming that events (i.e., ionization and excitation) potentially induce DNA strand breaks, we scored the number of events in a water cube (5.03 x 5.03 x 5.03 nm(3)) along electron tracks. Second, we obtained the relationship between the number of events and the foci size experimentally measured by the gamma-H2AX focus formation assay. Third, using this relationship, we evaluated the degree of DSB complexity induced after photon irradiation for various X-ray spectra using the foci size, and the experimental DSB complexity was compared to the results estimated by the well-verified DNA damage estimation model in the PHITS code. The number of events in a water cube was found to be proportional to foci size, suggesting that the number of events intrinsically related to DSB complexity at the DNA scale. The developed method was applicable to focus data measured for various X-ray spectral situations (i.e., diagnostic kV X-rays and therapeutic MV X-rays). This method would contribute to a precise understanding of the early biological impacts of photon irradiation by means of the gamma-H2AX focus formation assay.
Type: article
URI: http://hdl.handle.net/2115/88582
Appears in Collections:保健科学院・保健科学研究院 (Graduate School of Health Sciences / Faculty of Health Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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