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

Yachi, Yoshie; Matsuya, Yusuke; Yoshii, Yukie; Fukunaga, Hisanori; Date, Hiroyuki; Kai, Takeshi

doi:10.3390/ijms24021386

Cited by 4 publications

(4 citation statements)

References 40 publications

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“…In the present study, the increased accumulation of γ-H2AX of irradiated OCYs suggested that IR caused double-strand breaks in DNA. Prior studies have demonstrated that temporally unrepaired DSBs can lead to termination segregation, and the end misconnection of these broken DNAs usually gives rise to chromosomal aberrations [31]. This study further revealed a marked increase in the characteristic heterochromatin structures in the nuclei of irradiated OCYs as punctate aggregates.…”

Section: Discussionsupporting

confidence: 60%

“…After exposure to IR, there are large clusters of DSBs in the cell, followed by a series of stress responses [30,31]. DSBs promote the recruitment and binding of kinases from ATM to sites of DNA damage, thereby driving the phosphorylation of histone H2AX to form γ-H2AX, which in turn facilitates the assembly of specific DNA repair complexes [32].…”

Section: Discussionmentioning

confidence: 99%

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C-C Motif Chemokine Ligand 5 (CCL5) Promotes Irradiation-Evoked Osteoclastogenesis

Wang,

Zhao,

et al. 2023

IJMS

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The imbalance that occurs in bone remodeling induced by irradiation (IR) is the disruption of the balance between bone formation and bone resorption. In this study, primary osteocytes (OCYs) of femoral and tibial origin were cultured and irradiated. It was observed that irradiated OCY showed extensive DNA damage, which led to the initiation of a typical phenotype of cellular senescence, including the secretion of senescence-associated secretory phenotype (SASP), especially the C-C motif chemokine ligand 5 (CCL5). In order to explore the regulation of osteoclastogenic potential by IR-induced senescent OCYs exocytosis factor CCL5, the conditioned medium (CM) of OCYs was co-cultured with RAW264.7 precursor cells. It was observed that in the irradiated OCY co-cultured group, the migration potential increased compared with the vehicle culture group, accompanied by an enhancement of typical mature OCs; the expression of the specific function of enzyme tartrate-resistant acid phosphatase (TRAP) increased; and the bone-destructive function was enhanced. However, a neutralizing antibody to CCL5 could reverse the extra-activation of osteoclastogenesis. Accordingly, the overexpression of p-STAT3 in irradiated OCY was accompanied by CCL5. It was concluded that CCL5 is a potential key molecule and the interventions targeting CCL5 could be a potential strategy for inhibiting osteoclastogenesis and restoring bone remodeling.

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Section: Discussionsupporting

confidence: 60%

Section: Discussionmentioning

confidence: 99%

C-C Motif Chemokine Ligand 5 (CCL5) Promotes Irradiation-Evoked Osteoclastogenesis

Wang,

Zhao,

et al. 2023

IJMS

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“…The DNA damage response system, which involves γ-H2AX, is crucial in preserving genomic integrity by signaling and facilitating the repair of DNA double-strand breaks ( 25 , 26 ). When the DNA of a cell undergoes DSB, the signaling and detection of the damaged site stimulate the formation of γ-H2AX, which exerts a vital effect in response to DNA damage and also involves DNA repair ( 27 ). The results of this study showed that with the effect of ETO, the expression of γ-H2AX and ARID1A was significantly increased.…”

Section: Discussionmentioning

confidence: 99%

ARID1A is involved in DNA double-strand break repair in gastric cancer

Zhang,

Qian,

et al. 2024

J Gastrointest Oncol

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Background Defects in DNA damage repair can cause genetic mutations, which in turn can cause different types of cancers. Chromatin remodeling complexes, which help repair damaged DNA, can cause the chromatin structure to change as a result of DNA damage. ARID1A may play a role in the process of DNA damage repair, and arid1a may be related to the occurrence and development of gastric cancer (GC). This study aimed to investigate the mechanism of ARID1A regulating the DNA damage repair of gastric adenocarcinoma cell lines AGS and SGC-7901 and its effect on migration, proliferation and apoptosis. Methods The expression of ARID1A plasmid was detected by Western blot and real-time polymerase chain reaction (PCR). The effect of etoposide (ETO) on the survival rate of AGS and SGC-7901 gastric adenocarcinoma cell lines was detected by MTT assay. The DNA double-strand break model was established by ETO and then passed through the comet assay and immunofluorescence co-localization to observe DNA damage; western blot method was used to detect the effect of ARID1A on the expression of related proteins in DNA damage repair pathway in gastric adenocarcinoma cells; scratch test and colony formation experiments were used to observe ARID1A migration and proliferation of gastric adenocarcinoma cells. The flow cytometry was used to detect the effect of ARID1A on apoptosis of gastric adenocarcinoma cells. Results The expression of mRNA and protein was increased after transfection of ARID1A plasmid. ETO was confirmed by MTT assay to inhibit cell survival in a dose-dependent manner. After the DNA double-strand break model was established by ETO, the expression levels of phospho-ataxia telangiectasia mutated (p-ATM) protein increased in the overexpressed ARID1A group. Meanwhile, the overexpressed ARID1A group had a shortened tail moment, and γ-H2AX and ARID1A co-localized in the DNA damage site of the nucleus. The over-expressed ARID1A group had weaker wound healing ability, reduced number of clone formation, and increased apoptosis rate. Conclusions ARID1A may repair DNA double-strand breaks caused by ETO by p-ATM pathway; ARID1A can inhibit the migration and proliferation of gastric adenocarcinoma cells and promote apoptosis. Our findings indicate that ARID1A could serve as a therapeutic target and biomarker for GC patients.

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“…Crucially, increased cancer risks have been extensively reported [2,7,8]. Ionizing radiation exposure leads to chromosomal aberrations [9], DNA damage [10,11], alterations in the cell cycle [12,13], and apoptosis [14,15]. However, the complex nature and small astronaut cohort have made the research on space radiation protection challenging and the results highly uncertain [16].…”

Section: Introductionmentioning

confidence: 99%

Drug Discovery Targeting Post-Translational Modifications in Response to DNA Damages Induced by Space Radiation

Xie

Zhou

2023

IJMS

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DNA damage in astronauts induced by cosmic radiation poses a major barrier to human space exploration. Cellular responses and repair of the most lethal DNA double-strand breaks (DSBs) are crucial for genomic integrity and cell survival. Post-translational modifications (PTMs), including phosphorylation, ubiquitylation, and SUMOylation, are among the regulatory factors modulating a delicate balance and choice between predominant DSB repair pathways, such as non-homologous end joining (NHEJ) and homologous recombination (HR). In this review, we focused on the engagement of proteins in the DNA damage response (DDR) modulated by phosphorylation and ubiquitylation, including ATM, DNA-PKcs, CtIP, MDM2, and ubiquitin ligases. The involvement and function of acetylation, methylation, PARylation, and their essential proteins were also investigated, providing a repository of candidate targets for DDR regulators. However, there is a lack of radioprotectors in spite of their consideration in the discovery of radiosensitizers. We proposed new perspectives for the research and development of future agents against space radiation by the systematic integration and utilization of evolutionary strategies, including multi-omics analyses, rational computing methods, drug repositioning, and combinations of drugs and targets, which may facilitate the use of radioprotectors in practical applications in human space exploration to combat fatal radiation hazards.

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

Cited by 4 publications

References 40 publications

C-C Motif Chemokine Ligand 5 (CCL5) Promotes Irradiation-Evoked Osteoclastogenesis

C-C Motif Chemokine Ligand 5 (CCL5) Promotes Irradiation-Evoked Osteoclastogenesis

ARID1A is involved in DNA double-strand break repair in gastric cancer

Drug Discovery Targeting Post-Translational Modifications in Response to DNA Damages Induced by Space Radiation

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