Evaluation of DNA double-strand break repair capacity in human cells: Critical overview of current functional methods

Tatin, Xavier; Muggiolu, Giovanna; Sauvaigo, Sylvie; Breton, Jean

doi:10.1016/j.mrrev.2021.108388

Cited by 13 publications

(6 citation statements)

References 340 publications

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“…Identifying the targets of a chemical using in vitro analysis is complicated, costly, and time-consuming and does not necessarily reflect its physiological effects in cells in vivo. Comet assays, pulse-field gel electrophoresis, and single-molecule Förster resonance energy transfer are often used to elucidate the DNA strand-break-inducing effects of any chemical [12][13][14]. However, it is difficult to establish the required experimental conditions.…”

Section: Introductionmentioning

confidence: 99%

A Cell System-Assisted Strategy for Evaluating the Natural Antioxidant-Induced Double-Stranded DNA Break (DSB) Style

Someya

Kobayashi

Toriumi

et al. 2023

Genes

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Natural antioxidants derived from plants exert various physiological effects, including antitumor effects. However, the molecular mechanisms of each natural antioxidant have not yet been fully elucidated. Identifying the targets of natural antioxidants with antitumor properties in vitro is costly and time-consuming, and the results thus obtained may not reliably reflect in vivo conditions. Therefore, to enhance understanding regarding the antitumor effects of natural antioxidants, we focused on DNA, one of the targets of anticancer drugs, and evaluated whether antioxidants, e.g., sulforaphane, resveratrol, quercetin, kaempferol, and genistein, which exert antitumor effects, induce DNA damage using gene-knockout cell lines derived from human Nalm-6 and HeLa cells pretreated with the DNA-dependent protein kinase inhibitor NU7026. Our results suggested that sulforaphane induces single-strand breaks or DNA strand crosslinks and that quercetin induces double-strand breaks. In contrast, resveratrol showed the ability to exert cytotoxic effects other than DNA damage. Our results also suggested that kaempferol and genistein induce DNA damage via unknown mechanisms. Taken together, the use of this evaluation system facilitates the analysis of the cytotoxic mechanisms of natural antioxidants.

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Section: Introductionmentioning

confidence: 99%

A Cell System-Assisted Strategy for Evaluating the Natural Antioxidant-Induced Double-Stranded DNA Break (DSB) Style

Someya

Kobayashi

Toriumi

et al. 2023

Genes

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“…Platinum‐DNA adduct accumulation is a determinant step for the cytotoxicity of platinum‐based antitumor agents which lead to the destabilization of double helix, blocking replication and inhibiting transcription 6 . However, DNA repair capacity, which varies widely among individuals, plays a fundamental role in timely removing DNA adducts 7 . In general, a high DNA repair capacity of tumour cells is a warning of potential chemotherapy resistance of NSCLC patients 8 .…”

Section: Introductionmentioning

confidence: 99%

“…6 However, DNA repair capacity, which varies widely among individuals, plays a fundamental role in timely removing DNA adducts. 7 In general, a high DNA repair capacity of tumour cells is a warning of potential chemotherapy resistance of NSCLC patients. 8 As known, there are at least several DNA repair systems in the human body, such as nucleotide excision repair (NER), base excision repair (BER) and mismatch repair (MMR).…”

Section: Introductionmentioning

confidence: 99%

A miR‐15a related polymorphism affects NSCLC prognosis via altering ERCC1 repair to platinum‐based chemotherapy

Xue

Zhang

et al. 2022

J Cellular Molecular Medi

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Platinum-based chemotherapy is regarded as a preferential curative-intent option for non-small cell lung cancer (NSCLC), while the acquired drug resistance has become a major obstacle that limits its clinical application. Since the repair efficiency of tumour cells to platinum-DNA adducts plays a crucial role in chemotherapy resistance, we aimed to explore whether several meaningful polymorphisms of DNA repair genes were associated with the benefits of platinum-based chemotherapy in NSCLC patients. Firstly, six single nucleotide polymorphisms (SNPs) located in the 3'untranslated region (3'UTR) of three DNA repair genes were detected in 246 NSCLC patients receiving platinum-based chemotherapy and analysed the correlation of these candidate SNPs with the overall survival. Cox proportional hazard model showed that NSCLC patients carrying ERCC1 rs3212986 AA genotype had a shorter overall survival compared to those with CC. Mechanistically, we performed tumour chemosensitivity assay to observe the convincing linkage of rs3212986 polymorphism with ERCC1 expression and cisplatin sensitivity. The subsequent in vitro experiments identified that rs3212986 polymorphism altered the post-transcriptional regulation of ERCC1 via affecting the binding of miR-15a, and further changed the sensitivity to platinum analogue. It reminded that patients carrying ERCC1 rs3212986 CC homozygote were expected to respond better to platinum-based chemotherapy due to a lower expression of ERCC1. Compared with previous studies, our current comprehensive study suggested that rs3212986, a 3'UTR polymorphism in ERCC1, might have clinical relevance in predicting the prognosis of NSCLC patients receiving platinum-based chemotherapy.

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“…Therefore, systematic diagnosis of DDR defects should be part of the ongoing strategy of care personalization to better identify the most optimal treatment modality for a particular individual. DDR capacities can be estimated by multiple methods, more or less sophisticated, at the gene, transcript, or protein levels 19 , 20 . However, no existing method measuring the functionality of DSBs repair meets all the criteria required for practical adoption in routine, neither for research nor for clinical applications.…”

Section: Introductionmentioning

confidence: 99%

A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair

Tatin

Muggiolu²,

Libert³

et al. 2022

Sci Rep

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The repair of DNA double-strand breaks (DSBs) involves interdependent molecular pathways, of which the choice is crucial for a cell’s fate when facing a damage. Growing evidence points toward the fact that DSB repair capacities correlate with disease aggressiveness, treatment response and treatment-related toxicities in cancer. Scientific and medical communities need more easy-to-use and efficient tools to rapidly estimate DSB repair capacities from a tissue, enable routine-accessible treatment personalization, and hopefully, improve survival. Here, we propose a new functional biochip assay (NEXT-SPOT) that characterizes DSB repair-engaged cellular pathways and provides qualitative and quantitative information on the contribution of several pathways in less than 2 h, from 10 mg of cell lysates. We introduce the NEXT-SPOT technology, detail the molecular characterizations of different repair steps occurring on the biochip, and show examples of DSB repair profiling using three cancer cell lines treated or not with a DSB-inducer (doxorubicin) and/or a DNA repair inhibitor (RAD51 inhibitor; DNA-PK inhibitor; PARP inhibitor). Among others, we demonstrate that NEXT-SPOT can accurately detect decreased activities in strand invasion and end-joining mechanisms following DNA-PK or RAD51 inhibition in DNA-PK-proficient cell lines. This approach offers an all-in-one reliable strategy to consider DSB repair capacities as predictive biomarkers easily translatable to the clinic.

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Evaluation of DNA double-strand break repair capacity in human cells: Critical overview of current functional methods

Cited by 13 publications

References 340 publications

A Cell System-Assisted Strategy for Evaluating the Natural Antioxidant-Induced Double-Stranded DNA Break (DSB) Style

A Cell System-Assisted Strategy for Evaluating the Natural Antioxidant-Induced Double-Stranded DNA Break (DSB) Style

A miR‐15a related polymorphism affects NSCLC prognosis via altering ERCC1 repair to platinum‐based chemotherapy

A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair

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