Expanded usage of the Challenge-Comet assay as a DNA repair biomarker in human populations: protocols for fresh and cryopreserved blood samples, and for different challenge agents.

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Abstrakt

Tytuł:: Expanded usage of the Challenge-Comet assay as a DNA repair biomarker in human populations: protocols for fresh and cryopreserved blood samples, and for different challenge agents.
Autorzy:: Valdiglesias V; Centro de Investigaciones Científicas Avanzadas (CICA), Departamento de Biología, Facultad de Ciencias, Universidade da Coruña, Grupo DICOMOSA, Campus A Zapateira s/n, 15071, A Coruña, Spain. .; AE CICA-INIBIC. Oza, Instituto de Investigación Biomédica de A Coruña (INIBIC), 15071, A Coruña, Spain. .
Sánchez-Flores M; AE CICA-INIBIC. Oza, Instituto de Investigación Biomédica de A Coruña (INIBIC), 15071, A Coruña, Spain.; Centro de Investigaciones Científicas Avanzadas (CICA), Departamento de Psicología, Facultad de Ciencias de La Educación, Universidade da Coruña, Grupo DICOMOSA, Campus Elviña s/n, 15071, A Coruña, Spain.
Fernández-Bertólez N; AE CICA-INIBIC. Oza, Instituto de Investigación Biomédica de A Coruña (INIBIC), 15071, A Coruña, Spain.; Centro de Investigaciones Científicas Avanzadas (CICA), Departamento de Psicología, Facultad de Ciencias de La Educación, Universidade da Coruña, Grupo DICOMOSA, Campus Elviña s/n, 15071, A Coruña, Spain.
Au W; University of Medicine, Pharmacy, Science and Technology, Gheorghe Marinescu nr. 38, 540139, Targu Mures, Romania.; University of Texas Medical Branch, Galveston, TX, USA.
Pásaro E; AE CICA-INIBIC. Oza, Instituto de Investigación Biomédica de A Coruña (INIBIC), 15071, A Coruña, Spain.; Centro de Investigaciones Científicas Avanzadas (CICA), Departamento de Psicología, Facultad de Ciencias de La Educación, Universidade da Coruña, Grupo DICOMOSA, Campus Elviña s/n, 15071, A Coruña, Spain.
Laffon B; AE CICA-INIBIC. Oza, Instituto de Investigación Biomédica de A Coruña (INIBIC), 15071, A Coruña, Spain.; Centro de Investigaciones Científicas Avanzadas (CICA), Departamento de Psicología, Facultad de Ciencias de La Educación, Universidade da Coruña, Grupo DICOMOSA, Campus Elviña s/n, 15071, A Coruña, Spain.
Źródło:: Archives of toxicology [Arch Toxicol] 2020 Dec; Vol. 94 (12), pp. 4219-4228. Date of Electronic Publication: 2020 Sep 30.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Original Publication: Berlin, New York, Springer-Verlag.
MeSH Terms:: Biological Monitoring*
Blood Specimen Collection*
Comet Assay*
Cryopreservation*
DNA Damage*
DNA Repair*/drug effects
DNA Repair*/radiation effects
Ultraviolet Rays*
Adult ; Biomarkers/blood ; Bleomycin/toxicity ; Female ; Humans ; Methyl Methanesulfonate/toxicity ; Risk Assessment ; Time Factors ; Young Adult
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Contributed Indexing:: Keywords: Biomonitoring; Cellular repair assay; Challenge-Comet assay; DNA repair; Genotoxicity; Health hazard assessment
Substance Nomenclature:: 0 (Biomarkers)
11056-06-7 (Bleomycin)
AT5C31J09G (Methyl Methanesulfonate)
Entry Date(s):: Date Created: 20201001 Date Completed: 20210803 Latest Revision: 20210803
Update Code:: 20240105
DOI:: 10.1007/s00204-020-02881-5
PMID:: 33000292
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Deficiencies in DNA damage response and repair (DDRR) can cause serious pathological outcomes; therefore, having an ability to determine individual DDRR would enhance specificities in health risk assessment and in determining individual's response to cancer therapies. However, most methods for evaluating DDRR are not fully appropriate for population studies. The Challenge-Comet assay has gained acceptance for this purpose. The assay has traditionally used X-rays as challenge agent and isolated peripheral blood mononuclear cells (PBMC) as cell specimen. To enhance the usefulness of the assay, the objectives of this investigation were to use differently processed blood samples, to employ other challenge agents with different mechanisms of induction of DNA damage/repair, and to generate protocols for detecting different DDRR capacities. Fresh and frozen blood samples were challenged with bleomycin, methyl methanesulfonate (MMS) and ultraviolet light. Significant induction of damage after all treatments, and progressive and time-dependent DDRR were observed. No significant differences were obtained in the DDRR capacities of fresh or frozen whole blood samples as compared to PBMC, except that fresh blood samples showed higher MMS-induced DDRR capacity than PBMC. Results from this study show that the Challenge-Comet assay can be used as routine biomarker of DDRR capacity in human biomonitoring studies, and that whole blood is also a useful biomatrix for this assay. The collected data allow us to recommend different protocols for the Challenge-Comet assay which are useful for evaluating DDRR capacities in several key DNA repair pathways. Consequently, the usefulness of the Challenge-Comet assay can be greatly expanded.

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