In vivo Comet assay on isolated kidney cells to distinguish genotoxic carcinogens from epigenetic carcinogens or cytotoxic compounds

Nesslany, Fabrice; Zennouche, Nadia; Simar-Meintières, Sophie; Talahari, Ismaïl; NKili-Mboui, Esther-Nadège; Marzin, Daniel

doi:10.1016/j.mrgentox.2007.02.010

Cited by 68 publications

(29 citation statements)

References 70 publications

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“…Crosslinks can be detected by increasing the duration of electrophoresis to such an extend that control cells exhibit significant DNA migration, demonstrating the effect of crosslinking by a retardation in the extent of DNA migration in comparison with the control. Therefore, in order to ensure that DON do not induced crosslinks, it was decided to increase the duration of electrophoresis in the next study (Nesslany et al, 2007) (see Section 9.2).…”

Section: Hogg1-modified Comet Assay On Blood Cells Of Mice Exposed Tomentioning

confidence: 99%

The in vivo genotoxicity studies on nivalenol and deoxynivalenol

Hégarat

Takakura

Simar

et al. 2014

EFSA Supporting Publications

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Nivalenol (NIV) and deoxynivalenol (DON) are structurally related mycotoxins produced by Fusarium fungi. These fungi typically infest cereal crops such as wheat, maize, barley, oats and rye, and NIV and DON are regularly found in cereal grains, food and feed. Recent risk assessments identified possible data gaps for both DON and NIV in particular with respect to genotoxicity and carcinogenicity. The overall objective of the project was to assess the genotoxicity of DON and NIV, including the identification of potential modes of action. A battery of in vivo genotoxicity tests was performed in mice: Comet assay with and without fpg in seven organs (duodenum, colon, blood, liver, spleen, kidney, bone marrow), micronucleus assay in bone marrow and colon, and Pig-a assay in peripheral blood. In addition, to clarify the genotoxic mode of action of both mycotoxins, we performed in vitro Comet assay studies in TK6 cells to investigate potential genotoxic oxidative stress induced by mycotoxins. The response in all the genotoxicity assays with NIV after three oral doses at 5, 10 and 20 mg/kg, were uniformly negative. In the case of DON, we found that DON failed to induce micronuclei formation in bone marrow and colon and failed to induce DNA damage in all organs observed by the Comet assay with and without fpg at 4, 8 and 16 mg/kg. The Pig-a assay with DON after three oral gavage doses at 2, 4 and 8 mg/kg, did not show any mutagenic effect at day 28 and 45 after the last dose. In vitro studies indicated that both mycotoxins did not induce DNA damage in the Comet assay with or without fpg in TK6 cells even after GSH depletion. It was concluded that NIV and DON could be considered as devoid of genotoxic potential and pose no genotoxic or mutagenic risk.

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Section: Hogg1-modified Comet Assay On Blood Cells Of Mice Exposed Tomentioning

confidence: 99%

The in vivo genotoxicity studies on nivalenol and deoxynivalenol

Hégarat

Takakura

Simar

et al. 2014

EFSA Supporting Publications

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“…This test was performed following Nesslany et al [2007] by the oral route using a single treatment followed by 2 sampling times of 3-6 or 22-26 hr.…”

Section: In Vivo Rodent Comet Assay In Rat Kidneymentioning

confidence: 99%

“…At each recommended expression time [Tice et al, 2000], treated animals were sacrificed and the cells of the target organ were isolated as previously described [Nesslany et al, 2007]. The viability of the resulting cell suspensions was determined using trypan blue technique (5 vol.…”

Section: Treatmentmentioning

confidence: 99%

“…For each cell suspension, at least 3 slides were prepared as previously described [Nesslany et al, 2007]. Just prior to score, the DNA was stained using Propidium Iodide (20 lg/ml in distilled water; 25 ll/slide).…”

Section: Slide Preparation and Countingmentioning

confidence: 99%

“…Therefore, it was deemed necessary to assess the in vivo primary DNA damage potential of NTA in the rodent Comet assay on isolated kidney cells able to discriminate genotoxic from epigenetic carcinogens and to avoid the interference of cytotoxicity [Nesslany et al, 2007]. Next, three in vitro tests using the maximum concentration compatible with the solubility and/or the cytotoxicity of NTA were implemented: the Ames test using TA1537, TA98, TA100 and TA102 tester strains, the in vitro micronucleus assay on L5178Y mouse lymphoma cells and on CTLL2/CTLL2 Bcl2 cells performed concurrently to the apoptosis assessment.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the genotoxicity of nitrilotriacetic acid

Nesslany

Simar-Meintières

Watzinger

et al. 2008

Environ and Mol Mutagen

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The chelating agent nitrilotriacetic acid (NTA), classified as an epigenetic rodent carcinogen, was assessed in the in vivo rodent Comet assay on isolated kidney cells. Unexpected potent increases in DNA damage were obtained in both the short (3-6 hr) and long-term (22-26 hr) expression times after a single oral treatment at 1,000-2,000 mg/kg bw. NTA was assayed in the Ames test using TA1537, TA98, TA100, and TA102 tester strains, and in the in vitro micronucleus assay on L5178Y mouse lymphoma cells and on CTLL2 and CTLL2/Bcl2 cells coupled to the apoptosis measurement, both with and without metabolic activation by aroclor 1254-induced liver or kidney rat S9-mix. Whatever the S9 origin, neither genotoxicity nor apoptosis was detected, while a strong increase in the micronuclei formation was observed without S9 without any apoptosis induction. The direct genotoxicity of NTA was confirmed in the mouse lymphoma tk+/- gene mutation assay and in the chromosomal aberrations test on human lymphocytes. When tested in combination with an excess of Ca2+, NTA gave negative results on L5178Y mouse lymphoma cells in the in vitro Comet and micronucleus assays but still induced DNA damage on rat primary kidney cells. The higher sensitivity of renal cells to Ca2+ variations could explained the positive response observed in vivo. The carcinogenicity of NTA could be a consequence of the survival of kidney cells to intracellular variations of Ca2+, leading to a local and indirect genotoxicity. This suggests that threshold dose exists beyond which tumor-generating events will be displayed.

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D‐Limonen [MAK Value Documentation in German language, 2012]

2012

The MAK‐Collection for Occupational Health and Safety

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Veröffentlicht in der Reihe Gesundheitsschädliche Arbeitsstoffe , 52. Lieferung, Ausgabe 2012 Der Artikel enthält folgende Kapitel: Wirkungsmechanismus Toxikokinetik und Metabolismus Entwicklungstoxizität Genotoxizität Bewertung

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In vivo Comet assay on isolated kidney cells to distinguish genotoxic carcinogens from epigenetic carcinogens or cytotoxic compounds

Cited by 68 publications

References 70 publications

The in vivo genotoxicity studies on nivalenol and deoxynivalenol

The in vivo genotoxicity studies on nivalenol and deoxynivalenol

Characterization of the genotoxicity of nitrilotriacetic acid

D‐Limonen [MAK Value Documentation in German language, 2012]

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