Single cell gel electrophoresis (SCGE) and Pig-a mutation assay in vivo-tools for genotoxicity testing from a regulatory perspective: a study of benzo[a]pyrene in Ogg1(-/-) mice.
Single cell gel electrophoresis (SCGE) and Pig-a mutation assay in vivo-tools for genotoxicity testing from a regulatory perspective: a study of benzo[a]pyrene in Ogg1(-/-) mice.
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DOI：
10.1016/j.mrgentox.2014.07.010
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发表时间：
2014-09-15
影响因子：
1.9
通讯作者：
Olsen, Ann-Karin
中科院分区：
文献类型：
--
作者：
Graupner, Anne;Instanes, Christine;Dertinger, Stephen D.;Andersen, Jill Mari;Lindeman, Birgitte;Rongved, Tonje Danielsen;Brunborg, Gunnar;Olsen, Ann-Karin
关键词：
经合组织制定了测试指南（TG），以确定具有遗传毒性作用的药剂。体内碱性单细胞凝胶电泳（SCGE）试验目前正在准备成为这样的TG。在一项SCGE/Pig-a基因突变联合研究中，研究人员使用已知可诱导生殖细胞突变的典型基因毒物苯并[a]芘（BaP）暴露水平进行了评估。我们的目的是更好地了解(i)两种方法在血液中的优缺点及其预测生殖细胞诱变性的潜力，以及（ii）体内bap暴露后活性氧（ROS）的参与。为了探索活性氧与BaP遗传毒性的关系，我们使用了DNA糖基化酶缺失的小鼠模型。具体来说，C57BL/6小鼠（Ogg1+/+和Ogg1−/−）连续3天服用50 mg BaP/kg/天。在最后一次处理后4小时，用甲脒嘧啶DNA糖基酶（Fpg）和不加Fpg的SCGE测定有核血细胞的DNA损伤。在治疗后2周和4周对猪a突变型红细胞进行分析。在两种版本的SCGE检测中，bap诱导的DNA病变均未显着增加。两周后，未成熟红细胞和成熟红细胞的表型突变频率显著增加。这些效应不受基因型的影响，这表明氧化损伤可能在BaP遗传毒性中起次要作用，至少在本研究的急性暴露情况下是这样。虽然这两种检测方法都是很有前途的风险评估工具，但这些结果强调了了解每种检测方法检测化学物质遗传毒性潜力的局限性的必要性。
The OECD has developed test guidelines (TG) to identify agents with genotoxic effects. The in vivo alkaline single cell gel electrophoresis (SCGE) assay is currently being prepared to become such a TG. The performance of a combined SCGE/Pig-a gene mutation study was evaluated with the prototypical genotoxicant benzo[a]pyrene (BaP) at an exposure level known to induce germ cell mutation. We aimed to better understand (i) the strengths and weaknesses of the two methods applied in blood and their potential to predict germ cell mutagenicity, and (ii) the involvement of reactive oxygen species (ROS) following in vivo BaP-exposure. To explore the involvement of ROS on BaP genotoxicity, we utilized a mouse model deficient in a DNA glycosylase. Specifically, C57BL/6 mice (Ogg1+/+ and Ogg1−/−) were treated for three consecutive days with 50 mg BaP/kg/day. DNA damage in nucleated blood cells was measured four hours after the last treatment with the SCGE assay, with and without Formamidopyrimidine DNA glycosylase (Fpg). Pig-a mutant phenotype blood erythrocytes were analysed two and four weeks after treatment. BaP-induced DNA lesions were not significantly increased in either version of the SCGE assay. The phenotypic mutation frequencies for immature and mature erythrocytes were significantly increased after two weeks. These effects were not affected by genotype, suggesting oxidative damage may have a minor role in BaP genotoxicity, at least in the acute exposure situation studied here. While both assays are promising tools for risk assessment, these results highlight the necessity of understanding the limitations regarding each assay’s ability to detect chemicals’ genotoxic potential.
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影响因子：
3.7
作者：
Olsen AK;Andreassen A;Singh R;Wiger R;Duale N;Farmer PB;Brunborg G
通讯作者：
Brunborg G
DOI：
10.1016/j.mrgentox.2008.11.006
发表时间：
2009-02-19
影响因子：
1.9
作者：
Dobrovolsky, Vasily N.;Shaddock, Joseph G.;Morris, Suzanne M.
通讯作者：
Morris, Suzanne M.
DOI：
10.1016/j.mrgentox.2011.02.009
发表时间：
2011-05-18
影响因子：
1.9
作者：
Bowen, Damian E.;Whitwell, James H.;Kirkland, David J.
通讯作者：
Kirkland, David J.
DOI：
10.1016/j.mrgentox.2009.12.012
发表时间：
2010-02-02
影响因子：
1.9
作者：
Hansen, Siri Helland;Olsen, Ann Karin;Brunborg, Gunnar
通讯作者：
Brunborg, Gunnar
影响因子：
14.9
作者：
Olsen, AK;Duale, N;Brunborg, G
通讯作者：
Brunborg, G