In vitro investigations of glycidamide-induced DNA lesions in mouse male germ cells and in mouse and human lymphocytes
In vitro investigations of glycidamide-induced DNA lesions in mouse male germ cells and in mouse and human lymphocytes
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DOI：
10.1016/j.mrgentox.2009.12.012
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发表时间：
2010-02-02
影响因子：
1.9
通讯作者：
Brunborg, Gunnar
中科院分区：
文献类型：
--
作者：
Hansen, Siri Helland;Olsen, Ann Karin;Brunborg, Gunnar
工业化合物和食品污染物丙烯酰胺（AA）是一种可能的人类致癌物，也已知会诱导雄性介导的动物生殖影响。大多数数据表明，其代谢物缩水甘油胺（CA）与观察到的毒性有关。我们已经研究了AA/GA在小鼠雄性生殖细胞中的体外作用（在精子细胞之前！延伸）和人和小鼠外周血淋巴细胞，以评估易感性的种间和细胞类型差异，并探索DNA损伤的性质以及它们的潜在修复。彗星试验与DNA修复酶Fpg和hOGG 1结合使用，以测量特定的DNA损伤。与AA相反，CA在小鼠睾丸细胞和人外周血淋巴细胞（hPBL）中以毫摩尔浓度诱导显著水平的DNA损伤（链断裂和碱不稳定位点）。使用Fpg，在所有研究的细胞类型中，以20-50倍的灵敏度测量GA诱导的DNA损伤。GA诱导的DNA损伤不能被hOGG 1识别，这表明，基于这些修复酶的已知亲和力，涉及鸟嘌呤的烷基化，而不是氧化。人淋巴细胞似乎比两种类型的小鼠细胞更容易受到CA诱导的病变。小鼠睾丸细胞和淋巴细胞似乎对GA诱导的Fpg敏感的DNA损伤有类似的反应。病变的持久性进行了探讨，无论是精通或缺乏Ogg 1（小鼠8-氧代鸟嘌呤DNA糖基化酶）的小鼠细胞。在Ogg 1（+/+）和Ogg 1（-/-）小鼠的原代雄性生殖细胞和淋巴细胞中观察到GA诱导的Fpg敏感性损伤的体外修复率较低。我们的结论是，可能有小鼠和人类之间的差异，AA/GA诱导的遗传毒性，和小鼠雄性生殖细胞的DNA并没有出现更敏感的CA比DNA从外周血淋巴细胞在体外。证明了彗星试验与DNA修复酶结合的有用性。(C)2009爱思唯尔有限公司版权所有。
The industrial compound and food contaminant acrylamide (AA) is a probable human carcinogen, also known to induce male-mediated reproductive effects in animals. Most data suggest that its metabolite glycidamide (CA) is involved in the observed toxicity. We have investigated in vitro effects of AA/GA in mouse male germ cells (prior to spermatic! elongation) and human and mouse peripheral blood lymphocytes, to assess inter-species and cell-type differences in susceptibility, and to explore the nature of the DNA lesion(s) as well as their potential repair. The comet assay was used in combination with the DNA-repair enzymes Fpg and hOGG1 to measure specific DNA lesions. In contrast to AA, CA induced significant levels of DNA lesions (strand breaks and alkali-labile sites) at millimolar concentrations in mouse testicular cells and human peripheral blood lymphocytes (hPBL). Using Fpg, the GA-induced DNA damage was measured at 20-50-fold higher sensitivity, in all cell types investigated. GA-induced DNA damage could not be recognised by hOGG1, suggesting that, based on the known affinities of these repair enzymes, alkylation of guanine is involved, rather than oxidation. Human lymphocytes appeared to be more susceptible to CA-induced lesions than both types of mouse cells. Mouse testicular cells and lymphocytes seemed to respond similarly to GA-induced Fpg-sensitive DNA lesions. The persistence of lesions was explored with cells from mice either proficient or deficient in Ogg1 (mouse 8-oxoguanine DNA glycosylase). Low in vitro repair of GA-induced Fpg-sensitive lesions was observed in primary male germ cells and lymphocytes from both Ogg1(+/+) and Ogg1(-/-) mice. We conclude that there may be differences between mice and humans in AA/GA-induced genotoxicity, and DNA from mouse male germ cells does not appear to be more sensitive to CA than DNA from peripheral blood lymphocytes in vitro. The usefulness of the comet assay in combination with DNA-repair enzymes is demonstrated. (C) 2009 Elsevier B.V. All rights reserved.