Novel In Vivo CometChip Reveals NDMA-Induced DNA Damage and Repair in Multiple Mouse Tissues.
Novel In Vivo CometChip Reveals NDMA-Induced DNA Damage and Repair in Multiple Mouse Tissues.
复制标题
DOI：
10.3390/ijms231911776
复制
发表时间：
2022-10-04
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
--
作者：
彗星测定法是一种用于检测真核细胞DNA损伤的多功能测定法。该试验可以测量各种类型损伤的水平，包括DNA链断裂、碱基位点和碱敏感位点。此外，还可以修改该测定以包括纯化的DNA糖基化酶，以便可以检测烷基化和氧化碱基。CometChip是传统彗星试验的高通量版本，已用于研究培养细胞。在这里，我们测试了它在体内DNA损伤研究中的效用。我们发现CometChip在暴露于直接作用的甲基化剂甲基甲烷磺酸盐（MMS）和代谢激活的甲基化剂n -亚硝基二甲胺（NDMA）的小鼠的多个组织中有效检测DNA损伤，NDMA已被发现会污染食物、水和药物。具体来说，暴露于mms的小鼠的结果表明，在肝、肺、肾、胰腺、脑和脾的细胞中可以检测到DNA损伤。NDMA的结果表明，DNA损伤在代谢能力强的组织（肝、肺和肾）中可检测到，并且体内DNA修复可以随着时间的推移而监测。此外，发现DNA损伤在暴露后持续许多天。此外，糖基化酶被成功地加入到实验中，以揭示受损碱基的存在。总的来说，这项工作证明了CometChip在体内的功效，并为MMS和NDMA引起的DNA损伤的形成和修复提供了新的见解。
The comet assay is a versatile assay for detecting DNA damage in eukaryotic cells. The assay can measure the levels of various types of damage, including DNA strand breaks, abasic sites and alkali-sensitive sites. Furthermore, the assay can also be modified to include purified DNA glycosylases so that alkylated and oxidized bases can be detected. The CometChip is a higher throughput version of the traditional comet assay and has been used to study cultured cells. Here, we have tested its utility for studies of DNA damage present in vivo. We show that the CometChip is effective in detecting DNA damage in multiple tissues of mice exposed to the direct-acting methylating agent methylmethane sulfonate (MMS) and to the metabolically activated methylating agent N-nitrosodimethylamine (NDMA), which has been found to contaminate food, water, and drugs. Specifically, results from MMS-exposed mice demonstrate that DNA damage can be detected in cells from liver, lung, kidney, pancreas, brain and spleen. Results with NDMA show that DNA damage is detectable in metabolically competent tissues (liver, lung, and kidney), and that DNA repair in vivo can be monitored over time. Additionally, it was found that DNA damage persists for many days after exposure. Furthermore, glycosylases were successfully incorporated into the assay to reveal the presence of damaged bases. Overall, this work demonstrates the efficacy of the in vivo CometChip and reveals new insights into the formation and repair of DNA damage caused by MMS and NDMA.
登录
查看更多内容
影响因子：
2.7
作者：
Jackson, Petra;Pedersen, Lourdes M.;Wallin, Hakan
通讯作者：
Wallin, Hakan
DOI：
10.1016/0027-5107(90)90173-2
发表时间：
1990-07-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
BERANEK, DT
通讯作者：
BERANEK, DT
DOI：
10.1038/nrc3185
发表时间：
2012-01-12
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
通讯作者：
--
影响因子：
1.2
作者：
Ge, Jing;Prasongtanakij, Somsak;Engelward, Bevin P.
通讯作者：
Engelward, Bevin P.
前列腺癌患者的DNA修复能力降低：使用cometchip的表型方法。
影响因子：
5.2
作者：
Ortiz-Sanchez, Carmen;Encarnacion-Medina, Jarline;Park, Jong Y.;Moreno, Natasha;Ruiz-Deya, Gilberto;Matta, Jaime
通讯作者：
Matta, Jaime