Development of a screening system for DNA damage and repair of potential carcinogens based on dual luciferase assay in human HepG2 cell.

Development of a screening system for DNA damage and repair of potential carcinogens based on dual luciferase assay in human HepG2 cell.
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开发基于人 HepG2 细胞双荧光素酶检测的 DNA 损伤和潜在致癌物修复筛选系统。

DOI:
10.1093/mutage/get028
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发表时间:
2013-09
期刊:
影响因子:
2.7
通讯作者:
Zhang, Rong
Zhang, Rong
中科院分区:
医学4区
文献类型:
--
作者:
Fan, Longgang;Niu, Yujie;Zhang, Shaohui;Shi, Lei;Guo, Huicai;Liu, Yi;Zhang, Rong

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目前,人们使用不同的方法来检测环境中DNA损伤剂的早期生物效应。一些敏感的测试方法采用DNA损伤诱导基因RNR3、RAD51、RAD54或生长停滞和DNA损伤诱导基因153(Gadd 153)来检测DNA损伤。宿主细胞再激活 (HCR) 测定是一种功能测定,基于用受损或未受损的质粒 DNA 独立转染细胞,并可鉴定导致 DNA 修复缺陷综合征的基因。在本研究中,我们结合 gadd153-luc 测试系统和 HCR 测定,通过双荧光素酶测定来测量 DNA 损伤和 DNA 修复。我们使用了 16 种 DNA 损伤剂,所有这些都通过阳性双荧光素酶报告基因测试系统检测到。双荧光素酶测定系统检测 DNA 损伤/修复的灵敏度与 gadd153-luc 测试系统和/或 HCR 测定相同。由于 DNA 修复对于维持遗传稳定性很重要,因此 DNA 损伤和修复一直是 DNA 损伤剂早期生物效应的良好生物标志物。因此,DNA修复能力的测量应该是分子流行病学研究中的一个有价值的工具。本研究描述的双荧光素酶测定快速、方便、稳定、标准。
At present, different methods are used for the detection of early biological effects of DNA-damaging agents in environment. Some sensitive testing methods employing DNA damage-inducing genes RNR3, RAD51, RAD54 or growth-arrested and DNA damage-inducible gene 153 (Gadd 153) are used to detect the DNA damage. The host cell reactivation (HCR) assay is a functional assay that is based on the independent transfection of cells with either damaged or undamaged plasmid DNA and allows the identification of the genes responsible for DNA repair-deficient syndromes. In this study, we combined the gadd153-luc test system and HCR assay to measure the DNA damage and DNA repair by dual luciferase assay. We used 16 DNA-damaging agents all of which were detected by a positive dual luciferase reporter test system. The sensitivity of the dual luciferase assay system to detect DNA damage/repair was same as the gadd153-luc test system and/or the HCR assay. Since DNA repair is important to maintain genetic stability, DNA damage and repair have been good biomarkers of early biological effects of DNA-damaging agents. Accordingly, the measurement of DNA repair capacity should be a valued tool in molecular epidemiology studies. The dual luciferase assay described in this study is rapid, convenient, stable and standard.
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