癌变·畸变·突变Issue(6)：461-464,469,5.DOI:10.3969/j.issn.1004-616x.2013.06.013
基于双荧光素酶报告基因的DNA损伤与修复检测系统的建立与应用
Development and application of a screening system for DNA damage and repair based on dual luciferase assay in human HepG2 cells
摘要
Abstract
OBJECTIVE: Development of a screening system for DNA damage and repair based on dual luciferase assay in human HepG2 cells. METHODS:5μmol/L CdCl2 was used to treat the plasmid pTK-RL in vitro. Plasmid pgadd153-luc and damaged pTK-RL were co-transfected into HepG2 cells and then the reconstituted HepG2 cells were treated with sixteen DNA-damaging agents and three non-genotoxic agents. The DNA damage and DNA repair abilities of HepG2 cells were measured by dual luciferase assay. Three organic extracts from different sites of soil (residential area,garbage dump and farmland) were evaluated by dual luciferase assay. DNA damage was detected by Comet assay. RESULTS:The dual luciferase assay could identify genotoxic and non-genotoxic agents. The three different soil samples had various levels of inducing DNA damage and decreasing DNA repair capacities in HepG2 cells following the order:residential area>garbage dump>farmland. CONCLUSION:In this study,the dual luciferase assay was developed to measure DNA damage and repair.关键词
DNA损伤/DNA修复/宿主细胞恢复活性试验/双荧光素酶报告基因/致癌物质Key words
DNA damage/DNA repair/host cell reactivation assay/dual luciferase report gene/carcinogen分类
医药卫生引用本文复制引用
张荣,牛玉杰,樊龙刚,石磊,王茜,王秀荣..基于双荧光素酶报告基因的DNA损伤与修复检测系统的建立与应用[J].癌变·畸变·突变,2013,(6):461-464,469,5.基金项目
国家自然科学基金(81072269，81102151)，河北省自然科学基金(C2011206048) （81072269，81102151）
