title: Development of a screening system for DNA damage and repair of potential carcinogens based on dual luciferase assay in human HepG2 cell.
abstract: 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.
authors.affiliation: Department of Toxicology, School of Public Health, Hebei Medical University, Shijiazhuang, Hebei 050017, China.
authors.forename: Longgang
authors.initials: L
authors.lastname: Fan
authors.forename: Yujie
authors.initials: Y
authors.lastname: Niu
authors.forename: Shaohui
authors.initials: S
authors.lastname: Zhang
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.forename: Huicai
authors.initials: H
authors.lastname: Guo
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
mesh_terms: D002273:Carcinogens / Q000633:toxicity*; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009152:Mutagenicity Tests / Q000379:methods*; D012680:Sensitivity and Specificity; D051742:Transcription Factor CHOP / Q000235:genetics
chemical_list: D002273:Carcinogens; C497249:DDIT3 protein, human; D051742:Transcription Factor CHOP; D008156:Luciferases