title: The development of GADD45α luciferase reporter assays in human cells for assessing the genotoxicity of environmental pollutants.
abstract: OBJECTIVE
  In order to assess the potential carcinogenic and genotoxic responses induced by environmental pollutants, genotoxicity test systems based on a GADD45α promoter-driven luciferase reporter in human A549 and HepG2 cells were established.
  
  
  METHODS
  Four different types of environmental toxicants including DNA alkylating agents, precarcinogenic agents, DNA cross-linking agents and non-carcinogenic agents, and three environmental samples collected from a coke oven plant were used to evaluate the test systems. After treated with the tested agents and environmental samples for 12 h, the cell viabilities and luciferase activities of the luciferase reporter cells were determined, respectively.
  
  
  RESULTS
  Methyl methanesulfonate, benzo[a]pyrene, formaldehyde and the extractable organic matter (EOM) from coke oven emissions in ambient air generally produced significant induction of relative luciferase activity in a similar dose-dependent manner in A549- and HepG2-luciferase cells. No significant increases in relative luciferase activity were observed in pyrene-treated A549- or HepG2-luciferase cells. Significant increase in relative luciferase activity was already evident after 2.5 µM benzo[a]pyrene, 5 µM formaldehyde, 0.006 µg/L bottom-EOM, 0.10 µg/L side-EOM or 0.06 µg/L top-EOM, where no cytotoxic damage was observed. Compared with the A549-luciferase cells, the tested pollutants produced higher induction of relative luciferase activity in HepG2-luciferase cells.
  
  
  CONCLUSIONS
  Therefore, the new genotoxicity test systems can detect different types of genotoxic agents and low concentrations of environmental samples. The luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the genotoxic damage of environmental pollutants and their complex mixtures.
authors.affiliation: School of Public Health, Medical College of Soochow University , Suzhou, Jiangsu , China and.
authors.forename: Lili
authors.initials: L
authors.lastname: Xin
authors.forename: Jianshu
authors.initials: J
authors.lastname: Wang
authors.forename: Yanhu
authors.initials: Y
authors.lastname: Wu
authors.forename: Sifan
authors.initials: S
authors.lastname: Guo
mesh_terms: D000477:Alkylating Agents / Q000633:toxicity; D002273:Carcinogens / Q000633:toxicity; D018797:Cell Cycle Proteins / Q000235:genetics*; D002470:Cell Survival / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004785:Environmental Pollutants / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D056945:Hep G2 Cells; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis* / Q000235:genetics; D009152:Mutagenicity Tests / Q000379:methods*; D009687:Nuclear Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D018570:Risk Assessment; D015854:Up-Regulation; D000097642:GADD45 Proteins
chemical_list: D000477:Alkylating Agents; D002273:Carcinogens; D018797:Cell Cycle Proteins; D003432:Cross-Linking Reagents; D004785:Environmental Pollutants; D008156:Luciferases; D009687:Nuclear Proteins; C439070:GADD45A protein, human; D000097642:GADD45 Proteins
keywords: Environmental complex mixtures; GADD45α; extractable organic matter; genotoxicity assessment; luciferase reporter genes