p53 promoter-based reporter gene in vitro assays for quick assessment of agents with genotoxic potential.
The p53 promoter-based green fluorescent protein (GFP) and luciferase reporter gene assays have been established for detecting DNA damage induced by genotoxic agents. To evaluate the system, NIH3T3 cells transfected with either pHP53-GFP or pMP53-GFP construct were treated with mitomycin or 5-fluorouracil. Expression of the GFP reporter gene was significantly and specifically induced in the cells exposed to mitomycin or 5-fluorouracil. Then we treated NIH3T3 cells harboring pHP53-Luc or pMP53-Luc vector with mitomycin, 5-fluorouracil or cisplatin at various concentrations. Similarly, exposure of the cells to these agents with genotoxic potentials resulted in a dose-dependent induction in luciferase reporter gene expression. Thus, these in vitro reporter gene assays could provide an ideal system for quick assessment or screening of agents with genotoxic potential.
Li
Huaixing
H
Department of Medical Genetics, Division of Basic Medical Sciences, Second Military Medical University, Shanghai, China.
Shi
Ke
K
Chen
Ruiwen
R
He
Yan
Y
Wu
Dan
D
Sun
Shuhan
S
D000818:Animals; D002945:Cisplatin / Q000633:toxicity; D005472:Fluorouracil / Q000633:toxicity; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D016158:Genes, p53; D049452:Green Fluorescent Proteins / Q000235:genetics; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics; D051379:Mice; D016685:Mitomycin / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods*; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D016685:Mitomycin; D008156:Luciferases; D002945:Cisplatin; D005472:Fluorouracil