Open Accessp53 Promoter‐based Reporter Gene in vitro Assays for Quick Assessment of Agents with Genotoxic Potential
Author(s) -
LI Huaixing,
SHI Ke,
CHEN Ruiwen,
HE Yan,
WU Dan,
SUN Shuhan
Publication year - 2007
Publication title -
acta biochimica et biophysica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.771
H-Index - 57
eISSN - 1745-7270
pISSN - 1672-9145
DOI - 10.1111/j.1745-7270.2007.00269.x
Subject(s) - microbiology and biotechnology , reporter gene , green fluorescent protein , transfection , mitomycin c , luciferase , biology , in vitro , gene , gene expression , chemistry , biochemistry , genetics
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.