[Establishment of reporter gene-based cell screening model for discovering new chemopreventive agents].
OBJECTIVE
To discover new chemopreventive agents, a drug screening cell model based on reporter gene and antioxidative response element (ARE) has been established.


METHODS
Four repeas of ARE DNA binding conserved sequences were synthesized and cloned into a GFP expression vector. This construct was stably transfected into HepG2 cells in vitro. The cell model was tested with known chemopreventive agents and the effects of resveratrol, protocatechuic aldehyde, ursolic acid and oleanolic acid at different concentration (0, 12.5, 25, 50, 100, 200 micromol x L(-1)) were observed by determining reporter gene GFP activity.


RESULTS
The induce level of GFP was regulated by ARE and the dose-dependence in a certain range was observed. The induce level of GFP by resveratrol was significantly increased.


CONCLUSIONS
The method can be used to screening of chemopreventive agents from Chinese traditional medicine by measurement of luminescent value of expressed GFP in wells of microtiter plate.
Xu
Hairong
H
Institute of Integrated Chinese and Western Medicine, Medical College of Yangzhou University, Yangzhou 225001, China.
Bu
Ping
P
Li
Xiangming
X
D000970:Antineoplastic Agents / Q000494:pharmacology*; D018890:Chemoprevention; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008954:Models, Biological; D009369:Neoplasms / Q000188:drug therapy / Q000517:prevention & control*
D000970:Antineoplastic Agents; D004365:Drugs, Chinese Herbal; D049452:Green Fluorescent Proteins