Establishment of a transgenic cell model for preliminary screening of chemopreventive agents.
- Author:
Yu-Juan WU
1
;
Wen-Zhi WEI
;
Xiang-Ming LI
;
Bi-Chun LI
;
Guo-Hong CHEN
Author Information
1. Animal Science and Technology College, Yangzhou University, Yangzhou 225009, China.
- Publication Type:Journal Article
- MeSH:
Antineoplastic Agents;
pharmacology;
Base Sequence;
Dose-Response Relationship, Drug;
Drug Screening Assays, Antitumor;
methods;
Enhancer Elements, Genetic;
genetics;
Gene Expression;
drug effects;
Gentamicins;
pharmacology;
Green Fluorescent Proteins;
genetics;
metabolism;
Hep G2 Cells;
Humans;
Lentinan;
pharmacology;
Microscopy, Fluorescence;
Molecular Sequence Data;
Oligonucleotides;
genetics;
Proline;
analogs & derivatives;
pharmacology;
Recombinant Fusion Proteins;
genetics;
metabolism;
Thiocarbamates;
pharmacology;
Transfection
- From:
Chinese Journal of Biotechnology
2007;23(1):85-89
- CountryChina
- Language:Chinese
-
Abstract:
To develop a GFP transgenic cell model under the transcriptional control of TK promoter adjacent to which ARE enhancer was inserted. Synthetic oligonucleotide ARE motif was annealed and purified then inserted into pTK-GFP to construct the vector of pARE-TK-GFP. The TK and ARE-TK fragments were amplified by PCR and cloned into pEGFP-N1 to reconstruct eukaryotic expression vectors of pTK-GFP/Neo and pARE-TK-GFP/Neo. They were transfected into HepG2 cells and clones resistant G418 were isolated. PDTC and Lentinan were used to induce the cell levels of GFP and the fluorescence was measured using a fluorescence plate reader. The results showed that the induced level of GFP is significantly increased and have dose-dependeny in a certain range. This findings indicated that such a cell model offered a potential platform for preliminary screening of all kinds of natural or synthetic chemopreventive agents.
