[Establishment of a transgenic cell model for preliminary screening of chemopreventive agents].
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.
Wu
Yu-Juan
YJ
Animal Science and Technology College, Yangzhou University, Yangzhou 225009, China.
Wei
Wen-Zhi
WZ
Li
Xiang-Ming
XM
Li
Bi-Chun
BC
Chen
Guo-Hong
GH
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001483:Base Sequence; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D007912:Lentinan / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013859:Thiocarbamates / Q000494:pharmacology; D014162:Transfection
D000970:Antineoplastic Agents; D005839:Gentamicins; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D013859:Thiocarbamates; C066229:prolinedithiocarbamate; D049452:Green Fluorescent Proteins; D007912:Lentinan; D011392:Proline; C010680:antibiotic G 418