[Establishment of a transgenic cell model for preliminary screening of chemopreventive agents]

Sheng Wu Gong Cheng Xue Bao. 2007 Jan;23(1):85-9.
[Article in 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.

MeSH terms

  • 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
  • Green Fluorescent Proteins / metabolism*
  • Hep G2 Cells
  • Humans
  • Lentinan / pharmacology
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Oligonucleotides / genetics
  • Proline / analogs & derivatives
  • Proline / pharmacology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Thiocarbamates / pharmacology
  • Transfection

Substances

  • Antineoplastic Agents
  • Gentamicins
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • Thiocarbamates
  • prolinedithiocarbamate
  • Green Fluorescent Proteins
  • Lentinan
  • Proline
  • antibiotic G 418