Method for screening ecdysone-inducible stable cell lines.
Ecdysone-inducible systems are useful tools to study the function of various genes in different types of mammalian cells. However, it is technically difficult to establish stable cell lines. This is partly because the conditional expression system requires the expression of two or more components driven by different kinds of promoters. In this paper, we describe the use of a luciferase reporter gene system for rapid screening of cell lines that express the ecdysone and retinoid X receptors. Using this system, two human stable colon cancer cell lines, SW480/VgRXR and HCT116/VgRXR, were successfully generated. The expression of these receptors remained high after six months of continuous culturing. A tight regulation of gene induction in SW480/VgRXR was observed using 2 microM Ponasterone A. The gene induction was rapid and persistent. Our results demonstrated the advantage of establishing cell lines that continuously express high levels of ecdysone receptor proteins.
Wakita
K
K
University of California, San Francisco, San Francisco, CA, USA.
McCormick
F
F
Tetsu
O
O
D000818:Animals; D003110:Colonic Neoplasms; D004440:Ecdysone / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics; D017930:Genes, Reporter; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured
D004440:Ecdysone; D008156:Luciferases