MCF7 AREc32 Cell Line
- Name Roland Wolf
- Institute University of Dundee
Tool name: MCF7 AREc32 Cell Line
Tool type: Cell Lines
Tool sub-type: Continuous
Parental cell line: MCF7
Disease: Cancer; Drug metabolism
Description: Cell line used to investigate ARE gene expression.
Background and Research Application
The stable human mammary MCF7-derived reporter cell line called AREc32 contains copies of the rat GST antioxidant response element (ARE) linked to a luciferase gene, such that the induction of the ARE results in luciferase activity. ARE is a transcriptional cis-regulatory element involved in the activation of genes coding for a number of antioxidant proteins and enzymes that work in concert to protect tissues from oxidative insults. This cell line can be used to investigate whether anti-cancer drugs can induce ARE-driven gene expression.
Research area: Cancer; Drug development; Drug development
Production details: The ARE-luciferase reporter plasmid was generated using the pGL3-promoter vector containing an SV40 promoter upstream of the firefly luciferase gene. They differ in the number of copies of ARE sequences that have been inserted, in head-to-tail orientation, through Nhe I & Xho I restrictionsites upstream of the promoter-luc+ transcriptional unit. A plasmid was made containing eight copies of the ARE (5'-GTGACAAAGCA-3', with the minimal functional sequence underlined) present in both rat GSTA2 and mouse gsta1; called pGL-8xARE. A linker with the sequence of 5'-CCC-3' and 5'-GGG-3' on the opposite strand was placed between individual cis-elements.pGL-8xARE, was stably transfected into MCF7 cells using the calcium phosphate method. Transfected cells were selected using 0.8 mg/mL G418 in the media for 3 to 4 weeks. The G418-resistant clones were isolated and screened by measuring their basal and inducible (obtained by treatment with 50 Amol/L t-BHQ) luciferase activities as described above. Positive clones, which showed low background and high inducible luciferase activity, were passaged and maintained in growth medium containing 0.8 mg/mL G418.
Cellosaurus ID: CVCL_1D32
- For Research Use Only
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