Product Image

Contributor Information

  • Name Roland Wolf
  • Institute University of Dundee

Tool Details

  • Tool name: MCF7 AREc32 Cell Line
  • Tool type: Cell Lines
  • Tool sub-type: Continuous
  • Parental cell line: MCF7
  • Organism: Human
  • Tissue: Breast
  • Disease: Cancer; drug metabolism
  • Model: Reporter
  • Conditional: Yes
  • 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 Discovery & Development ; Drug metabolism
  • 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

Target Details

  • Target: Antioxidant Response Element (ARE)- Luciferase

Application Details

Handling

  • Format: Frozen
  • Growth medium: DMEM with glutamax supplemented with 10% fetal bovine serum and antibiotics. Do not culture beyond 15 passages after revival.
  • Storage conditions: Liquid Nitrogen
  • Shipping conditions: Dry ice
  • Mycoplasma free: Yes
  • Biosafety level: 1

Documentation

  • Available on request

References

  •   Gameiro et al. 2017. Sci Rep. 7:45701. PMID: 28361919.
  •   Discovery of the first dual GSK3? inhibitor/Nrf2 inducer. A new multitarget therapeutic strategy for Alzheimer's disease.
  •   MacLeod et al. 2016. Br J Cancer. :. PMID: 27824809.
  •   Aldo-keto reductases are biomarkers of NRF2 activity and are co-ordinately overexpressed in non-small cell lung cancer.
  •   Basar et al. 2016. Phytochem Anal. 27(5):233-8. PMID: 27527356.
  •   Utilization of the Ability to Induce Activation of the Nuclear Factor (Erythroid-derived 2)-like Factor 2 (Nrf2) to Assess Potential Cancer Chemopreventive Activity of Liquorice Samples.
  •   Brunig et al. 2016. Chemosphere. 156:181-90. PMID: 27176940.
  •   Bioanalytical effect-balance model to determine the bioavailability of organic contaminants in sediments affected by black and natural carbon.
  •   Brack et al. 2016. Sci Total Environ. 544:1073-118. PMID: 26779957.
  •   Effect-directed analysis supporting monitoring of aquatic environments--An in-depth overview.
  •   New melatonin-cinnamate hybrids as multi-target drugs for neurodegenerative diseases: Nrf2-induction, antioxidant effect and neuroprotection.
  •   Rcker et al. 2015. Org Biomol Chem. 13(10):3040-7. PMID: 25622264.
  •   Buendia et al. 2015. Future Med Chem. 7(15):1961-9. PMID: 26496465.
  •   Enhancing the anti-inflammatory activity of chalcones by tuning the Michael acceptor site.
  •   Escher et al. 2012. J Environ Monit. 14(11):2877-85. PMID: 23032559.
  •   Water quality assessment using the AREc32 reporter gene assay indicative of the oxidative stress response pathway.
  •   Wang et al. 2007. Proc Natl Acad Sci U S A. 104(49):19589-94. PMID: 18048326.
  •   Identification of retinoic acid as an inhibitor of transcription factor Nrf2 through activation of retinoic acid receptor alpha.
  •   Wang et al. 2006. Cancer Res. 66(22):10983-94. PMID: 17108137.
  •   Generation of a stable antioxidant response element-driven reporter gene cell line and its use to show redox-dependent activation of nrf2 by cancer chemotherapeutic agents.