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MCF7 AREc32 Cell Line

Invented by Prof Roland Wolf from University of Dundee
Invented at University of Dundee

Info

Catalogue Number 151014
Antigen/Gene or Protein Targets Antioxidant Response Element (ARE)- Luciferase
Parental Line MCF7
Host Human
Tissue Breast
Disease Keywords Cancer; drug metabolism
Model Reporter
Relevance 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.
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.
Research Area Cancer, Drug Discovery & Development
Recommended Growing Conditions DMEM with glutamax supplemented with 10% fetal bovine serum and antibiotics. Do not culture beyond 15 passages after revival.
Notes Concentration
Vial has between 1-5 million cells as standard, however this may vary.
Cellosaurus ID CVCL_1D32

References

There are 20 reference entries for this reagent.

View All References

References: 20 entries

Gameiro et al. 2017. Sci Rep. 7:45701. PMID: 28361919.

Discovery of the first dual GSK3β inhibitor/Nrf2 inducer. A new multitarget therapeutic strategy for Alzheimer's disease.

Europe PMC ID: 28361919

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.

Europe PMC ID: 27824809

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.

Europe PMC ID: 27527356

Bräunig 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.

Europe PMC ID: 27176940

Brack et al. 2016. Sci Total Environ. 544:1073-118. PMID: 26779957.

Effect-directed analysis supporting monitoring of aquatic environments--An in-depth overview.

Europe PMC ID: 26779957

New melatonin-cinnamate hybrids as multi-target drugs for neurodegenerative diseases: Nrf2-induction, antioxidant effect and neuroprotection.

Europe PMC ID: 26496465

Rücker 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.

Europe PMC ID: 25622264

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.

Europe PMC ID: 23032559

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.

Europe PMC ID: 18048326

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.

Europe PMC ID: 17108137


Add a reference

References: 20 entries

Gameiro et al. 2017. Sci Rep. 7:45701. PMID: 28361919.

Discovery of the first dual GSK3β 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.

Bräunig 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.

Rücker 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.


Add a reference

Inventor Information