MCF7 (MCF7L, originally from Dr. Marc Lippman’s lab) and T47D breast cancer cells were grown in RPMI-1640 medium (Lonza, Walkersville, MD) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin and glutamine. Parental MCF7 and T47D (have already become resistant to estrogen deprivation (ED)) cells were continuously treated with tamoxifen (Tam, 10−7 M, >6 months), and the resistant derivatives (TamRes) were selected when the initially sensitive cells resumed comparable growth to the parental cells. The TamRes cell lines were maintained as previously described (5). Lapatinib (GlaxoSmithKline, Research Triangle Park, NC), AZD8931, or gefitinib (AstraZeneca, UK) were dissolved in DMSO and added to cell culture at 1 μM concentration. 17β estradiol (E2), 4-hydroxy tamoxifen (for all in vitro studies) from Sigma (St Louis, MO) and ICI 182,780 (Fulvestrant, AstraZeneca, UK), were dissolved in ethanol. Tamoxifen citrate (Sigma) was used in all in vivo treatments as previously described [5 (link)].
Protocol detail
Endocrine-Resistant Breast Cancer Cell Lines
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17β estradiol
4 hydroxy tamoxifen
Azd8931
Breast cancer
Cell culture
Cell lines
Cells
Derivatives
Dmso
Estrogen
Ethanol
Fetal bovine serum
Fulvestrant
Gefitinib
Glutamine
Ici 182 780
Lapatinib
Marc
Mcf7
Parental
Penicillin
Streptomycin
Tamoxifen citrate
Corresponding Organization :
Other organizations : Baylor College of Medicine, AstraZeneca (United Kingdom)
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Protocol cited in 7 other protocols
Variable analysis
independent variables
- Tamoxifen (Tam, 10^-7 M, >6 months)
- Lapatinib (1 μM)
- AZD8931 (1 μM)
- Gefitinib (1 μM)
- 17β estradiol (E2)
- 4-hydroxy tamoxifen
- ICI 182,780 (Fulvestrant)
- Tamoxifen citrate
- Cell growth
- Cell viability
- Protein expression
- Signaling pathways
- RPMI-1640 medium (Lonza, Walkersville, MD) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin and glutamine
- Parental MCF7 and T47D cells
- TamRes cell lines
- Parental MCF7 and T47D cells
- TamRes cell lines
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