Mcf 7

The cancer cell lines used in this study were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). They consisted of MDA-MB-231, MDA-MB-436 (ER-, PR- and HER2-), and MCF-7 (ER+, PR+ and HER2+) breast cancer cells, and the MCF-10A cell line (nontumorigenic epithelial cell line) was included to represent normal cells.
MDA-MB-231 and MCF-7 were grown in high-glucose DMEM supplemented with 5% fetal bovine serum (FBS, Biowest, Miami, FL, USA), while MDA-MB-436 cells were grown in DMEM/F12 supplemented with 10% FBS (Biowest, Miami, FL, USA).
MCF-10A cells were cultured in DMEM/F-12 supplemented with 5% horse serum (Biowest, Miami, FL, USA), 20 ng/mL epidermal growth factor (Upstate Biotechnology Incorporated, Lake Placid, NY, USA), 10 µg/mL insulin (Biofluids, Rockville, MD, USA), and 500 ng/mL hydrocortisone.
All media (Biowest, Miami, FL, USA) were also supplemented with 2 mM glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin. The cells were maintained in a humidified incubator at 37 °C with 5% CO₂.

Gefitinib, lapatinib, terazosin, alfuzosin, prazosin, irinotecan, and quinacrine were purchased from Sigma Chemical (St. Louis, MO, USA). The breast cancer cell lines used in this study, MCF-7 and MDA-MB-231, were obtained from the American Type Tissue Culture Collection (ATCC), Rockville, MD, USA. MCF-7 and MDA-MB-231 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) high glucose with phenol red, and the culture medium was supplemented with 10% fetal bovine serum (FBS) (BioWest, Riverside, MO, USA) and 1% penicillin/streptomycin as an antibiotic. The cells were incubated in culture flasks (75 cm³) at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air, all of which were carried out under sterile conditions using a laminar flow hood.

The normal human mammary epithelial cells (MCF-10A), human breast cancer cells (MCF-7), human ovarian cancer cells (A278), and human ovarian fibroblasts (HOF) healthy cell lines were purchased from American Type Culture Collection (ATCC). MCF-7 was grown in the Dulbecco’s Modified Eagle’s Medium-DMEM (BioWest) containing 1% mixture of penicillin and streptomycin (BioWest), 1% of L-glutamine (BioWest) and 10% fetal bovine serum (FBS, Gibco). To cultivate MCF-10A the Dulbecco's Modified Eagle Medium, nutrient mixture F-12 was used. The medium was supplemented with 5% horse serum (Sigma-Aldrich), 10 ng·mL⁻¹ epithelial growth factor (Sigma-Aldrich), 5 μg·mL⁻¹ hydrocortisone (Sigma-Aldrich) and 10 μg·mL⁻¹ human insulin (Sigma-Aldrich). All cell cultures were mycoplasma free. To cultivate the A2780 cell lines the RPMI 1640 (BioWest) containing 1% mixture of penicillin and streptomycin (BioWest), 10% fetal bovine serum (FBS, Gibco) and 1% of L-glutamine (BioWest) was used. The HOF cells were cultivated by fibroblast medium that consisted of 500 ml of basal medium, 10 ml of fetal bovine serum (FBS), 5 ml of fibroblasts growth supplement (FGS), and 5 ml of antibiotic solution (P/S).