Mda mb 436

The human ER and PR positive BC cell line MCF-7 (ATCC® HTB-22™), HER-2 positive BC cell line SK-BR-3 (ATCC® HTB-30™), TNBC cell lines MDA-MB-436 (ATCC® HTB-130™), BT-20 (ATCC® HTB-19™) and MDA-MB-231 (ATCC® CRM-HTB-26™), and human non-cancerous mammary epithelial cell line (MCF-10A; ATCC® CRL-10317™) were purchased from the American Type Culture Collection (Manassas, USA). MDA-MB-231-luc cell line was a gift by Dr. Bin Gao, Ph.D., Professor from Chinese Academy of Sciences. MCF-7, SK-BR-3 and BT20 cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Sigma, USA) supplemented with 10% FBS, 100 U/mL penicillin and 50 U/mL streptomycin in a humidified atmosphere containing 5% CO₂, at 37°C. MDA-MB-231 and MDA-MB-436 cells were maintained in L15 medium (Sigma, USA) supplemented with 10% FBS, 100 U/mL penicillin and 50 U/mL streptomycin in a humidified atmosphere at 37°C. MCF-10A cells were cultured in DMEM/Nutrient Mixture F-12 Ham medium (DMEM/F-12; Sigma, USA) containing 5% horse serum in the presence of 10 μg/mL insulin (Sigma, USA), 20 ng/mL epidermal growth factor (EGF; Sigma, USA), 100 ng/mL cholera toxins (Sigma, USA), 0.5 μg/mL hydrocortisone (Sigma, USA), 100 U/mL penicillin, and 50 U/mL streptomycin. All cells were used in experiments during the linear phase of growth.

The human mammary epithelial cell line MCF-10A and TNBC cell lines MDA-MB-436, MDA-MB-231, MDA-MB-468, BT-549, BT-20 and Human Embryonic Kidney 293 (HEK293) cells were purchased from the American Type Culture Collection (Manassas, VA). MDA-MB-436, MDA-MB-231, MDA-MB-468, BT-549, BT-20 and HEK293 cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 supplemented with 10% FBS and a 100-U/ml penicillin-streptomycin solution (Sigma). MCF-10A cells were maintained in a nutrient mixture consisting of DMEM/F12 supplemented with 5% horse serum, epidermal growth factor, hydrocortisone, insulin and cholera toxin. All cultured cells were incubated at 37°C in a water-saturated 95% air–5% CO₂ atmosphere.

The human cell lines of TNBC, MDA-MB-231, and MDA-MB-436 were obtained from The Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China) and cultured with Dulbecco's Modified Eagle's medium (Gibco, Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum and 1 × penicillin streptomycin (Corning) in a 37°C incubator with an atmosphere of 5% CO₂ and 95% air. MDA-MB-231 and MDA-MB-436 paclitaxel-resistant cell lines were established by gradual administration of increasing concentration of paclitaxel (Sigma-Aldrich, USA) into culture medium from 1 nM to 100 nM for 6 months. The medium with paclitaxel was changed every two days.