Mcf 7 human breast cancer cells

KG-1, MOLM-13, and K-562 human leukemia cell lines; WM-266–4 human melanoma cancer cells; and MCF-7 human breast cancer cells were obtained from ATCC (USA). Cells were grown in RPMI 1640 GlutaMAX (ThermoFisher Scientific) medium containing 10% FCS (Lonza) for K-562 and MCF-7 cells and 20% for KG-1 and MOLM-13 cells, the WM-266–4 cell line was grown in EMEM containing 10% FCS at 37 °C and under 5% CO₂.

MCF-7 human breast cancer cells were obtained from ATCC (Rockville, MD) and routinely maintained in DMEM containing 10% FBS. The cells were trypsinized and plated at a density of 4×10⁵ cells/well in a 6-well plate in the complete medium and allowed to recover. After 2 days, the cells were washed with phosphate-buffered saline (PBS), and serum-starved in DMEM, without FBS, for 2 days. The serum-starved cells were treated with Cd in DMEM. After treatment, the medium was removed and the cells were washed with ice-cold PBS and lysed with 150 μl ice-cold RIPA buffer (150 mM sodium chloride, 100 mM Tris-HCl pH7.5, 1% deoxycholate, 0.1% SDS, 1% Triton X-100) supplemented with 2 mM phenylmethylsulphonyl fluoride, protease inhibitor cocktail, and phosphatase inhibitor cocktail. The cell lysates were inactivated at 95°C for 3 min, sonicated for 15 sec, and heated again at 95°C for 5 min in the presence of 5% mercaptoethanol. Protein concentration was determined by the micro BCA kit from Pierce, Thermoscientific (Rockford, IL).

LNCaP human prostate cancer cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in RPMI 1640 medium (Thermo Fisher, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA), 100 U mL⁻¹ penicillin and 100 μg mL⁻¹ streptomycin (1% P/S, Thermo Fisher, Waltham, MA, USA). MCF-7 human breast cancer cells were obtained from ATCC and maintained in high glucose Dulbecco’s Minimum Essential Medium (DMEM, Thermo Fisher, Waltham, MA, USA) supplemented with 10% FBS and 1% P/S. Cell lines were cultured according to the manufacturer’s protocol and were grown until 70% confluence before device seeding.

Dulbecco's Modified Eagle Medium/High glucose (DMEM) with L-glutamine and sodium pyruvate, phenol red-free DMEM, phosphate buffered saline (PBS), fetal bovine serum (FBS), charcoal-treated FBS, penicillin, and streptomycin were purchased from Thermo Fisher Scientific (Pittsburgh, PA). Sodium dodecyl sulfate (SDS), 30 % acrylamide/bis solution, ammonium persulfate, Tween-20, and 2-mercaptoethanol was obtained from Bio-RAD (Hercules, CA). Non-specific control RNA (cRNAi) (cat # D-001810-01-20), short interfering RNA (siRNA) against AHR (AHR-siRNA, cat # J-004990-08-0010), RELA (RELA-siRNA, cat # J-003533-06-0010) and DharmaFECT 1 Transfection Reagent (#1) were purchased from GE Healthcare Life Sciences (Pittsburgh, PA). 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) was obtained from Cambridge Isotopes Laboratory (Andover, MA). MCF-7 human breast cancer cells were purchased from ATCC (Manassas, VA) and maintained in DMEM, 10% FBS, with penicillin (100 IU/mL) and streptomycin 100 (µg/mL).