Mcf 7 cells

MCF-7 cells were obtained from the China Center for Type Culture Collection (Wuhan, Hubei, China). Cells were cultured in RPMI-1640 medium (Gibco, Waltham, MA, USA) with 10% newborn bovine serum, in a humidified 5% CO₂ incubator, at 37 °C. All drugs (purity > 98%) were purchased from Haoyuan Chemexpress Co., Ltd. (Shanghai, China) and dissolved in sterile dimethyl sulfoxide (DMSO) at 100 mmol/L (gefitinib, sorafenib and everolimus) or 200 mmol/L (cladribine, dasatinib and vorinostat ) stock solutions.

C57BL/6 background B16F0 and B16F1 melanoma cells, human hepatocellular carcinoma HepG2 cells, and human breast cancer MCF-7 cells were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China) and cultured according to their guidelines. The cell lines were authenticated at China Center for Type Culture Collection (Wuhan, China) in June 2014, using short tandem repeat (STR) DNA profiling (ABI 31300xl Genetic Analyzer; Life Technologies). To induce the invasive capacity of non-metastatic tumor cells (B16F0, HepG2, MCF-7), the cells were cultured in presence of T/H/H (TGF-β1, 5 ng/ml, H₂O₂, 100 μM, HOCl, 50 μM) for 10 days [19 (link)]. For the convenience of description, T/H/H-pretreated cells were mentioned as T/H/H-B16F0, T/H/H-HepG2, and T/H/H-MCF-7 cells, respectively.

MCF-7 cells were purchased from the China Center for Type Culture Collection (Wuhan, China) and cultured in DMEM containing 10% FBS and 1% antibiotics (penicillin/streptomycin).
6-week-old female ICR mice (18–22 g of body weight) and BALB/c mice (4–5 weeks old, 14–16 g of body weight) were obtained from Shanghai Slac Laboratory Animal Co. Ltd. All animal procedures were approved by the Animal Experimental Center of Zhejiang University of Technology (Hangzhou, China) and followed the guidelines for care and use of laboratory animals.

Docetaxel was purchased from Shanghai Techwell Biopharmaceutical Co. Ltd (Shanghai, China). D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS) was purchased from Sigma–Aldrich Co. (MO, USA). LDH commercial test kit was supplied by Jiancheng Biotech Institute (Nanjing, China). Glucose commercial test kit was purchased from Rongsheng Biotech Co. Ltd (Shanghai, China). Cell counting kit-8 was from Dojindo Laboratories (Kumamoto, Japan). Dulbecco’s modified Eagle medium, fetal bovine serum, nonessential amino acid, penicillin, and streptomycin were from Invitrogen Life Technologies (Carlsbad, CA). All other chemicals and solvents were of analytical reagent grade.
Male Sprague Dawley rats of 200–220 g weight and female nude mice of 4–6 weeks were from Shanghai Super B&K Laboratory Animal Corporation Ltd (Shanghai, China) and maintained at 22 °C ± 2 °C on a 12–hour light–dark cycle with access to food and water ad libitum. The animals used for the experiment were treated according to the protocols evaluated and approved by the ethical committee of Shanghai University of Traditional Chinese Medicine (Shanghai, China). Caco-2 cells were a kind gift from Professor Jianxin Wang (School of Pharmacy, Fudan University, Shanghai, China). MCF-7 cells were obtained from Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China).

Human breast cancer MCF-7 cells were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in high-glucose Dulbecco's modified eagle medium (DMEM) (Gibco, USA) supplemented with 1% penicillin/streptomycin and 10% (v/v) (Gibco) fetal bovine serum (FBS) (Gibco, USA) and maintained at 37 °C in a 5% CO₂ incubator (Thermo, USA). Cells were seeded in 6-well plates or 24-well plates at at a density of 10⁵ cells/cm² the day before the experiments. When 70%-80% confluence was achieved, Orp (125, 250, 500, 1000, and 2000 μg/mL) were added to the plates for 24 h incubation and 20 ng/mL TNF-α were added into cell culture 30 min before the end of Orp treatment. For the treatment of inhibitor, 20 μM MG132 (Selleck, USA) was used to block the ubiquitin-proteasome pathway 6 h before the treatment of TNF-α and Orp. Following this, cytosolic protein and nuclear protein were extracted using a nuclear/cytoplasmic protein separation kit.