Dmem f12

Human breast cancer cell line MCF-7 and mammary epithelial cell line MCF-10A were obtained from cell resources center of Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences (Beijing, China). MCF-7 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Macgene, Beijing, China) supplemented with 10% FBS (PAN Biotech, Aidenbach, Germany) and 1% antibiotic-antimycotic solution. MCF-10A cells were cultured in phenol red-free DMEM-F12 (Macgene, Beijing, China) medium supplemented with 5% horse serum (Invitrogen, Grand Island, NY, USA), epidermal growth factor (20 ng/ml), hydrocortisone (500 ng/ml), insulin (10 μg/ml), cholera toxin (100 ng/ml) and 1% antibiotic/antimycotic mixture. Both cells were maintained at 37 °C in a humidified incubator with 5% CO₂. For chronic oxidative stress study, MCF-10A cells were exposed to H₂O₂ for 8 weeks, re-treated every 3 days and sub-cultured once a week.

The HEK-293T cell line, human mammary epithelial cell line MCF-10 A, and human breast cancer (BC) cell lines MDA-MB-231, MDA-MB-468, MCF-7, and BT549 were purchased from the Cell Bank, Type Culture Collection Committee, Chinese Academy of Sciences (Shanghai, China). Cell lines were maintained under standard media and conditions. MDA-MB-231, MDA-MB-468, MCF-7, and HEK293T were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Rockville, MD, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Rockville, MD, USA) and 1% antibiotic solution. The culture medium for BT549 was Roswell Park Memorial Institute (RPMI, Gibco)1640 medium (Gibco, Rockville, MD, USA) supplemented with 10% FBS and 1% antibiotic solution. MCF-10 A was cultured in DMEM/F12 (Macgene, Beijing, China) medium supplemented with 5% horse serum, 10 μg/ml insulin (Macgene, Beijing, China), 20 ng/ml epidermal growth factor, 100 ng/ml cholera toxin (Macgene, Beijing, China), and 0.5 μg/ml hydrocortisone (Macgene, Beijing, China). All cell lines were cultured at 37° C, 5% CO₂ in a humidified cell culture incubator.