Mir 15a

The miRNA mimics (miR-24-3p, miR-101-3p, miR-148a, miR-15a, miR-504, miR-329, miR-370, miR-149, miR-320a, miR-193b, miR-544a, miR-150, miR-124-3p), miRNA control, and miRNA inhibitors for miR-24-3p were purchased from Ribobio (Ribobio, Guangzhou, China). After seeding into 6-well plates, cells were transfected with miRNA mimics at a final concentration of 100 nM using RNAiMAX Reagent (Invitrogen).

MCF10A, MCF-7, and MDA-MB-231 cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). MCF-10AT cell line was obtained from the Karmanos Cancer Institute (KCI). MCF-10A and MCF-10AT cells were cultured in DMEM/F12 containing 5% HS, 0.5 μg/ml hydrocortisone, 100 ng/ml cholera toxin, 20 ng/ml recombinant human EGF, 10 μg/ml insulin, and 1% penicillin/streptomycin. MCF-7 cells were maintained in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. MDA-MB-231 cells were cultured in L-15 medium supplemented with 10% FBS and 1% penicillin/streptomycin. All cells were grown in a humidified atmosphere containing 5% CO₂ at 37°C.
The AE was dissolved in dimethyl sulfoxide (DMSO, Sigma) and diluted with culture medium to final concentrations before treatment. In all experiments, the final concentration of DMSO in AE medium solution was less than 0.1%, which had no effect on the cells.
miR-15a and miR-16-1 mimics and inhibitors were purchased from RiboBio (Guangzhou, China). According to the manufacturer's instructions, the cell lines were transfected with 100 nM of miR-15a, miR-16-1 or negative control using Lipofectamine 2000 (Invitrogen) in 6-well plates. Transfected cells were collected 24 h after transfection. Besides, NC-treated cells or untreated cells were used as a control group of the study.