Mda mb 435 cells

Human melanoma MDA-MB-435 cells (ATCC, Manassas, VA, USA) were maintained in RPMI 1640 medium supplemented with 10% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin. Exponentially grown cells were seeded at the density of 5000 cells/well into 96-well plates (Corning, Corning, NY, USA) and incubated overnight. Compounds dissolved in DMSO were added to achieve the specified final concentrations (total volume: 100 μL, DMSO: 0.5%). The incubation continued for 30 min at 37 °C and the HDAC activity determined using a commercial luminescent assay (HDAC-Glo™, Promega, Madison, WI, USA). The HDAC inhibitors trichostatin A (TSA, 1 nM) and SAHA (100 nM) were used as positive controls and the data presented as percentage inhibition of the solvent control.

HCT 116 cells (colorectal carcinoma), MDA-MB-435 cells (melanoma) and MCF-7 cells (breast cancer) were purchased from American Type Culture Collection, Rockville, MD, USA and grown in RPMI medium supplemented with L-glutamine (2 mM), 10% fetal bovine serum (FBS), penicillin (100 U/mL), streptomycin (100 μg/mL) and gentamicin (5 μg/mL). Cells were maintained in log phase by seeding twice a week at a density of 3 × 10⁸ cells/L in humidified 5% CO₂ atmosphere, at 37 °C. In all experiments, cells were made quiescent through overnight incubation before the treatment with the compounds or vehicle alone (control cells) No differences were found between cells treated with DMSO 0.1% and untreated cells in terms of cell number and viability.

Human leukemia CCRF-CEM cells, B cell lymphoma Raji cells, histiocytic lymphoma U937 cells, and breast cancer MDA-MB-435 cells and mouse lymphoma 2PK-3 cells were purchased (American Type Culture Collection, Manassas, VA). Characterization of cell lines was performed (RADIL, Columbia, MO) including screening for microbial contamination or mammalian inter-species contamination. Evaluation of alleles for nine genetic markers confirmed that banked cells matched previously reported genetic profiles. Cells were grown in RPMI 1640 or DMEM supplemented with 10% (v/v) FBS, passaged twice weekly for <2 months, and maintained in an incubator at 37°C with 5% CO₂.