Cytotoxicity and Apoptosis Analysis of Pyrazole Compounds

Dimethylformamide, K₂PtCl₄, KI, acetone, N-hydroksymethyl-3,5-dimethylpyrazole, 3,5-dimethylpyrazole, 3,4-dimethylpyrazole, pyrazole, 5-methylpyrazole, N-ethylpyrazole, diethyl ether, methanol, ethidium bromide, cisplatin, 3–(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Triton X-100, formaldehyde, dimethylsulfoxide (DMSO) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Stock cultures of fibroblast cells, human MCF-7 breast cancer cells, and human MDA-MB-231 breast cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA). Dulbecco’s minimal essential medium (DMEM), fetal bovine serum (FBS), CPSR1, and PBS used in a cell culture were products of Gibco (San Diego, CA, USA). Glutamine, penicillin, and streptomycin were obtained from Quality Biologicals Inc. (Gaithersburg, MD, USA). Fluorescein isothiocyanate (FITC) Annexin V Apoptosis Detection Kit II, JC-1 MitoScreen Kit, APO-Direct Kit, anti-active-caspase-3 mouse monoclonal antibody, anti-active-caspase-8 mouse monoclonal antibody, anti-active-caspase-9 mouse monoclonal antibody, FITC anti-mouse secondary antibody were from BD Pharmingen (San Diego, CA); FLICA Caspase 3 Kit, FLICA Caspase 8 Kit, FLICA Caspase 9 Kit, Hoechst 33342 (ImmunoChemistry Technologies, Bloomington, MN, USA), RNase A Solution (Promega, Madison, WI, USA). All tested compounds were dissolved in DMSO.