MTT assay for in vitro cytotoxicity

Growth inhibitory effect toward cancer cells line was determined by means of MTT colorimetric assay (Gerlier. and Thomasset, 1986). The tumor cells were seeded in 96-well plates at 5 × 10³ cells per well. The tumor HepG2, Hela, MDAMB231 cells were treated with 10 μM–150 μM of the complex using 5-Fu as the positive control for 24 h, respectively. After incubation, culture medium was removed and 20 μL of MTT reagent was added to each well. The formazan product was dissolved by addition of 100 μL DMSO per well. Optical density (OD) values of the purple solution that represented cell viability were measured at 570 nm. The cell viability was evaluated by measuring at a wavelength of 490 nm (Spectra Max, United States) with a microplate reader.