Emodin induces apoptosis in human hepatocellular carcinoma HepaRG cells via the mitochondrial caspase-dependent pathway

Dong, Xiaoxv; Ni, Boran; Fu, Jing; Yin, Xingbin; You, Longtai; Leng, Xin; Liang, Xiao; Ni, Jian

Cell viability assay. Cell viability was measured using the MTT assay as previously described (32) . HepaRG cells were seeded in 96-well microplates (5x10 3 cells/well) and incubated at 37˚C overnight. After incubation with emodin (20, 40 and 80 µM) for 24 and 48 h at 37˚C, 100 µl of MTT was added into each well and incubated for 4 h at 37˚C in a 5% CO 2 incubator. Subsequently, DMSO (150 µl) was added to dissolve the formazan crystals. Absorbance of the formazan solution was read at 570 nm in a microplate reader (Multiskan GO; Thermo Fisher Scientific, Inc.). For the lactate dehydrogenase (LDH) assay, the cells were cultured overnight and then treated with serial concentrations of emodin for 24 h. The LDH enzyme released from cells was quantified by an LDH assay kit according to the manufacturer's protocol. The absorbance of the supernatant was measured at 490 nm by fluorescent plate reader.