Figure 5
(a) Representative images of MDA-MB-231 cells (magnification 20×). Untreated and treated MDA-MB-231 cells with 2.5 μg/mL (IC50) compound C1 and 0.9 μg/mL (IC50) cisplatin were stained with Hoechst 33342, cell membrane permeability, cytochrome c and MMP dyes. Treated MDA-MB-231 cells with compound C1 and cisplatin produced a marked decrease in MMP and a noteworthy elevation in total nuclear intensity, membrane permeability and cytochrome c. (b) Western blot analysis of cytosolic cytochrome c: Cytochrome c Releasing Apoptosis Assay Kit was used to evaluate the release of cytochrome c from the mitochondria to cytosol in untreated and treated MDA-MB-231 cells with 2.5 μg/mL (IC50) compound C. The result showed notable release of cytochrome cin cytosol after 24, 48, and 72 ours treatments. The data represent the means ± standard deviations (SDs) of 3 independent tests. Statistical analysis is defined as significant if *P < 0.05. (c) Quantitative analysis of multiparameter cytotoxicity assay in MDA-MB-231 cells: Simultaneous alterations in total nuclear intensity, cell permeability mitochondrial membrane potential, and cytochrome c release were quantified in MDA-MB-23 cells. After treatment with compound C1 (at IC50 concentration of 2.5 μg/mL), and cisplatin (at IC50 concentration of 0.9 μg/mL), statistically significant loss of mitochondrial membrane potential and an obvious increase in total nuclear intensity, cell permeability, and cytochrome c release from mitochondria were observed.
