Rotenone and Diallyl Disulfide–Induced Mitochondrial Apoptosis in MCF‑7 Cells: A Combination Study

Abstract

Cancer is among the most serious problems affecting the health of people across the globe. It is therefore important to come up with better treatment options that work effectively without causing any side effects. This study examines the effect of rotenone (ROT) and Diallyl disulfide (DADS) on the MCF-7 breast cancer cells through apoptosis. Cytotoxicity of the drugs was determined by measuring IC₅₀ at 48 hours using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; their interactions were estimated using the Chou-Talalay protocol. QRT-PCR was used to analyze changes in gene expression of Bax and Bcl-2, and cell distribution was quantified by flow cytometry. Both substances (IC₅₀ values of 4.62 µM for rotenone and 14.2 µM for DADS) exhibited dose-dependent cytotoxicity. The combination effects were determined by the Chou–Talalay approach, which indicated dose-dependent synergy, additivity, and antagonism at low, medium, and high doses, respectively, based on CI < 1, CI = 1, and CI > 1, where CI was the combination index. A significant change in the expression levels of Bax and Bcl-2 genes, and their ratio were observed in treatment groups. There were more apoptotic cells for rotenone than for both DADS and the combination treatment group. The percentage of cell population was as follows: rotenone (UR~69%), DADS (UR~63%), and combination treatment (moderate). In conclusion, it could be seen that while rotenone had a greater effect on the induction of apoptosis, the effectiveness was inhibited in the case of the combination treatment group.