The polyphenolic component of citrus fruits,
hesperetin (Hst), is a metabolite of
hesperidin. In this study, we examined the effect of varying doses and exposure times of
hesperetin on MCF-7 and MDA-MB-231
cancer cells, as well as MCF-10A normal cells. By using MTT assay, real-time PCR, western blot, and flow cytometry, we determined the effects of Hst on cell viability, ROS levels, and markers of cell death. Furthermore, molecular docking was used to identify Hst targets that might be involved in ROS-dependent cell death. According to the results, different concentrations of Hst induced different modes of cell death at specific ROS levels. Paraptosis occurred in all cell lines at concentration ranges of IC35 to IC60, and apoptosis occurred at concentrations greater than IC65. In addition, MDA-MB-231 cells were subjected to senescence at sub-toxic doses when treated for a long period of time. When Hst levels were higher,
N-acetylcysteine (NAC)'s effect on neutralizing ROS was more pronounced. According to the docking results, Hst may interact with several
proteins involved in the regulation of ROS. As an example, the interaction of CCS (
Copper chaperone for
superoxide dismutase) with Hst might interfere with its chaperone function in folding SOD-1 (
superoxide dismutase enzyme), contributing to an increase in cytoplasmic ROS levels. Finally, depending on the ROS level, Hst induces various modes of cell death.