Ceramide generation as a novel biological mechanism for chemo-preventive and cytotoxic effects of hesperidin on HT-144 melanoma cells

Abstract

IntroductionHesperidin and its aglycone, hesperetin are abundant natural flavonoid in Citrus and known for their anti-neoplastic and pro-apoptotic properties. Ceramide is a key regulator in apoptosis and cytotoxicity and its metabolizing enzymes have considered as a target in cancer studies. Acid ceramidase (ACDase) and Neutral sphingomyelinase (NSMase) are the main enzymes in ceramide metabolism. MethodsIn the present study, we investigated the effects of hesperidin and hesperetin on HT-144 human melanoma cells. The investigated cells treated with seven different concentrations of hesperidin and hesperetin (0–210 µM) and cell viability, caspase-3 and 9 activities, total cell ceramide and the activities of ACDase and NSMase were evaluated with fluorescence and spectrophotometry analysis. ResultsThe data revealed that hesperidin and hesperetin were induced cytotoxicity and activities of caspase-3 and 9 in a dose-dependent fashion (P < 0.05). Total cell ceramide and the activity of NSMase, the enzyme which elevates ceramide levels, increased and the activity of ceramide removing enzyme (ACDase) decreased efficiently by studied compounds. Hesperetin was a stronger ceramide enhancer than hesperidin. ConclusionConsequently, this study reveals new hesperidin effects on ceramide metabolism and may offer a novel and complementary tool in human melanoma therapy.