Evidence for Hesperidin as an Effective Factor in Initiating the Intrinsic Pathway of Apoptosis in KG۱aLeukemia Cells

Acute myeloid leukemia (AML) is the most common subtype of leukemia, accounting for ۶۲% of all leukemia fatalities. As a polyphenol glycoside, hesperidin triggers the apoptotic pathway, which might positively affect combating cancer cells. In this study, we investigated the pro-apoptotic effects of hesperidin in KG۱a cells. The MTT assay was used to determine the IC۵۰ of hesperidin in KG۱a cell lines. For the apoptotic cell morphology study, we used Hoechst ۳۳ ۲۵۸ staining. Activation of the caspase-۳ enzyme was evaluated by the caspase-۳ assay and spectrophotometry. Cell cycle distribution was analyzed by propidium iodide staining and flow cytometry. Moreover, p۲۱, survivin, Bax, and Bcl۲ gene expression was investigated by real- time PCR. Hesperidin decreased the viability of KG۱a leukemic cell۴s, but not that of HFF۲, a non-cancer cell line. Apoptotic cell morphological alterations and increase in caspase-۳ activity were observed after hesperidin treatment. Our results revealed that the expression of anti-apoptotic genes survivin and Bcl۲ significantly decreased with hesperidin treatment, and pro-apoptotic gene Bax and cell cycle regulator p۲۱ increased compared to the control group. These findings revealed that hesperidin may be an effective factor in initiating the intrinsic pathway of apoptosis and may be good candidate for the treatment of AML.

Keywords:

acute myeloid leukemia , hesperidin , apoptosis , cell cycle