Potential effects of hesperidin isolated from Zanthoxylum rhetsa against NTERA-2 cancer stem cells

Abstract

Recent studies have suggested that cancer stem cells (CSCs) may represent novel targets for cancer therapies. Besides, flavonoids derived from plants have attracted interest because of their promising bioactivities and medicinal potential. Hesperidin (HES) isolated from Zanthoxylum rhetsa has been demonstrated to have potent cytotoxic effects against a category of cancer cell lines. This study firstly proposes to evaluate the bioactivity of hesperidin against NTERA-2 cancer stem cells. Hesperidin has been examined for the suppression capacities on proliferative ability and stemness properties of NTERA-2 cells. The compound was investigated potential activities via 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay in 2D, 3D culture, flow cytometry, caspase assay and western blot analysis. The results showed that hesperidin exerted anti-proliferative activity, with a half-maximal inhibitory concentration (IC₅₀) of 57.80 ± 3.55 µM against NTERA-2 cells, but showed much less toxicity against HEK-293A human embryonic kidney cells (IC₅₀> 150 µM). The anti-CSC effects of HES on NTERA-2 cells might involve in the p53 pathway, inducing apoptosis by increasing early and late apoptotic rates and activating caspase-3, -8, and -9, increasing expression of Bax, p53 proteins, and significant cell cycle arrest at the G2/M phase. Moreover, hesperidin was observed remarkably down-regulating the stemness characteristics of NTERA-2 cell by inhibiting cell proliferation in tumorsphere and suppressing the expression of Oct4, Nanog proteins. Together, these results suggest the potential of hesperidin in CSC-targeted treatment for more efficient clinical therapy.