Hesperidin inhibits insulin-induced phosphoinositide 3-kinase/Akt activation in human pre-B cell line NALM-6

J Cancer Res Ther. 2018 Apr-Jun;14(3):503-508. doi: 10.4103/0973-1482.157323.


Context: It has been shown that hesperidin induces apoptosis in NALM-6 cells through inhibition of nuclear factor-kappa B (NF-κB) activation.

Aims: To investigate the effect of hesperidin on inhibition of NF-κB activation through blocking phosphoinositide 3-kinase (PI3K)/Akt pathway as a main target in cancer treatment, in NALM-6 cells.

Materials and methods: NALM-6 cells were incubated with two concentrations of hesperidin (25, 50 μM) in the presence or absence of insulin (100 nM), as a potent activator of Akt. The cytotoxic activity of hesperidin was determined by 3-(4,5-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptotic death was measured by ELISA test using cell death detection ELISA Plus kit. To assay the effect of hesperidin on Akt pathway, the phosphorylation levels of Akt, inhibitor of kappa B alpha (IκBα), and glycogen synthase kinase-3 beta (GSK-3β) and expression level of IκB kinase alpha (IKKα) were determined by Western blot analysis.

Results: Hesperidin (both concentrations) significantly reduced cells survival in the presence and absence of insulin compared to untreated cells in a time-dependent manner (P < 0.05). Hesperidin also significantly increased apoptosis in NALM-6 cells even in hyperinsulinemia condition (P < 0.0001). Hesperidin inhibited insulin-induced phosphorylation and activation of Akt, IκBα, and GSK-3β and decreased expression of IKKα.

Conclusion: The results of this study demonstrated that cytotoxic and proapoptotic actions of hesperidin are partly mediated through the suppression of PI3K3/Akt/IKK signaling pathway. So, hesperidin might act as a chemotherapeutic agent by targeting cell survival pathways.

Keywords: Apoptosis; glycogen synthase kinase-3 beta; hesperidin; inhibitor of kappa B; inhibitor of kappa B kinase; nuclear factor-kappa B; phosphoinositide 3–kinase/Akt.

MeSH terms

  • Antineoplastic Agents / pharmacology
  • Apoptosis / drug effects*
  • Cell Survival / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Glycogen Synthase Kinase 3 beta / genetics
  • Hesperidin / pharmacology*
  • Humans
  • Insulin / genetics*
  • Insulin / metabolism
  • NF-KappaB Inhibitor alpha / genetics
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics
  • Neoplasms / drug therapy*
  • Neoplasms / genetics
  • Neoplasms / pathology
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphorylation / drug effects
  • Phosphorylation / genetics
  • Precursor Cells, B-Lymphoid / drug effects
  • Precursor Cells, B-Lymphoid / pathology
  • Proto-Oncogene Proteins c-akt / genetics
  • Signal Transduction / drug effects


  • Antineoplastic Agents
  • Insulin
  • NF-kappa B
  • NF-KappaB Inhibitor alpha
  • Hesperidin
  • Phosphatidylinositol 3-Kinases
  • Glycogen Synthase Kinase 3 beta
  • Proto-Oncogene Proteins c-akt