Molecular Mechanism Underlying Hesperetin-induced Apoptosis by in silico Analysis and in Prostate Cancer PC-3 Cells

Aim: To investigate the molecular mechanisms underlying triggering of apoptosis by hesperetin using in silicoand in vitro methods. Methods: The mechanism of binding of hesperetin with NF-kB and other apoptotic proteinslike BAX, BAD, BCL2 and BCLXL was analysed in silico using Schrodinger suite 2009. In vitro studies were alsocarried out to evaluate the potency of hesperetin in inducing apoptosis using the human prostate cancer PC-3cell line. Results: Hesperetin was found to exhibit high-affinity binding resulting from greater intermolecularforces between the ligand and its receptor NF-kB (-7.48 Glide score). In vitro analysis using MTT assay confirmedthat hesperetin reduced cell proliferation (IC50 values of 90 and 40μM at 24 and 48h respectively) in PC-3 cells.Hesperetin also downregulated expression of the anti-apoptotic gene BCLXL at both mRNA and protein levels andincreased the expression of pro-apoptotic genes like BAD at mRNA level and BAX at mRNA as well as proteinlevels. Conclusion: The results suggest that hesperetin can induce apoptosis by inhibiting NF-kB.