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

  • Sambantham, Shanmugam (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Radha, Mahendran (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Paramasivam, Arumugam (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Anandan, Balakrishnan (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Malathi, Ragunathan (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Chandra, Samuel Rajkumar (Department of Genetics, Dr.ALMPGIBMS, University of Madras) ;
  • Jayaraman, Gopalswamy (Department of Genetics, Dr.ALMPGIBMS, University of Madras)
  • Published : 2013.07.30

Abstract

Aim: To investigate the molecular mechanisms underlying triggering of apoptosis by hesperetin using in silico and in vitro methods. Methods: The mechanism of binding of hesperetin with NF-${\kappa}B$ and other apoptotic proteins like BAX, BAD, $BCL_2$ and $BCL_{XL}$ was analysed in silico using Schrodinger suite 2009. In vitro studies were also carried out to evaluate the potency of hesperetin in inducing apoptosis using the human prostate cancer PC-3 cell line. Results: Hesperetin was found to exhibit high-affinity binding resulting from greater intermolecular forces between the ligand and its receptor NF-${\kappa}B$ (-7.48 Glide score). In vitro analysis using MTT assay confirmed that hesperetin reduced cell proliferation ($IC_{50}$ values of 90 and $40{\mu}M$ at 24 and 48h respectively) in PC-3 cells. Hesperetin also downregulated expression of the anti-apoptotic gene $BCL_{XL}$ at both mRNA and protein levels and increased the expression of pro-apoptotic genes like BAD at mRNA level and BAX at mRNA as well as protein levels. Conclusion: The results suggest that hesperetin can induce apoptosis by inhibiting NF-${\kappa}B$.

Keywords

Docking;hesperetin;apoptosis;NF-${\kappa}B$

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