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http://dx.doi.org/10.7314/APJCP.2013.14.7.4347

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)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.14, no.7, 2013 , pp. 4347-4352 More about this Journal
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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1 Yamanaka K, Rocchi P, Miyake H, et al (2005). A novel antisense oligonucleotide inhibiting several antiapoptotic Bcl-2 family members induces apoptosis and enhances chemosensitivity in androgen-independent human prostate cancer PC3 cells. Mol Cancer Ther, 4, 1689-98.   DOI   ScienceOn
2 Ghosh G, van Duyne G, Ghosh S, Sigler PB (1995). Structure of $NF-{\kappa}B$ p50 homodimer bound to a kB site. Nature, 373, 303-10.   DOI   ScienceOn
3 Jemal A, Bray F, Center MM, et al (2011). Global Cancer Statistics. CA Cancer J Clin, 61, 69-90.   DOI
4 Leelavinothan P, Kalist S (2011). Beneficial effect of hesperetin on cadmium induced oxidative stress in rats: an in vivo and in vitro study. Eur Rev Med Pharmacol Sci, 15, 992-1002
5 Liu D, Huang Z (2011). Synthetic peptides and non-peptidic molecules as probes of structure and function of Bcl-2 family proteins and modulators of apoptosis. Apoptosis, 6, 453-62.
6 Lowry OH, Rosebrough NJ, Farr AL, Randall RY (1951). Protein measurement with the Folin-phenol reagent. J Biol Chem, 193, 265-75.
7 Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65, 55-63.   DOI   ScienceOn
8 Nalini N, Aranganathan S, Kabalimurthy J (2012). Chemopreventive efficacy of hesperetin (citrus flavonone) against 1,2-dimethylhydrazine-induced rat colon carcinogenesis. Toxicol Mech Methods, 22, 397-408.   DOI   ScienceOn
9 Orallo F, Alvarez E, Basaran H, Lugnier C (2004). Comparative study of the vasorelaxant activity, superoxide-scavenging ability and cyclic nucleotide phosphodiesterase-inhibitory effects of hesperitin and hesperidin. Nauyn-Schmeideberg's Arch Pharmacol, 370, 452-63.   DOI
10 Perkins ND (2012). The diverse and complex roles of $NF-{\kappa}B$ subunits in cancer. Nat Rev Cancer, 12, 121-32.
11 Ririe KM, Rasmussen RP, Wittwer CT (1997). Product differentiation by analysis of DNA melting curves during polymerase chain reaction. Anal Biochem, 245, 154-60.   DOI   ScienceOn
12 Schmittgen TD and Livak KJ (2008). Analyzing real-time PCR data by the comparative CT method. Nature Protocols, 3, 1101-8.   DOI   ScienceOn
13 So FV, Guthrie N, Chambers AF, Carroll KK (1997). Inhibition of proliferation of estrogen receptor-positive MCF-7 human reast cancer cells by flavonoids in the presence and absence of excess estrogen. Cancer Lett, 112, 127-33.   DOI   ScienceOn
14 Tanaka T, Makita H, Kawabata K, et al (1997). Chemoprevention of azoxymethane-induced rat colon carcinogenesis by the naturally occurring flavonoids, diosmin and hesperidin. Carcinogenesis, 18, 957-65.   DOI   ScienceOn
15 van de Loosdrecht AA, Beelen RH, Ossenkoppele GJ, Broekhoven MG, Langenhuijsen MM (1994). A tetrazoliumbased colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. J Immunol Methods, 174, 311-20.   DOI   ScienceOn
16 Vladic-Stjernholm Y, Vladic T, Blesson CS, Ekman-Ordeberg G, Sahlin L (2009). Prostaglandin treatment is associated with a withdrawal of progesterone and androgen at the receptor level in the uterine cervix. Reprod Biol Endocrinol, 7, 116.   DOI   ScienceOn
17 Xiao G, Fu J (2011). $NF-{\kappa}B$ and cancer: a paradigm of Yin-Yang. Am J Cancer Res, 1, 192-221.
18 Billen LP, Kokoski CL, Lovell JF, Leber B, Andrews DW (2008). Bcl-XL inhibits membrane permeabilization by competing with Bax. PLoS Biol, 6, 147.   DOI   ScienceOn
19 Choi EJ (2007). Hesperetin induced G1-phase cell cycle arrest in human breast cancer MCF-7 cells: involvement of CDK4 and p21. Nutr Cancer, 59, 115-9.   DOI   ScienceOn
20 Borradaile NM, Carroll KK, Kurowska EM (1999). Regulation of HepG2 cell apolipoprotein B metabolism by the citrus flavanones hesperetin and naringenin. Lipids, 34, 591-8.   DOI   ScienceOn
21 Choi EJ (2008). Antioxidative effects of hesperetin against 7, 12-dimethylbenz(a)anthracene-induced oxidative stress in mice. Life Sciences, 82, 1059-64.   DOI   ScienceOn
22 Dolcet X, Llobet D, Pallares J, Matias-Guiu X (2005). $NF-{\kappa}B$ in development and progression of human cancer. Virchows Arch, 446, 475-82.   DOI
23 Elmore S (2007). Apoptosis: a review of programmed cell death. Toxicol Pathol, 35, 495-516.   DOI   ScienceOn
24 Elumalai P, Gunadharini DN, Senthilkumar K, et al (2012). Induction of apoptosis in human breast cancer cells by nimbolide through extrinsic and intrinsic pathway. Toxicol Lett, 215, 131-42.   DOI   ScienceOn