Investigational New Drugs

, Volume 28, Issue 5, pp 624–633

Involvement of non-protein thiols, mitochondrial dysfunction, reactive oxygen species and p53 in honey-induced apoptosis

PRECLINICAL STUDIES

Summary

Honey is a complex mixture of different biologically active constituents. Honey possesses anti-inflammatory, antioxidant and antitumor properties. Our chief investigation was to assess the honey induced apoptosis and its molecular mechanism in colon cancer cell growth inhibition. Honey exerted antiproliferative potential against the HCT-15 and HT-29 colon cancer cells as assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Flow cytometric analysis showed the increasing accumulation of hypodiploid nuclei in the sub-G1 phase of cell cycle indicating apoptosis. Honey transduced the apoptotic signal via initial depletion of intracellular non protein thiols, consequently reducing the mitochondrial membrane potential (MMP) and increasing the reactive oxygen species (ROS) generation. An increasing earlier lipid layer break was observed in the treated cells compared to the control. Honey induced apoptosis was accompanied by up-regulating the p53 and modulating the expression of pro and anti-apoptotic proteins. Further apoptosis induction was substantiated using DNA fragmentation assay and YO-PRO-1 staining. Results showed honey as a plausible candidate for induction of apoptosis through ROS and mitochondria-dependent mechanisms in colon cancer cells. This will promote honey as a potential chemotherapeutic agent against colon cancer.

Keywords

Honey Thiol ROS Apoptosis Mitochondrial dysfunction p53 YOPRO-1 DNA fragmentation Colon cancer 

References

  1. 1.
  2. 2.
    National cancer institute (2009) http://seer.cancer.gov/statfacts/html/colorect.html accessed on 03 Aug 2009
  3. 3.
    Wollgast J, Anklam E (2000) Review on polyphenols in Theobroma cacao: changes in composition during the manufacture of chocolate and methodology for identification and quantification. Food Res Int 33:423–447CrossRefGoogle Scholar
  4. 4.
    Madhavi DV, Despande SS, Salunkhe DK (1996) Food antioxidants. Dekker, New YorkGoogle Scholar
  5. 5.
    Chinthalapally V, Dhimant D, Barbara A et al (1993) Inhibitory effect of caffeic acid esters on azoxymethane-induced biochemical changes and aberrant crypt foci formation in rat colon. Cancer Res 53:4182–4188Google Scholar
  6. 6.
    Gribel NV, Pashiniski VG (1990) Antitumor properties of honey. Vopr Onkol 36:704–707PubMedGoogle Scholar
  7. 7.
    Tarek S, Naoto M, Mizuki O et al (2003) Antineoplastic activity of honey in an experimental bladder cancer implantation model: in vivo and in vitro studies. Int J Urol 10:213–219CrossRefGoogle Scholar
  8. 8.
    Orsolic N, Knezevi A et al (2004) Influence of honey bee products on transplantable tumours. Vet Comp Oncology 1:216–226CrossRefGoogle Scholar
  9. 9.
    Earnshaw WC (1995) Nuclear changes in apoptosis. Curr Opin Cell Biol 7:337–343CrossRefPubMedGoogle Scholar
  10. 10.
    Lee H, Wei Y (2000) Mitochondrial role in life and death of the cell. J Biomed Sci 7:2–15CrossRefPubMedGoogle Scholar
  11. 11.
    Michael OH (2000) The biochemistry of apoptosis. Nature 407:770–776CrossRefGoogle Scholar
  12. 12.
    Jaganathan SK, Mahitosh M (2009) Honey constituents and its apoptotic effect in colon cancer cells. JAAS 1:29–36Google Scholar
  13. 13.
    Davis PK, Ho A, Dowdy SF (2001) Biological methods for cell-cycle synchronization of mammalian cells. BioTechniques 30:1322–1331PubMedGoogle Scholar
  14. 14.
    Shenker JB, Datar S, Mansfield K et al (1997) Induction of apoptosis in human T-cells by organomercuric compounds: a flow cytometric analysis. Toxicol Appl Pharmacol 143:397–406CrossRefPubMedGoogle Scholar
  15. 15.
    Yoo CB, Han KT, Cho KS, Ha J, Park HJ et al (2005) Eugenol isolated from the essential oil of Eugenia caryophyllata induces a reactive oxygen species-mediated apoptosis in HL-60 human promyelocytic leukemia cells. Cancer Lett 225:41–52CrossRefPubMedGoogle Scholar
  16. 16.
    Crowley CL, Payne CM, Bernstein H (2000) The NAD+ precursors, nicotinic acid and nicotinamide protect cells against apoptosis induced by a multiple stress inducer, deoxycholate. Cell Death Differ 7:314–326CrossRefPubMedGoogle Scholar
  17. 17.
    Bestwick S, Miln L (2005) Influence of galangin on HL-60 cell proliferation and survival. Cancer Lett 243:80–89CrossRefGoogle Scholar
  18. 18.
    Jaganathan SK, Mahitosh M (2009) Antiproliferative effects of honey and of its polyphenols A review. J Biomed Biotechnol; in pressGoogle Scholar
  19. 19.
    Jaganathan SK, Mandal SM, Jana SK, Das S, Mandal M (2009) Studies on the phenolic profiling, anti-oxidant and cytotoxic activity of Indian honey: in-vitro Evaluation. Nat Prod Res; in pressGoogle Scholar
  20. 20.
    Lee YJ, Kuo HC, Chu CY, Wang CJ, Lin WC, Tseng TH (2003) Involvement of tumor suppressor protein p53 and p38 MAPK in caffeic acid phenethyl ester-induced apoptosis of C6 glioma cells. Biochem Pharmacol 66:2281–2289CrossRefPubMedGoogle Scholar
  21. 21.
    Buttke TM, Sandstrom PA (1994) Oxidative stress as a mediator of apoptosis. Immunol Today 15:7–10CrossRefPubMedGoogle Scholar
  22. 22.
    Hampton MB, Orrenius S (1998) Redox regulation of apoptotic cell death. Biofactors 8:1–5CrossRefPubMedGoogle Scholar
  23. 23.
    Davis W, Ronai Z, Tew KD (2001) Cellular thiols and reactive oxygen species in drug-induced apoptosis. J Pharmacol Exp Ther 296:1–6PubMedGoogle Scholar
  24. 24.
    Yang CF, Shen HM, Ong CN (2000) Ebselen induces apoptosis in HepG2 cells through rapid depletion of intracellular thiols. Arch Biochem Biophys 374:142–152CrossRefPubMedGoogle Scholar
  25. 25.
    Costantini P, Belzacq AS, Vieira HL et al (2000) Oxidation of a critical thiol residue of the adenine nucleotide translocator enforces Bcl-2- independent permeability transition pore opening and apoptosis. Oncogene 19:307–314CrossRefPubMedGoogle Scholar
  26. 26.
    Constantini PC, Chernyak BC, Petronilli V et al (1996) Modulation of the mitochondrial permeability transition pore by pyridine nucleotides and dithiol oxidation at two separate sites. J Biochem Biol 271:6746–51Google Scholar
  27. 27.
    Siyuan Z, Choon NO, Han MS (2004) Critical roles of intracellular thiols and calcium in parthenolide-induced apoptosis in human colorectal cancer cells. Cancer Lett 208:143–153CrossRefGoogle Scholar
  28. 28.
    Jin L, Han MS, Choon NO (2001) Role of intracellular thiol depletion, mitochondrial dysfunction and reactive oxygen species in Salvia Miltiorrhiza-induced apoptosis in human hepatoma HepG2 cells. Life Sci 69:1833–1850CrossRefGoogle Scholar
  29. 29.
    Yoko Y, Yukiko K, Rama DS et al (2002) Aluminum toxicity is associated with mitochondrial dysfunction and the production of reactive oxygen species in plant cells. Plant Physiol 128:63–72CrossRefGoogle Scholar
  30. 30.
    Simbula G, Columbano A, Ledda CG et al (2007) Increased ROS generation and p53 activation in α-lipoic acid-induced apoptosis of hepatoma cells. Apoptosis 12:113–123CrossRefPubMedGoogle Scholar
  31. 31.
    Salvador M, Makoto I, Petra B et al (2003) Influence of induced reactive oxygen species in p53-mediated cell fate decisions. Mol Cell Biol 23:8576–8585CrossRefGoogle Scholar
  32. 32.
    Kyung JW, Yong JJ, Jong WP et al (2004) Chrysin-induced apoptosis is mediated through caspase activation and Akt inactivation in U937 leukemia cells. Biochem Biophys Res Commun 325:1215–1222CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.School of Medical Science and TechnologyIndian Institute of TechnologyKharagpurIndia

Personalised recommendations