대한약침학회지 (Journal of Pharmacopuncture)

  • 제16권4호
  • /
  • Pages.7-13
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  • 2013
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  • 2093-6966(pISSN)
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  • 2234-6856(eISSN)
대한약침학회 (KOREAN PHARMACOPUNCTURE INSTITUTE)
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Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract

  • Bishayee, Kausik (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani) ;
  • Sikdar, Sourav (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani) ;
  • Khuda-Bukhsh, Anisur Rahman (Department of Zoology, University of Kalyani)
  • 투고 : 2013.10.17
  • 심사 : 2013.10.30
  • 발행 : 2013.12.31
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초록

Objectives: Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored. Therefore, in this study, we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C (ethanolic extract of Gonolobus condurango diluted $10^{-60}$ times) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol (placebo) control. Methods: We checked the activity of different signal proteins (like $p21^{WAF}$, p53, Akt, STAT3) related to deacetylation, cell growth and differentiation by western blotting and analyzed cell-cycle arrest, if any, by fluorescence activated cell sorting. After viability assays had been performed with Condurango 30C and with a placebo, the activities of histone de-acetylase (HDAC) enzymes 1 and 2 were measured colorimetrically. Results: While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced HDAC2 activity quite strikingly, it apparently did not alter the HDAC1 enzyme; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity. Data on $p21^{WAF}$, p53, Akt, and STAT3 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and G1-phase cell-cycle arrest when Condurango 30C was used at a 2% dose. Conclusion: Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells, which the placebo was unable to do.

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참고문헌

  1. Ernst E. A systematic review of systematic reviews of homeopathy. Br J Clin Pharmacol. 2002;54(6):577-82. https://doi.org/10.1046/j.1365-2125.2002.01699.x
  2. Shang A, Huwiler-Muntener K, Nartey L, Jüni P, Dörig S, Sterne JA, et al. Are the clinical effects of homoeopathy placebo effects? Comparative study of placebo- controlled trials of homoeopathy and allopathy Lancet. 2005;366(9487):726-32.
  3. Clausen J, van Wijk R, Albrecht H. Review of the use of high potencies in basic research on homeopathy. Homeopathy. 2011;100(4):288-92. https://doi.org/10.1016/j.homp.2011.07.003
  4. Bellavite P, Marzotto M, Chirumbolo S, Conforti A. Advances in homeopathy and immunology: a review of clinical research. Front Biosci (Schol Ed). 2011;3:1363-89.
  5. Bornhoft G, Matthiessen PF. Homeopathy in healthcare effectiveness, appropriateness, safety, cost. Berlin: Spinger-Verlag; 2011. 209 p.
  6. Demangeat JL. NMR relaxation evidence for solute-induced nanosized superstructures in ultramolecular aqueous dilutions of silica lactose. J Molecular Liquids. 2010;155(2-3):71-9. https://doi.org/10.1016/j.molliq.2010.05.010
  7. Liu L, Randolph TW, Carpenter JF. Particles shed from syringe filters and their effects on agitation-induced protein aggregation. J Pharm Sci. 2012;101(8):2952-9. https://doi.org/10.1002/jps.23225
  8. Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homeopathic dilutions retain starting materials: a nanoparticulate perspective. Homeopathy. 2010;99(4):231-42. https://doi.org/10.1016/j.homp.2010.05.006
  9. Bell IR, Koithan M, Brooks AJ. Testing the nanoparticle- allostatic cross-adaptation-sensitization model for homeopathic remedy effects. Homeopathy. 2013;102(1):66-81. https://doi.org/10.1016/j.homp.2012.10.005
  10. Bhattacharyya SS, Paul S, Khuda-Bukhsh AR. Encapsulated plant extract (Gelsemium sempervirens) poly (lactide-co-glycolide) nanoparticles enhance cellular uptake and increase bioactivity in vitro. Exp Biol Med (Maywood). 2010;235(6):678-88. https://doi.org/10.1258/ebm.2010.009338
  11. Chikramane PS, Kalita D, Suresh AK, Kane SG, Bellare JR. Why extreme dilutions reach non-zero asymptotes: a nanoparticulate hypothesis based on froth flotation. Langmuir. 2012;28(45):15864-75. https://doi.org/10.1021/la303477s
  12. Bel Haaj S, Magnin A, Pétrier C, Boufi S. Starch nanoparticles formation via high-power ultrasonication. Carbohydr Polym. 2013;92(2):1625-32. https://doi.org/10.1016/j.carbpol.2012.11.022
  13. Khuda-Bukhsh AR, Bhattacharyya SS, Paul S, Dutta S, Boujedaini N, Belon P. Modulation of signal proteins: a plausible mechanism to explain how a potentized drug secale cor 30C diluted beyond Avogadro's limit combats skin papilloma in mice. Evid Based Complement Alternat Med. 2011;2011.
  14. Khuda-Bukhsh AR. Potentized homoeopathic drugs act through regulation of gene-expression: a hypothesis to explain their mechanism and pathways of action in vitro. Complement Ther Med. 1997;5(1):43-6. https://doi.org/10.1016/S0965-2299(97)80090-8
  15. Khuda-Bukhsh AR. Towards understanding molecular mechanisms of action of homeopathic drugs: an overview. Mol Cell Biochem. 2003;253(1-2):339-45. https://doi.org/10.1023/A:1026048907739
  16. Khuda-Bukhsh AR. Laboratory research in homeopathy: pro. Integr Cancer Ther. 2006;5(4):320-32. https://doi.org/10.1177/1534735406294794
  17. Khuda-Bukhsh AR, Pathak S. Homeopathic drug discovery: theory update and methodological aspect. Expert Opin Drug Discov. 2008;3(8):979-90. https://doi.org/10.1517/17460441.3.8.979
  18. Mei S, Ho AD, Mahlknecht U. Role of histone deacetylase inhibitors in the treatment of cancer (Review). Int J Oncol. 2004;25(6):1509-19.
  19. Bishayee K, Paul A, Ghosh S, Sikdar S, Mukherjee A, Biswas R, et al. Condurango glycosides A fraction of Gonolobus condurango induces DNA-damage-associated senescence and apoptosis via an ROS-dependent p53 signalling pathway in HeLa cells. Mol Cell Biochem. 2013;382(1-2):173-83. https://doi.org/10.1007/s11010-013-1732-5
  20. Ghosh S, Bishayee K, Paul A, Mukherjee A, Sikdar S, Chakraborty D, et al. Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation. J Integr Med. 2013;11(2):116-24. https://doi.org/10.3736/jintegrmed2013014
  21. Mukherjee A, Sikdar S, Bishayee K, Paul A, Ghosh S, Boujedaini N, et al. Ethanolic extract of Thuja occidentalis blocks proliferation of A549 cells and induces apoptosis in vitro. Zhong Xi Yi Jie He Xue Bao. 2012;10(12):1451-9. https://doi.org/10.3736/jcim20121218
  22. Bishayee K, Ghosh S, Mukherjee A, Sadhukhan R, Mondal J, Khuda-Bukhsh AR. Quercetin induces cytochrome- c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interaction. Cell Prolif. 2013;46(2):153-63. https://doi.org/10.1111/cpr.12017
  23. Kurdistani SK, Tavazoie S, Grunstein M. Mapping global histone acetylation patterns to gene expression. Cell. 2004;117(6):721-33. https://doi.org/10.1016/j.cell.2004.05.023
  24. Bolden JE, Peart MJ, Johnstone RW. Anticancer activities of histone deacetylase inhibitors. Nat Rev Drug Discov. 2006;5(9):769-84. https://doi.org/10.1038/nrd2133
  25. Kelly TK, de Carvalho DD, Jones PA. Epigenetic modification as therapeutic targets. Nat Biotechnol. 2010;28(10):1069-78. https://doi.org/10.1038/nbt.1678
  26. Ward CS, Eriksson P, Izquierdo-Garcia JL, Brandes AH, Ronen SM. HDAC inhibition induces increased choline uptake and elevated phosphocholine levels in MCF7 breast cancer cells. PLoS One. 2013;8(4):e62610. https://doi.org/10.1371/journal.pone.0062610
  27. Hrzenjak A, Moinfar F, Kremser ML, Strohmeier B, Staber PB, Zatloukal K, et al. Valproate inhibition of histone deacetylase 2 affects differentiation and decreases proliferation of endometrial stromal sarcoma cells. Mol Cancer Ther. 2006;5(9):2203-10. https://doi.org/10.1158/1535-7163.MCT-05-0480
  28. Warrell RP Jr, He LZ, Richon V, Calleja E, Pandolfi PP. Therapeutic targeting of transcription in acute promyelocytic leukemia by use of an inhibitor of histone deacetylase. J Natl Cancer Inst. 1998;90(21):1621-5. https://doi.org/10.1093/jnci/90.21.1621
  29. Dokmanovic M, Clarke C, Marks PA. Histone deacetylase inhibitors: overview and perspectives. Mol Cancer Res. 2007;5(10):981-9. https://doi.org/10.1158/1541-7786.MCR-07-0324
  30. Sterner DE, Berger SL. Acetylation of histones and transcription-related factors. Microbiol Mol Biol Rev. 2000; 64(2):435-9. https://doi.org/10.1128/MMBR.64.2.435-459.2000
  31. Ropero S, Esteller M. The role of histone deacetylases (HDACs) in human cancer. Mol Oncol. 2007;1(1):19-25. https://doi.org/10.1016/j.molonc.2007.01.001
  32. Vigushin DM, Coombes RC. Histone deacetylase inhibitors in cancer treatment. Anticancer Drugs. 2002;13(1):1-13. https://doi.org/10.1097/00001813-200201000-00001
  33. Icardi L, Lievens S, Mori R, Piessevaux J, De Cauwer L, De Bosscher K, et al. Opposed regulation of type I IFN-induced STAT3 and ISGF3 transcriptional activities by histone deacetylases (HDACS) 1 and 2. FASEB J. 2012;26(1):240-9. https://doi.org/10.1096/fj.11-191122
  34. Condorelli G, Aycock JK, Frati G, Napoli C. Mutated p21/WAF/CIP transgene overexpression reduces smooth muscle cell proliferation, macrophage deposition, oxidation-sensitive mechanisms, and restenosis in hypercholesterolemic apolipoprotein E knockout mice. FASEB J. 2001;15(2):2162-70. https://doi.org/10.1096/fj.01-0032com
  35. Macleod KF, Sherry N, Hannon G, Beach D, Tokino T, Kinzler K, et al. p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage. Genes Dev. 1995;9(8):935-44. https://doi.org/10.1101/gad.9.8.935

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