Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
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 p21WAF, 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 p21WAF, 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.
Abstract
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 p21WAF, 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 p21WAF, 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.
참고문헌 (35)
- Ernst E, "A systematic review of systematic reviews of homeopathy", Br J Clin Pharmacol, 54(6), pp.577~582 (2002)
- Shang A, "Are the clinical effects of homoeopathy placebo effects? Comparative study of placebo-controlled trials of homoeopathy and allopathy", Lancet, 366(9487), pp.726~732 (2005)
- Clausen J, "Review of the use of high potencies in basic research on homeopathy", Homeopathy, 100(4), pp.288~292 (2011)
- Bellavite P, "Advances in homeopathy and immunology: a review of clinical research", Front Biosci, 3, pp.1363~1389 (2011)
- Bornhoft G, "Homeopathy in healthcare effectiveness, appropriateness, safety, cost", pp.209~ (2011), Spinger-Verlag
- Demangeat JL., "NMR relaxation evidence for solute-induced nanosized superstructures in ultramolecular aqueous dilutions of silica lactose", J Molecular Liquids, 155(2-3), pp.71~79 (2010)
- Liu L, "Particles shed from syringe filters and their effects on agitation-induced protein aggregation", J Pharm Sci, 101(8), pp.2952~2959 (2012)
- Chikramane PS, "Extreme homeopathic dilutions retain starting materials: a nanoparticulate perspective", Homeopathy, 99(4), pp.231~242 (2010)
- Bell IR, "Testing the nanoparticle- allostatic cross-adaptation-sensitization model for homeopathic remedy effects", Homeopathy, 102(1), pp.66~81 (2013)
- Bhattacharyya SS, "Encapsulated plant extract (Gelsemium sempervirens) poly (lactide-co-glycolide) nanoparticles enhance cellular uptake and increase bioactivity in vitro", Exp Biol Med, 235(6), pp.678~688 (2010)
- Chikramane PS, "Why extreme dilutions reach non-zero asymptotes: a nanoparticulate hypothesis based on froth flotation", Langmuir, 28(45), pp.15864~15875 (2012)
- Bel Haaj S, "Starch nanoparticles formation via high-power ultrasonication", Carbohydr Polym, 92(2), pp.1625~1632 (2013)
- Khuda-Bukhsh AR, "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, pp.2011~ (2011)
- 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, 5(1), pp.43~46 (1997)
- Khuda-Bukhsh AR, "Towards understanding molecular mechanisms of action of homeopathic drugs: an overview", Mol Cell Biochem, 253(1-2), pp.339~345 (2003)
- Khuda-Bukhsh AR, "Laboratory research in homeopathy: pro", Integr Cancer Ther, 5(4), pp.320~332 (2006)
- Khuda-Bukhsh AR, "Homeopathic drug discovery: theory update and methodological aspect", Expert Opin Drug Discov, 3(8), pp.979~990 (2008)
- Mei S, "Role of histone deacetylase inhibitors in the treatment of cancer (Review)", Int J Oncol, 25(6), pp.1509~1519 (2004)
- Bishayee K, "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, 382(1-2), pp.173~183 (2013)
- Ghosh S, "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, 11(2), pp.116~124 (2013)
- Mukherjee A, "Ethanolic extract of Thuja occidentalis blocks proliferation of A549 cells and induces apoptosis in vitro", Zhong Xi Yi Jie He Xue Bao, 10(12), pp.1451~1459 (2012)
- Bishayee K, "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, 2013, pp.46(2), 153~163
- Kurdistani SK, "Mapping global histone acetylation patterns to gene expression", Cell, 117(6), pp.721~733 (2004)
- Bolden JE, "Anticancer activities of histone deacetylase inhibitors", Nat Rev Drug Discov, 5(9), pp.769~784 (2006)
- Kelly TK, "Epigenetic modification as therapeutic targets", Nat Biotechnol, 28(10), pp.1069~1078 (2010)
- Ward CS, "HDAC inhibition induces increased choline uptake and elevated phosphocholine levels in MCF7 breast cancer cells", PLoS One, 8(4), pp.e62610~ (2013)
- Hrzenjak A, "Valproate inhibition of histone deacetylase 2 affects differentiation and decreases proliferation of endometrial stromal sarcoma cells", Mol Cancer Ther, 5(9), pp.2203~2210 (2006)
- Warrell RP Jr, "Therapeutic targeting of transcription in acute promyelocytic leukemia by use of an inhibitor of histone deacetylase", J Natl Cancer Inst, 90(21), pp.1621~1625 (1998)
- Dokmanovic M, "Histone deacetylase inhibitors: overview and perspectives", Mol Cancer Res, 5(10), pp.981~989 (2007)
- Sterner DE, "Acetylation of histones and transcription-related factors", Microbiol Mol Biol Rev, 64(2), pp.435~439 (2000)
- Ropero S, "The role of histone deacetylases (HDACs) in human cancer", Mol Oncol, 1(1), pp.19~25 (2007)
- Vigushin DM, "Histone deacetylase inhibitors in cancer treatment", Anticancer Drugs, 13(1), pp.1~13 (2002)
- Icardi L, "Opposed regulation of type I IFN-induced STAT3 and ISGF3 transcriptional activities by histone deacetylases (HDACS) 1 and 2", FASEB J, 26(1), pp.240~249 (2012)
- Condorelli G, "Mutated p21", WAF, FASEB J, pp.2001, 15(2)[DOI]
- Macleod KF, "p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage", Genes Dev, 9(8), pp.935~944 (1995)