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DOI: 10.1016/j.homp.2017.09.001
Therapeutic potential of HIV nosode 30c as evaluated in A549 lung cancer cells
Subject Editor:
Publication History
Received21 April 2017
revised11 September 2017
accepted11 September 2017
Publication Date:
02 January 2018 (online)
Objectives: To examine if HIV nosode in 30c dilution (HIV 30c) has therapeutic potential against lung cancer cells (A549) as compared to WRL-68 normal cells and to elucidate its possible molecular mechanism of action on DNA replication and apoptosis.
Methods: Effects of HIV 30c were thoroughly tested for its possible anticancer potential on A549 cells (lung cancer); WRL-68 normal liver cells served as control. Three doses, one at LD50 and two below LD-50, were used. Proliferation, migration and senescence assays were made and generation of reactive oxygen species (ROS) studied by routine techniques. The ability of HIV 30c to induce apoptosis in A549 cells and its possible signalling pathway were determined using immunoblots of relevant signal proteins and confocal microscopy, including studies on telomerase reverse transcriptase (TERT) and topoisomerase II (Top II) activities, intimately associated with cell division and DNA replication.
Results: HIV 30c prevented cancer cell proliferation and migration, induced pre-mature senescence, enhanced pro-apoptotic signal proteins like p53, bax, cytochrome c, caspase-3 and inhibited anti-apoptotic signal proteins Bcl2, TERT and Top II, changed mitochondrial membrane potential and caused externalization of phosphatidyl serine. Thus, it induced apoptosis as also evidenced from increase in cells with distorted membrane morphology, nuclear condensation, DNA fragmentation, and ROS, typical of apoptosis in progress.
Conclusion: HIV 30c nosode has therapeutic potential for inducing cytotoxic effects on A549 cells as manifested by changes in nuclear condensation, DNA fragmentation, ROS generation and MMP, and for its inhibitory action on cell proliferation, cell migration, expression of telomerase reverse transcriptase and Top II genes, and increasing expression of pro-apoptotic genes.
Graphical abstract
Highlights
• HIV nosode in 30th centesimal dilution induces apoptosis in cancer cells.
• It acts by inhibiting cell proliferation and migration in A549 cells.
• It down-regulates expression of TERT and topoisomerase II enzymes.
• It induces conformational changes in DNA with associated senescence.
Keywords
HIV nosode - Anticancer potential - A549 cells - Telomerase reverse transcriptase - Topoisomerase-IIa Formerly at: Department of Zoology, University of Kalyani, 741235, India.
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References
- 1 Sepkowitz K.A. AIDS—the first 20 years. N Engl J Med 2004; 344: 1764-1772. http://www.nejm.org/doi/full/10.1056/NEJM200106073442306#t=article.
- 2 Kirch W. Encyclopedia of public health. New York: Springer; 2008: 676-677. https://books.google.co.in/books?id=eSPK7-CHw7oC&pg=PA676&redir_esc=y#v=onepage&q&f=false.
- 3 Barré-Sinoussi F., Chermann J.C., Rey F. et al. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 1983: 868-871 [Pubmed] https://www.ncbi.nlm.nih.gov/pubmed/6189183.
- 4 Conte J.E., Hadley W.K., Sande M. Infection-control guidelines for patients with the acquired immunodeficiency syndrome (AIDS). N Engl J Med 1983; 309: 740-744.
- 5 Littlewood R.A., Vanable P.A. Complementary and alternative medicine use among HIV-positive people: research synthesis and implications for HIV care. AIDS Care 2008; 20: 1002-1018 [Pubmed] https://www.ncbi.nlm.nih.gov/pubmed/18608078.
- 6 Lutge E.E., Gray A., Siegfried N. The medical use of cannabis for reducing morbidity and mortality in patients with HIV/AIDS. Cochrane Database Syst Rev 2013: 30 CD005175. [Pubmed] https://www.ncbi.nlm.nih.gov/pubmed/23633327.
- 7 Ullman D. Discovering homeopathy: medicines for 21st century. Revised edn. Berkeley, California 94712: North Atlantic books; 1991: 258.
- 8 Rastogi D.P. Asymptomatic HIV carriers. Br Homoeopath J 1994; 83: 54
- 9 Rastogi D.P., Singh V.P., Singh V. et al. Homeopathy in HIV infection: a trial report of double-blind placebo controlled study. Br Homoeopath J 1999; 88: 49-57 [Pubmed] https://www.ncbi.nlm.nih.gov/pubmed/10335412.
- 10 Singh L.M., Gupta G. Antiviral efficacy of homoeopathic drugs against animal viruses. Br Homoeopath J 1985; 74: 168-174.
- 11 Allen H.C. Materia Medica of nosodes. New Delhi: B. Jain Publ.; 1988: 139.
- 12 Mondal J., Samadder A., Khuda-Bukhsh A.R. Psorinum 6× triggers apoptosis signals in human lung cancer cells. J Integr Med 2016; 14: 143-153. http://www.jcimjournal.com/jim/showReferPage.aspx?articleTitle=Psorinum+6%C3%97+triggers+apoptosis+signals+in+human+lung+cancer+cells.
- 13 Shah R.H.I.V. nosode:The human pathagenetic trial. Forsch Komplementmed 2015; 22: 156-162 [Pubmed] https://www.ncbi.nlm.nih.gov/pubmed/26335635?dopt=Abstrac.
- 14 Hahnemann S. Organon of medicine. 6th edn. New Delhi: B. Jain Publ; 1979: 190-212.
- 15 Central Council for Research in Homeopathy: general directions and instructions for proving of drugs. http://ccrhindia.org/drugprovingintro.asp. Accessed 11 June 2015.
- 16 European Committee for Homeopathy: homeopathic drug proving guidelines. www.homeopathyeurope.org/publications/guidelines/homeopathicprovings/ECH_Proving_Guidelines-vl.pdf. Accessed 11 June 2015.
- 17 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.
- 18 Matassov D., Kagan T., Leblanc J. et al. Measurement of apoptosis by DNA fragmentation. Methods Mol Biol 2004; 282: 1-17.
- 19 Bhattacharyya S.S., Paul S., De A. et al. Poly (lactide-co-glycolide) acid nanoencapsulation of synthetic coumarin: cytotoxicity and bio-distribution in mice, in cancer cell line and interaction with calf thymus DNA as target. Toxicol Appl Pharmacol 2011; 253: 270-281.
- 20 Hulkower K.I., Herber R.L. Cell migration and invasion assays as tools for drug discovery. Pharmaceutics 2011; 3: 107-124.
- 21 Itahana K., Campisi J., Dimri G.P. Methods to detect biomarkers of cellular senescence: the senescence-associated beta-galactosidase assay. Methods Mol Biol 2007; 371: 21-31.
- 22 van Engeland M., Ramaekers F.C., Schutte B. et al. A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture. Cytometry 1996; 24: 131-139.
- 23 Klein C.A. Cancer. The metastasis cascade. Science 2008; 321: 1785-1787.
- 24 Giaccone G. DNA topoisomerases and topoisomerase inhibitors. Pathol Biol Paris 1994; 42: 346-352.
- 25 Wang J.C. Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 2002; 3: 430-440.
- 26 Willmore E., de Caux S., Sunter N.J. et al. A novel DNA-dependent protein kinase inhibitor, NU7026, potentiates the cytotoxicity of topoisomerase II poisons used in the treatment of leukemia. Blood 2004; 103: 4659-4665.
- 27 Jackson S.P. Sensing and repairing DNA double-strand breaks. Carcinogenesis 2002; 23: 687-696.
- 28 Abbotts R., Thompson N., Madhusudan S. DNA repair in cancer: emerging targets for personalized therapy. Cancer Manag Res 2014; 6: 77-92.
- 29 Roninson I.B. Tumor cell senescence in cancer treatment. Cancer Res 2003; 63: 2705-2715.
- 30 Kim N.W., Piatyszek M.A., Prowse K.R. et al. Specific association of human telomerase activity with immortal cells and cancer. Science 1994; 266: 2011-2015.
- 31 Kyo S., Inoue M. Complex regulatory mechanisms of telomerase activity in normal and cancer cells: how can we apply them for cancer therapy?. Oncogene 2002; 21: 688-697.
- 32 Shay J.W., Zou Y., Hiyama E. et al. Telomerase and cancer. Hum Mol Genet 2001; 10: 677-685.
- 33 Liu Y., Jiao R., Ma Z. et al. Sanguinarine inhibits angiotensin II-induced apoptosis in H9c2 cardiac cells via restoring reactive oxygen species-mediated decreases in the mitochondrial membrane potential. Mol Med Rep 2015; 12: 3400-3408.
- 34 Khuda-Bukhsh A.R. Potentized homeopathic drugs act through regulation of gene-expression: a hypothesis to explain their mechanism and pathways of action in vivo. Complement Ther Med 1997; 5: 43-46.
- 35 Khuda-Bukhsh A.R. Towards understanding molecular mechanisms of action of homeopathic drugs: an overview. Mol Cell Biochem 2003; 253: 339-345.
- 36 Mallick P., Mallick J.C., Guha B., Khuda-Bukhsh A.R. Ameliorating effect of microdoses of a potentized homeopathic drug, Arsenicum Album, on arsenic-induced toxicity in mice. BMC Complement Altern Med 2003; 3: 7 Oct 22.
- 37 Khuda-Bukhsh A.R. Laboratory research in homeopathy. Pro Integr Cancer Ther 2006; 5: 320-332.
- 38 Khuda-Bukhsh A.R. Mice as a model for homeopathy research. Homeopathy 2009; 98: 267-279.
- 39 Khuda-Bukhsh A.R. Current trends in ultra-high dilution research with particular reference to gene regulatory hypothesis: review. The Nucleus 2014; 57 (01) 3-17. 10.1007/s13237-014-0105-0.
- 40 Khuda-Bukhsh A.R. An overview of research at University of Kalyani in exploring some basic issues of Homoeopathy. Indian J Res Homoeopathy 2017; 11: 147-157.
- 41 Bellavite P., Signorini A., Marzotto M. et al. Cell sensitivity, non-linearity and inverse effects. Homeopathy 2015; 104: 139-160.
- 42 Marzotto M., Bonafini C., Olioso D. et al. Arnica montana stimulates extracellular matrix gene expression in a macrophage cell line differentiated to wound-healing phenotype. PLoS One 2016; 11: e0166340.