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1.
Journal of Integrative Medicine ; (12): 116-124, 2013.
Article in English | WPRIM | ID: wpr-308266

ABSTRACT

<p><b>OBJECTIVE</b>Preventive measures against skin melanoma like chemotherapy are useful but suffer from chronic side effects and drug resistance. Ethanolic extract of Phytolacca decandra (PD), used in homeopathy for the treatment of various ailments like chronic rheumatism, regular conjunctivitis, psoriasis, and in some skin diseases was tested for its possible anticancer potential.</p><p><b>METHODS</b>Cytotoxicity of the drug was tested by conducting 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on both normal (peripheral blood mononuclear cells) and A375 cells. Fluorescence microscopic study of 4',6-diamidino-2-phenylindole dihydrochloride-stained cells was conducted for DNA fragmentation assay, and changes in cellular morphology, if any, were also recorded. Lactate dehydrogenase activity assay was done to evaluate the percentages of apoptosis and necrosis. Reactive oxygen species (ROS) accumulation, if any, and expression study of apoptotic genes also were evaluated to pin-point the actual events of apoptosis.</p><p><b>RESULTS</b>Results showed that PD administration caused a remarkable reduction in proliferation of A375 cells, without showing much cytotoxicity on peripheral blood mononuclear cells. Generation of ROS and DNA damage, which made the cancer cells prone to apoptosis, were found to be enhanced in PD-treated cells. These results were duly supported by the analytical data on expression of different cellular and nuclear proteins, as for example, by down-regulation of Akt and Bcl-2, up-regulation of p53, Bax and caspase 3, and an increase in number of cell deaths by apoptosis in A375 cells.</p><p><b>CONCLUSION</b>Overall results demonstrate anticancer potentials of PD on A375 cells through activation of caspase-mediated signaling and ROS generation.</p>


Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Caspase 3 , Genetics , Metabolism , Cell Line, Tumor , Drugs, Chinese Herbal , Pharmacology , Gene Expression Regulation, Neoplastic , Melanoma , Drug Therapy , Genetics , Metabolism , Phytolacca , Chemistry , Phytotherapy , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Reactive Oxygen Species , Metabolism , Signal Transduction , Skin Neoplasms , Drug Therapy , Genetics , Metabolism , Up-Regulation
2.
Journal of Integrative Medicine ; (12): 397-404, 2013.
Article in English | WPRIM | ID: wpr-308228

ABSTRACT

<p><b>OBJECTIVE</b>To examine if the ultra-highly diluted homeopathic remedy Thuja 30C can ameliorate benzo(a)pyrene (BaP)-induced DNA damage, stress and viability of perfused lung cells of Swiss albino mice in vitro.</p><p><b>METHODS</b>Perfused normal lung cells from mice were cultured in 5% Roswell Park Memorial Institute medium and exposed to BaP, a potent carcinogen, at the half maximal inhibitory concentration dose (2.2 μmol/L) for 24 h. Thereafter, the intoxicated cells were either treated with Thuja 30C (used against tumor or cancer) or its vehicle media, succussed alcohol 30C. Relevant parameters of study involving reactive oxygen species (ROS) accumulation, total glutathione (GSH) content, and generations of heat shock protein (hsp)-90 were measured; the cell viability and other test parameters were measured after treatment with either Thuja 30C or its vehicle media. Circular dichroism spectroscopy was performed to examine if Thuja 30C directly interacted with calf thymus DNA as target. For ascertaining if DNA damaged by BaP could be partially repaired and restituted by the remedy, 4',6-diamidino-2-phenylindole staining was performed.</p><p><b>RESULTS</b>Thuja 30C increased cell viability of BaP-intoxicated cells significantly, as compared to drug-untreated or drug-vehicle control. A minimal dose of Thuja 30C significantly inhibited BaP-induced stress level, by down-regulating ROS and hsp-90, and increasing GSH content. Thuja 30C itself had no DNA-damaging effect, and no direct drug-DNA interaction. However, it showed quite striking ability to repair DNA damage caused by BaP.</p><p><b>CONCLUSION</b>Thuja 30C ameliorates BaP-induced toxicity, stress and DNA damage in perfused lung cells of mice and it apparently has no effect on normal lung cells.</p>


Subject(s)
Animals , Mice , Benzo(a)pyrene , Toxicity , Cell Survival , DNA Damage , Glutathione , Metabolism , HSP90 Heat-Shock Proteins , Homeopathy , Mice, Inbred Strains , Reactive Oxygen Species , Metabolism , Thuja
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