Static Magnetic Field Reduces the Anticancer Effect of Hinokitiol on Melanoma Malignant Cells-Gene Expression and Redox Homeostasis Studies.

BACKGROUND: Melanoma malignant is characterized by a high mortality rate, accounting for as much as 65% of deaths caused by skin cancer. A potential strategy in cancer treatment may be the use of natural compounds, which include hinokitiol (ß-Thujaplicin), a phenolic component of essential oils extracted from cypress trees. Many studies confirm that a high-induction SMF (static magnetic field) has anticancer effects and can be used as a non-invasive anticancer therapy in combination with or without drugs. AIM: The aim of this experiment was to evaluate the effect of a static magnetic field on melanoma cell cultures (C32 and COLO 829) treated with hinokitiol. METHODS AND RESULTS: Melanoma cells were exposed to a static magnetic field of moderate induction and hinokitiol. The research included determining the activity of the antioxidant enzymes (SOD, GPx, and CAT) and MDA concentration as well as the gene expression profile. CONCLUSION: Hinokitiol disturbs the redox homeostasis of C32 and COLO 829 melanoma malignant cells. Moreover, a static magnetic field has a protective effect on melanoma malignant cells and abolishes the anticancer effect of hinokitiol.

Phenolic acids and a static magnetic field change the expression of transforming growth factor ß isoforms in amelanotic melanoma cells.

BACKGROUND: Melanoma is an aggressive type of cancer that can metastasize to numerous other organs. TGFß is one of the key signaling pathways in melanoma progression. Previous studies on various types of cancer have shown that both: polyphenols and a static magnetic field (SMF) can be potential chemopreventive/therapeutic agents. Therefore, the aim of the study was to evaluate the effect of a SMF and selected polyphenols on the transcriptional activity of TGFß genes in melanoma cells. METHODS AND RESULTS: Experiments were performed on the C32 cell line treated with caffeic or chlorogenic acids, and with simultaneous exposure to a moderate-strength SMF. The RT-qPCR method was used to determine the mRNA level of genes encoding the TGFß isoforms and their receptors. The concentration of the TGFß1 and TGFß2 proteins were also measured in the cell culture supernates. The first response of C32 melanoma cells to both factors is the reduction of TGFß levels. Then, mRNA level of these molecules returned to values close to pre-treatment level by the end of experiment. CONCLUSION: Our study results demonstrate the potential of polyphenols and a moderate-strength SMF to support cancer therapy by altering TGFß expression, which is a very promising topic for the diagnosis and treatment of melanoma.

The Apoptotic Effect of Caffeic or Chlorogenic Acid on the C32 Cells That Have Simultaneously Been Exposed to a Static Magnetic Field.

The induction of apoptosis is one of the main goals of the designed anti-cancer therapies. In recent years, increased attention has been paid to the physical factors such as magnetic fields and to the natural bioactive compounds and the possibilities using them in medicine. Hence, the aim of this study was to evaluate the anti-tumor effect of caffeic or chlorogenic acid in combination with a moderate-strength static magnetic field on C32 melanoma cells by assessing the effect of both factors on the apoptotic process. The apoptosis of the C32 cells was evaluated using a flow cytometry analysis. The expression of the apoptosis-associated genes was determined using the RT-qPCR technique. The caspase activity and the concentration of the oxidative damage markers were also measured. It was found that phenolic acids and a static magnetic field trigger the apoptosis of the C32 cells and also affect the expression of the genes encoding the apoptosis regulatory proteins. In conclusion, our study indicated that both of the phenolic acids and a static magnetic field can be used supportively in the treatment of melanoma and that caffeic acid is more pro-apoptotic than chlorogenic acid.

The Influence of Betulin and Its Derivatives EB5 and ECH147 on the Antioxidant Status of Human Renal Proximal Tubule Epithelial Cells.

Betulin and its derivatives, 28-propyne derivative EB5 and 29-diethyl phosphonate analog ECH147, are promising compounds in anti-tumor activity studies. However, their effect on kidney cells has not yet been studied. The study aimed to determine whether betulin and its derivatives-EB5 and ECH147-influence the viability and oxidative status of human renal proximal tubule epithelial cells (RPTECs). The total antioxidant capacity of cells (TEAC), lipid peroxidation product malondialdehyde (MDA) level, and activity of antioxidant enzymes (SOD, CAT, and GPX) were evaluated. Additionally, the mRNA level of genes encoding antioxidant enzymes was assessed. Cisplatin and 5-fluorouracil were used as reference substances. Betulin and its derivatives affected the viability and antioxidant systems of RPTECs. Betulin strongly reduced TEAC in a concentration-dependent manner. All tested compounds caused an increase in MDA levels. The activity of SOD, CAT, and GPX, and the mRNA profiles of genes encoding antioxidant enzymes depended on the tested compound and its concentration. Betulin showed an cisplatin-like effect, indicating its nephrotoxic potential. Betulin derivatives EB5 and ECH147 showed different impacts on the antioxidant system, which gives hope that these compounds will not cause severe consequences for the kidneys in vivo.

The effect of a static magnetic field and baicalin or baicalein interactions on amelanotic melanoma cell cultures (C32).

BACKGROUND: Baicalin and baicalein have antioxidant, anti-inflammatory, hepatoprotective and anti-cancer properties. However, it is not known how a static magnetic field will modify these properties. Therefore, the aim of our study was to evaluate the simultaneous exposure of melanoma cells to flavones and the static magnetic fields that are generated by permanent magnets on the gene expression and the activity of the antioxidant enzymes that are associated with the antioxidant defense system. METHODS AND RESULTS: Melanoma cells that had been treated with baicalin or baicalein were subjected to a static magnetic fields with a moderate induction. The static magnetic field was emitted by permanent magnets and the cell cultures were carried out in special test chambers. The research included determining the activity of the antioxidant enzymes (superoxide dismutase, glutathione peroxidase and catalase) as well as the gene expression profile. The addition of the flavones to the cell cultures at a concentration of 50 µmol/L resulted increase in the expression of the SOD1, SOD2 and GPX1 genes compared to the nontreated cell cultures. Simultaneous exposure of the melanoma cells to static magnetic field and baicalin or baicalein reduced their mRNA levels compared to the cultures to which only baicalin or baicalein had been added. The change in gene expression was accompanied by changes at the protein level associated with an increase in the activity of antioxidant enzymes. CONCLUSION: We showed that baicalin or baicalein have anticancer properties by disturbing the redox homeostasis in melanoma cells and also increases the antioxidant system gene expression. There was also an antagonistic interaction between the studied flavones and the static magnetic field, which cause a decrease in the anticancer effects of baicalin or baicalein.

The impact of the co-exposure of melanoma cells to chlorogenic acid and a moderate-strength static magnetic field.

A static magnetic field (SMF) or the bioactive compounds that are found in foods are potential agents that can be used to support cancer therapy. Therefore, the aim of our study was to assess the impact of the SMF that are induced by neodymium magnets on the culture growth and antioxidant status of melanoma cells that had been treated with chlorogenic acid (CGA). The melanoma cells, the control and those that had been treated with CGA, were put in special magnetic test chambers that generated a 0.7 T magnetic field. The mRNA levels of the antioxidant enzymes were analyzed using RT-qPCR. The activity of SOD, GPx, and CAT was measured in the cell lysates. While the expression and activity of the antioxidant enzymes was inhibited relative to the untreated cells as a result of the CGA treatment (1 mmol/L), it was not after the CGA treatment in combination with an SMF. The demonstrated cytotoxicity of CGA (1 mmol/L) and its inhibition of the antioxidant enzymes suggests the usefulness of phenolic compounds as a supporting pharmacological treatment for melanoma. PRACTICAL APPLICATIONS: Phenolic acids and their derivatives, which are the bioactive components of the human diet, are signal molecules that transfer information from the external environment that affects the level of gene expression in cells. This study suggests the usefulness of phenolic compounds as a supporting pharmacological treatment for melanoma and seems to be important for the development of experimental oncology.

Impact of fluoride and a static magnetic field on the gene expression that is associated with the antioxidant defense system of human fibroblasts.

Fluoride cytotoxicity has been associated with apoptosis, oxidative stress, general changes in DNA and RNA and protein biosynthesis, whereas the results of studies on the effect of SMF on antioxidant activity of cells are contradictory. Therefore, the aim of our study was to evaluate the simultaneous exposure of human cells to fluoride SMF that are generated by permanent magnets on the expression profile of the genes that are associated with the antioxidant defense system. Control fibroblasts and fibroblasts that had been treated with fluoride were subjected to the influence of SMF with a moderate induction. In order to achieve our aims, we applied modern molecular biology techniques such as the oligonucleotide microarray. Among the antioxidant defense genes, five (SOD1, PLK3, CLN8, XPA, HAO1), whose expression was significantly altered by the action of fluoride ions and the exposure to SMF were normalized their expression was identified. We showed that fluoride ions cause oxidative stress, whereas exposure to SMF with a moderate induction can suppress their effects by normalizing the expression of the genes that are altered by fluoride. Our research may explain the molecular mechanisms of the influence of fluoride and SMF that are generated by permanent magnets on cells.

Influence of static magnetic fields up to 700 mT and dihydrochalcones on the antioxidant response in fibroblasts.

The effects of a static magnetic field (SMF) and the dihydrochalcones phloretin and phloridzin on the redox homeostasis of fibroblasts were investigated. The aim of the present study was to determine the redox homeostasis of fibroblasts that were simultaneously exposed to a static magnetic field and the dihydrochalcones phloretin and phloridzin. The fibroblasts were cultured for 72 h in special magnetic test chambers at different moderate intensities (0.4, 0.55 and 0.7 T). In this report, the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione transferase (GST); the concentrations of malondialdehyde (MDA), adenosine triphosphate (ATP) and total antioxidant status were measured using commercially available kits. We did not observe any impairment in the redox balance in cells in fibroblasts that were only exposed to static magnetic fields of different intensities or In fibroblast cultured with dihydrochalcones and exposed to static magnetic field increase the SOD, GPx, GST activities and MDA concentration. Our investigations revealed that the activities of SOD, GPx, GST and the concentration of MDA that were determined for the fibroblasts that were cultured with dihydrochalcones were higher in the presence of a static magnetic field. Our results indicated that exposure to SMF (0.7 T) with dihydrochalcones induces oxidative stress in fibroblasts.

Studies on the effect of quercetin and nitrates on the redox homeostasis using in vitro model.

Antioxidants are widely considered to be a preventive measure for many diseases and beneficial for health. However, an increasing number of reports suggest a lack of any influence by antioxidants on health or even harmful pro-oxidative effects of antioxidants. In most cases, the research was conducted with respect to a chosen antioxidant, without considering the presence of other chemical substances present in food, with which these compounds may react. The aim of this work was to determine whether and to what extent the simultaneous presence of quercetin and sodium nitrate influences oxidative-reductive homeostasis in fibroblast cultures. Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and nitric oxide synthase (NOS) activities were measured together with nitric oxide (NO) concentration and total antioxidant status (TAS). An increase in the activity of all the enzymes measured and in the NO concentration was determined compared with the control culture. The most prominent changes were observed at the highest quercetin concentration. These results indicate that the simultaneous presence of quercetin and sodium nitrate disrupts the oxidative-reductive homeostasis in fibroblasts.

Effect of a static magnetic fields and fluoride ions on the antioxidant defense system of mice fibroblasts.

The results of studies on the biological influence of magnetic fields are controversial and do not provide clear answers regarding their impact on cell functioning. Fluoride compounds are substances that influence free radical processes, which occur when the reactive forms of oxygen are present. It is not known whether static magnetic fields (SMF) cause any changes in fluoride assimilation or activity. Therefore, the aim of this work was to determine the potential relationship between magnetic field exposure to, and the antioxidant system of, fibroblasts cultured with fluoride ions. Three chambers with static magnetic fields of different intensities (0.4, 0.6, and 0.7 T) were used in this work. Fluoride ions were added at a concentration of 0.12 mM, which did not cause the precipitation of calcium or magnesium. The results of this study show that static magnetic fields reduce the oxidative stress caused by fluoride ions and normalize the activities of antioxidant enzymes, including superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). Static magnetic fields modify the energy state of fibroblasts, causing an increase in the ATP concentration and a decrease in the MDA concentration. These results suggest that exposure to fluoride and an SMF improves the tolerance of cells to the oxidative stress induced by fluoride ions.