RESUMO
Uncaria tomentosa is a medicinal plant native to Peru that has been traditionally used in the treatment of various inflammatory disorders. In this study, the effectiveness of U. tomentosa as an anti-cancer agent was assessed using the growth and survival of B16-BL6 mouse melanoma cells. B16-BL6 cell cultures treated with both ethanol and phosphate-buffered saline (PBS) extracts of U. tomentosa displayed up to 80% lower levels of growth and increased apoptosis compared to vehicle controls. Treatment with ethanolic extracts of Uncaria tomentosa were much more effective than treatment with aqueous extracts. U. tomentosa was also shown to inhibit B16-BL6 cell growth in C57/bl mice in vivo. Mice injected with both the ethanolic and aqueous extracts of U. tomentosa showed a 59 ± 13% decrease in B16-BL6 tumour weight and a 40 ± 9% decrease in tumour size. Histochemical analysis of the B16-BL6 tumours showed a strong reduction in the Ki-67 cell proliferation marker in U. tomentosa-treated mice and a small, but insignificant increase in terminal transferase dUTP nick labelling (TUNEL) staining. Furthermore, U. tomentosa extracts reduced angiogenic markers and reduced the infiltration of T cells into the tumours. Collectively, the results in this study concluded that U. tomentosa has potent anti-cancer activity that significantly inhibited cancer cells in vitro and in vivo.
Assuntos
Apoptose , Unha-de-Gato/química , Melanoma Experimental/patologia , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Fosforilação/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Carga Tumoral/efeitos dos fármacosRESUMO
Exposure to specific electromagnetic field (EMF) patterns can affect a variety of biological systems. We have shown that exposure to Thomas-EMF, a low-intensity, frequency-modulated (25-6 Hz) EMF pattern, inhibited growth and altered cell signaling in malignant cells. Exposure to Thomas-EMF for 1 h/day inhibited the growth of malignant cells including B16-BL6 mouse melanoma cells, MDA-MB-231, MDA-MB-468, BT-20, and MCF-7 human breast cancer and HeLa cervical cancer cells but did not affect non-malignant cells. The Thomas-EMF-dependent changes in cell proliferation were mediated by adenosine 3',5'-cyclic monophosphate (cAMP) and extracellular-signal-regulated kinase (ERK) signaling pathways. Exposure of malignant cells to Thomas-EMF transiently changed the level of cellular cAMP and promoted ERK phosphorylation. Pharmacologic inhibitors (SQ22536) and activators (forskolin) of cAMP production both blocked the ability of Thomas-EMF to inhibit cell proliferation, and an inhibitor of the MAP kinase pathway (PD98059) was able to partially block Thomas-EMF-dependent inhibition of cell proliferation. Genetic modulation of protein kinase A (PKA) in B16-BL6 cells also altered the effect of Thomas-EMF on cell proliferation. Cells transfected with the constitutively active form of PKA (PKA-CA), which interfered with ERK phosphorylation, also interfered with the Thomas-EMF effect on cell proliferation. The non-malignant cells did not show any EMF-dependent changes in cAMP levels, ERK phosphorylation, or cell growth. These data indicate that exposure to the specific Thomas-EMF pattern can inhibit the growth of malignant cells in a manner dependent on contributions from the cAMP and MAP kinase pathways. Bioelectromagnetics. 39;217-230, 2018. © 2017 Wiley Periodicals, Inc.
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AMP Cíclico/metabolismo , Campos Eletromagnéticos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos da radiação , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos da radiação , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Humanos , Camundongos , Fosforilação/efeitos da radiação , Fatores de TempoRESUMO
Exposure to low intensity, low frequency electromagnetic fields (EMF) has effects on several biological systems. Spatiotemporal characteristics of these EMFs are critical. The effect of several complex EMF patterns on the proliferation of B16-BL6 mouse melanoma cells was tested. Exposure to one of these patterns, the Thomas-EMF, inhibited cell proliferation and promoted calcium uptake. The Thomas-EMF is coded from a digital-to-analog file comprised of 849 points, which provides power to solenoids and can be set to alter timing, intensity, and duration of variable EMF. Setting the point duration to 3 ms generated a time-varying EMF pattern which began at 25 Hz and slowed to 6 Hz over a 2.5 s repeat. Exposing B16-BL6 cells to Thomas-EMF set to 3 ms for 1 h/day inhibited cell proliferation by 40% after 5 days, while setting the point duration to 1, 2, 4, or 5 ms had no effect on cell proliferation. Similarly, exposing cells to Thomas-EMF set to 3 ms promoted a three-fold increase in calcium uptake after 1 h, while the other timings had no effect. Exposure to Thomas-EMF for as short as 15 min/day slowed cell proliferation, but exposure for 1 h/day was optimal. This corresponded to the effect on calcium uptake where uptake was detected after 15 min exposure and was maximal by 1 h of treatment. Studies show that the specific spatiotemporal character of EMF is critical in mediating their biological activities. Bioelectromagnetics. 38:165-174, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Campos Eletromagnéticos , Melanoma Experimental/patologia , Animais , Cálcio/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Magnetoterapia , Melanoma Experimental/metabolismo , Melanoma Experimental/terapia , Camundongos , Fatores de TempoRESUMO
Exposure to ionising radiation is clearly associated with an increased risk of developing some types of cancer. However, the contribution of non-targeted effects to cancer development after exposure to ionising radiation is far less clear. The currently used cancer risk model by the international radiation protection community states that any increase in radiation exposure proportionately increases the risk of developing cancer. However, this stochastic cancer risk model does not take into account any contribution from non-targeted effects. Nor does it consider the possibility of a bystander mechanism in the induction of genomic instability. This paper reviews the available evidence to date for a possible role for non-targeted effects to contribute to cancer development after exposure to ionising radiation. An evolution in the understanding of the mechanisms driving non-targeted effects after exposure to ionising radiation is critical to determine the true contribution of non-targeted effects on the risk of developing cancer. Such an evolution will likely only be achievable through coordinated multidisciplinary teams combining several fields of study including: genomics, proteomics, cell biology, molecular epidemiology, and traditional epidemiology.
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Carcinogênese , Neoplasias Induzidas por Radiação/etiologia , Efeito Espectador/efeitos da radiação , Instabilidade Genômica/efeitos da radiação , Humanos , Proteção Radiológica , RiscoRESUMO
PURPOSE: Previously published findings have documented increased breast cancer risks associated with the nursing profession. The aim of the present study was to assess whether an increased risk of breast cancer was associated with nursing in a population-based case-control breast cancer study of women in Northeastern Ontario, Canada. METHODS: A total of 1519 women (1380 never-nurses: 716 controls and 664 cases; 139 ever-nurses: 59 controls and 80 cases) were included in the present study. Study participants filled out a detailed questionnaire which included a history of smoking, general health information, breast cancer risk factors, and a detailed occupational history. RESULTS: Ever-nurses were at higher, but nonsignificant risk of breast cancer compared to never-nurses (adjusted OR 1.39, 95 % CI 0.93-2.07). Ever-nurses who worked for longer than 10 years were at a significantly increased risk of breast cancer compared to never-nurses (adjusted OR 1.70, 95 % CI 1.04-2.79). A nonsignificant, but increased risk of breast cancer was observed in ever-nurses who worked full-time compared to never-nurses (OR 1.52, 95 % CI 0.92-2.52), while nurses who worked part-time, or both part-time and full-time were not at increased risk. Ever-nurses who worked in a hospital setting had a significantly increased risk of breast cancer (OR 1.65, 95 % CI 1.04-2.62) compared to never-nurses. CONCLUSIONS: The results indicate that the nurses in the present study population are at increased risk of breast cancer. A prolonged duration of nursing years and prolonged intensity (being a full-time nurse) are factors associated with this increased risk.
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Neoplasias da Mama/epidemiologia , Enfermeiras e Enfermeiros , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Pessoa de Meia-Idade , Ontário , Fatores de Risco , Fumar/efeitos adversos , Inquéritos e QuestionáriosRESUMO
During the first 24 h after removal from incubation, melanoma cells in culture displayed reliable increases in emissions of photons of specific wavelengths during discrete portions of this interval. Applications of specific filters revealed marked and protracted increases in infrared (950 nm) photons about 7 h after removal followed 3 h later by marked and protracted increases in near ultraviolet (370 nm) photon emissions. Specific wavelengths within the visible (400 to 800 nm) peaked 12 to 24 h later. Specific activators or inhibitors for specific wavelengths based upon Cosic's resonant recognition model elicited either enhancement or diminishment of photons at the specific wavelength as predicted. Inhibitors or activators predicted for other wavelengths, even within 10 nm, were less or not effective. There is now evidence for quantitative coupling between the wavelength of photon emissions and intrinsic cellular chemistry. The results are consistent with initial activation of signaling molecules associated with infrared followed about 3 h later by growth and protein-structural factors associated with ultraviolet. The greater-than-expected photon counts compared with raw measures through the various filters, which also function as reflective material to other photons, suggest that photons of different wavelengths might be self-stimulatory and could play a significant role in cell-to-cell communication.
Assuntos
Substâncias Macromoleculares/química , Melanoma Experimental/fisiopatologia , Modelos Teóricos , Fótons , Luz , Melanoma Experimental/química , Células Tumorais Cultivadas , Raios UltravioletaRESUMO
If parameters for lateral diffusion of lipids within membranes are macroscopic metaphors of the angular magnetic moment of the Bohr magneton then the energy emission should be within the visible wavelength for applied ~1 µT magnetic fields. Single or paired digital photomultiplier tubes (PMTs) were placed near dishes of ~1 million B16 mouse melanoma cells that had been removed from incubation. In very dark conditions (10(-11) W/m(2)) different averaged (RMS) intensities between 5 nT and 3.5 µT were applied randomly in 4 min increments. Numbers of photons were recorded directly over or beside the cell dishes by PMTs placed in pairs within various planes. Spectral analyses were completed for photon power density. The peak photon emissions occurred around 1 µT as predicted by the equation. Spectra analyses showed reliable discrete peaks between 0.9 and 1.8 µT but not for lesser or greater intensities; these peak frequencies corresponded to the energy difference of the orbital-spin magnetic moment of the electron within the applied range of magnetic field intensities and the standard solution for Rydberg atoms. Numbers of photons from cooling cells can be modified by applying specific intensities of temporally patterned magnetic fields. There may be a type of "cellular" magnetic moment that, when stimulated by intensity-tuned magnetic fields, results in photon emissions whose peak frequencies reflect predicted energies for fundamental orbital/spin properties of the electron and atomic aggregates with large principal quantum numbers.
Assuntos
Magnetismo , Melanoma Experimental/diagnóstico , Melanoma/diagnóstico , Animais , Lipídeos/química , Campos Magnéticos , Melanoma/patologia , Melanoma Experimental/patologia , Camundongos , Fotoquímica/métodos , Fótons , Processamento de Sinais Assistido por Computador , Software , Espectrofotometria/métodos , Fatores de TempoRESUMO
Exposure to time-varying electromagnetic fields (EMF) has the capacity to influence biological systems. Our results demonstrate that exposure to time-varying EMF modeled after the physiological firing frequency of intercellular calcium waves can inhibit proliferation and induce apoptosis in malignant cells. Single exposure of B16-BL6 cells to a Ca2+ EMF for 40 min reduced the number of viable cells by 50.3%. Cell imaging with acridine orange and ethidium bromide dye revealed substantial cellular apoptosis, preapoptotic cells, nuclear fragmentation, and large spacing between cells in the Ca2+ EMF condition when compared to the control condition. The ability of Ca2+ EMF to influence the proliferation and survival of malignant cells suggests that exposure to specific EMF may function as a potential anticancer therapy.
Assuntos
Sinalização do Cálcio , Melanoma Experimental , Animais , Humanos , Campos Eletromagnéticos , Apoptose , Fatores de TempoRESUMO
Photon emissions were measured at ambient temperature (21°C) in complete darkness once per min from cultures of 10(6) cells during the 12 h following removal from 37°C. The energy of emission was about 10(-20) J/s/cell. Of 8 different cell lines, B16-BL6 (mouse melanoma cells) demonstrated the most conspicuous emission profile. Acridine orange and ethidium bromide indicated the membranes were intact with no indication of (trypan blue) cell necrosis. Treatments with EGF and ionomycin produced rapid early (first 3 h) increases in energy emission while glutamine-free, sodium azide and wortmanin-treated cells showed a general diminishment 3 to 9 h later. The results suggested the most probable origin of the photon emission was the plasma cell membrane. Measures from cells synchronized at the M- and S-phase supported this inference.
Assuntos
Membrana Celular/metabolismo , Animais , Biofísica/métodos , Técnicas de Cultura de Células , Ciclo Celular , Ciclosporina/farmacologia , Fator de Crescimento Epidérmico/metabolismo , Citometria de Fluxo/métodos , Humanos , Indóis/farmacologia , Maleimidas/farmacologia , Melanoma Experimental , Camundongos , Fótons , Temperatura , Fatores de TempoRESUMO
OBJECTIVES: This study examined the activities of tobramycin and bismuth against quorum sensing, virulence factors and biofilms of Pseudomonas aeruginosa by co-encapsulating the agents in liposomes in order to achieve greater delivery of the agents. METHODS: The inhibitory effects of the agents, in either their conventional (free) or vesicle-entrapped (liposomal) formulations, were assessed by measuring the changes in the quorum-sensing signal molecule N-acyl homoserine lactone, pyoverdine, pyocyanin, elastase, protease, chitinase, bacterial attachment and biofilms in vitro. RESULTS: The effectiveness of tobramycin and bismuth was superior when they were co-administered as a liposomal formulation as measured by their ability to attenuate the production of N-acyl homoserine lactone, elastase (P < 0.01), protease (P < 0.05) and chitinase (P < 0.01). In the presence of non-lethal concentrations of free and liposomal tobramycin and bismuth, bacterial attachment was attenuated. Biofilm formation was also attenuated with free tobramycin and bismuth, yet, in the presence of liposomal tobramycin and bismuth, biofilm complexes could form but contained mostly dead bacteria. When established biofilms were treated with higher concentrations, free tobramycin and bismuth killed and detached bacteria, while the liposomal tobramycin and bismuth penetrated and killed bacteria in the cores of the biofilms. CONCLUSIONS: These data suggest that treatment of P. aeruginosa with tobramycin and bismuth, as measured by the changes in quorum sensing, virulence factors and biofilms, is most effective when delivered as a liposomal formulation at a lower concentration compared with the free formulation.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Lipossomos/farmacocinética , Mercaptoetanol/análogos & derivados , Pseudomonas aeruginosa/efeitos dos fármacos , Tobramicina/farmacologia , Fatores de Virulência/antagonistas & inibidores , Proteínas de Bactérias/antagonistas & inibidores , Humanos , Lipossomos/farmacologia , Mercaptoetanol/farmacocinética , Mercaptoetanol/farmacologia , Percepção de Quorum/efeitos dos fármacos , Tobramicina/farmacocinéticaRESUMO
There is a need to synthesize new gene delivery vehicles that can deal with the problems of endosomal escape and nuclear entry. We propose cationic glycopolymer-stabilized gold nanoparticles as an effective gene delivery system. The cationic glyconanoparticles synthesized were revealed to be biocompatible and are resistant to aggregation in physiological conditions. The complexation of DNA to the cationic glyconanoparticles is determined by agarose gel electrophoresis. The localization of the DNA-glyconanoparticles inside the Hela cell line and their mechanism of uptake is studied by confocal microscopy. Finally, the efficacy of the glyconanoparticles as gene delivery vehicles in vitro is studied by their complexation with cyanine fluorescence protein encoded plasmid, and the transfection efficiency is found to be comparable to the commercially available control Lipofectamine 2000.
Assuntos
Carboidratos/química , DNA/administração & dosagem , Nanopartículas/química , Polímeros/química , Transfecção/métodos , Materiais Biocompatíveis , Cátions , Genes Reporter , Células HeLa , Humanos , Farmacocinética , Plasmídeos , Transfecção/normasRESUMO
Flaxseed oil is widely recognized for its exceptional nutritional value, high concentration of fiber-based lignans and large amounts of ω-fatty acids. It is one of a generic group of functional foods that is often taken by cancer patients as a potential treatment. We have examined the anti-cancer effects of flaxseed oil by studying its direct effects on cancer cell growth in vitro. Treatment of a variety of cancer cell lines with flaxseed oil decreased their growth in a dose-dependent manner while non-malignant cell lines showed small increases in cell growth. Cells treated with a mixture of fatty acids, including α-linolenic acid, docosahexaenoic acid, and eicosapentaenoic acid and lignans including enterodiol and enterolactone was also able to decrease the growth of cancer cells. Treatment of B16-BL6 murine melanoma and MCF-7 breast cancer cells with flaxseed oil induced apoptosis as determined by changes in cell morphology, annexin V staining, DNA fragmentation and/or caspase activation. In addition, treatment with flaxseed oil also disrupted mitochondrial function in B16-BL6 and MCF-7 cells. These results indicate that flaxseed oil can specifically inhibit cancer cell growth and induce apoptosis in some cancer cells and suggests it has further potential in anti-cancer therapy.
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BACKGROUND: The frequency of circulating leukocytes has been shown to be a prognostic factor in patients being treated for different types of cancer. In breast cancer, tumor-infiltrating leukocytes may predict patient outcome, but few studies have investigated such associations for circulating leukocytes. PATIENTS AND METHODS: Multiparametric flow cytometry was used to examine the immunophenotypes of circulating peripheral blood mononuclear cells for 88 patients with metastatic breast cancer, which was then correlated to breast cancer-specific survival. Patients had been treated either with high-dose cyclophosphamide-containing regimens (group 1, n = 51 patients) or high-dose paclitaxel-containing regimens (group 2, n = 37 patients). RESULTS: The frequency of peripheral blood CD14+ monocytes indicated prognosis for patients in group 1 (but not group 2), while higher levels of CD11c+ dendritic cells indicated a better prognosis for patients in group 2 (but not group 1). The frequency of a number of different CD4+ or CD8+ T cell subtypes also predicted prognosis for patients in group 2. For example, patients in group 2 with a higher frequency of circulating CD4+ or CD8+ naive T cells (CD45RA+CD95-CD27+CD28+) showed a poorer prognosis. In contrast, T cells were not associated with prognosis for patients in group 1. CONCLUSION: Circulating leukocytes can predict clinical outcome for patients with breast cancer. Prediction of clinical outcome in this cohort of metastatic breast cancer patients was specific to the type of chemotherapy, and this finding is likely to apply to other therapies.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/mortalidade , Células Dendríticas/imunologia , Leucócitos Mononucleares/imunologia , Linfócitos do Interstício Tumoral/imunologia , Monócitos/imunologia , Recidiva Local de Neoplasia/mortalidade , Adulto , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Carboplatina/administração & dosagem , Estudos de Coortes , Ciclofosfamida/administração & dosagem , Células Dendríticas/efeitos dos fármacos , Feminino , Seguimentos , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Linfócitos do Interstício Tumoral/efeitos dos fármacos , Pessoa de Meia-Idade , Mitoxantrona/administração & dosagem , Monócitos/efeitos dos fármacos , Metástase Neoplásica , Recidiva Local de Neoplasia/tratamento farmacológico , Recidiva Local de Neoplasia/imunologia , Recidiva Local de Neoplasia/patologia , Paclitaxel/administração & dosagem , Prognóstico , Taxa de Sobrevida , Tiotepa/administração & dosagem , Vimblastina/administração & dosagemRESUMO
Uncaria tomentosa, commonly known as cat's claw, is a medicinal plant native to Peru, which has been used for decades in the treatment of various inflammatory disorders. Uncaria tomentosa can be used as an antioxidant, has anti-apoptotic properties, and can enhance DNA repair, however it is best know for its anti-inflammatory properties. Treatment with Uncaria tomentosa extracts inhibits the production of the pro-inflammatory cytokine, TNF-alpha, which is a critical mediator of the immune response. In this paper, we showed that treatment of THP-1 monocyte-like cells with Uncaria tomentosa extracts inhibited the MAP kinase signaling pathway and altered cytokine expression. Using ELISA assays, we showed that treatment with Uncaria tomentosa extracts augmented LPS-dependent expression of IL-1beta by 2.4-fold, while inhibiting the LPS-dependent expression of TNF-alpha by 5.5-fold. We also showed that treatment of LPS-stimulated THP-1 cells with Uncaria tomentosa extracts blocked ERK1/2 and MEK1/2 phosphorylation in a dose-dependent manner. These data demonstrate that treatment of THP-1 cells with Uncaria tomentosa extracts has opposite effects on IL-1beta and TNF-alpha secretion, and that these changes may involve effects on the MAP kinase pathway.
Assuntos
Unha-de-Gato/química , Interleucina-1beta/metabolismo , Extratos Vegetais/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Lipopolissacarídeos/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Monócitos/citologia , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Extratos Vegetais/química , Plantas MedicinaisRESUMO
BACKGROUND: Uncaria tomentosa (Willd. ex Schult.) DC (Rubiaceae), known as Cat's Claw or Uña de gato, is a traditionally used medicinal plant native to Peru. Some studies have shown that U. tomentosa can act as an antiapoptotic agent and enhance DNA repair in chemotherapy-treated cells although others have shown that U. tomentosa enhanced apoptosis. OBJECTIVE: To determine if treatment with U. tomentosa can significantly enhance cell death in THP-1 cells exposed to ionizing radiation. MATERIALS AND METHODS: THP-1 monocyte-like cells were treated with ethanolic extracts of U. tomentosa in the presence or absence of bacterial lipopolysaccharide and then exposed to ionizing radiation. Cell proliferation was assessed by MTT and clonogenic assays and the effects on cell cycle measured by flow cytometry and immunoblotting. Changes in cell signaling were determined by immunoblotting and cytokine ELISA and activation of apoptosis measured by caspase activation and DNA fragmentation analysis. RESULTS: Treatment of THP-1 cells with U. tomentosa had a small effect on cell proliferation. However, when the U. tomentosa-pretreated cells were also subjected to 5-9 Gy ionizing radiation, they showed a significant decrease in cell proliferation and increased cellular apoptosis as measured by DNA fragmentation and caspase activation. Treatment with U. tomentosa also decreased the expression of Cyclin E and Cyclin B, key regulators of normal cell cycle progression, and decreased the phosphorylation of various stress-activated, cell survival proteins including p38, ERK, and SAP/JNK kinase. CONCLUSIONS: These results suggest that U. tomentosa could be useful in enhancing cell death following anticancer therapies including ionizing radiation. SUMMARY: Treatment of THP-1 cells with Uncaria tomentosa increases their susceptibility to X-rays. The combination of Uncaria tomentosa and X-ray exposure strongly inhibits cell signaling and promotes apoptosis. Abbreviations Used: LPS: Lipopolysaccharide, TNF: Tumor necrosis factor: IL-1, Interleukin-1: SDS: Sodium dodecylsulphate, TBS: Tris-buffered saline.
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Cellular adhesion molecules are critical components during carcinogenesis and cancer metastasis and contribute to the mechanisms underlying resistance to chemotherapeutic drugs. Since drug resistance is associated with a very poor prognosis for patients with cancer, a better understanding of the role of adhesion molecules could improve patient outcome by identifying novel mechanisms that promote drug resistance. Epigenetic factors, such as cellular adhesion, are shown to promote the resistance of cancers to various chemotherapeutic drugs by altering cellular signalling pathways that activate cellular growth and inhibit apoptosis. In addition, cellular adhesion molecules can provide a means to specifically target more conventional chemotherapy to the unique tumour microenvironment. However, the expression and function of cellular adhesion molecules, and the signals activated by adhesion, are highly interrelated making the development of rational therapies more difficult.
Assuntos
Antineoplásicos/administração & dosagem , Moléculas de Adesão Celular/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Neoplasias/metabolismo , Neoplasias/patologia , Transdução de Sinais/imunologia , Animais , Moléculas de Adesão Celular/fisiologia , Resistencia a Medicamentos Antineoplásicos/imunologia , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Transdução de Sinais/efeitos dos fármacosRESUMO
Tobacco smoking is recognized as a global pandemic resulting in 6 million deaths per year. Despite a variety of anti-smoking products available to aid with tobacco cessation, the majority of people who attempt to quit smoking relapse within 6 months due to the addictive nature of nicotine. An immunotherapy approach could offer a promising treatment option by inducing a potent selective antibody response against nicotine in order to block its distribution to the brain and its addictive effects in the central nervous system. Our nicotine vaccine candidate was administered intranasally using the Neisseria meningitidis serogroup B Adjuvant Finlay Proteoliposome 1 (AFPL1) as a part of the delivery system. This system was designed to generate a robust immune response by stimulating IL-1ß production through Toll-like receptor 4 (TLR4), a potent mechanism for mucosal immunity. The vaccine induced high antibody titers in mice sera in addition to inducing mucosal antibodies. The efficacy of our vaccine was demonstrated using in vivo challenge experiments with radioactive [(3)H]-nicotine, followed by an analysis of nicotine distribution in the lung, liver, blood and brain. Our results were encouraging as the nicotine concentration in the brain tissue of mice vaccinated with our candidate vaccine was four times lower than in non-vaccinated controls; suggesting that the anti-nicotine antibodies were able to block nicotine from crossing the blood brain barrier. In summary, we have developed a novel nicotine vaccine for the treatment of tobacco addiction by intranasal administration and also demonstrated that the AFPL1 can be used as a potential adjuvant for this vaccine design.
RESUMO
The anti-malarial drug chloroquine (CQ) is also thought to be a potential radiation sensitizer. To gain a better understanding of how the lysomotropic CQ can potentiate the effects of ionizing radiation, we investigated the effects of CQ on lysosomal and mitochondrial membrane stability, the subcellular localization of ceramide, plasma membrane permeability, and the mode of cell death in response to irradiation. We found that CQ accumulated in the lysosomes and thus lysosomal volumes increased. As a result, both the lysosomal and plasma membranes were destabilized. After 7 Gy irradiation, most ceramide was associated with the lysosomes in the cells treated with CQ but not in the CQ-untreated control. The elevated levels of ceramide in the lysosomes of the CQ-treated cells appeared to further destabilize the lysosomal and plasma membranes of the cell. Both CQ-treated and -untreated cells had approximately the same rate of cell death by apoptosis after 7 Gy irradiation (P > 0.05, ns). However, in contrast to the CQ-untreated control, the CQ-treated cells underwent massive cell death by necrosis at 24-48 h after irradiation (P < 0.05). Taken together, our data support the idea that the increase in cytotoxic effects by the combination of CQ and radiation is due to radiation-mediated apoptosis and CQ-mediated necrosis.
Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/patologia , Cloroquina/administração & dosagem , Membranas Intracelulares/efeitos dos fármacos , Membranas Intracelulares/efeitos da radiação , Lisossomos/efeitos dos fármacos , Lisossomos/efeitos da radiação , Tolerância a Radiação/efeitos dos fármacos , Apoptose/efeitos da radiação , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Relação Dose-Resposta à Radiação , Humanos , Membranas Intracelulares/patologia , Lisossomos/patologia , Fluidez de Membrana/efeitos dos fármacos , Fluidez de Membrana/efeitos da radiação , Necrose/patologia , Doses de Radiação , Radiossensibilizantes/administração & dosagemRESUMO
Electromagnetic field (EMF) exposures affect many biological systems. The reproducibility of these effects is related to the intensity, duration, frequency, and pattern of the EMF. We have shown that exposure to a specific time-varying EMF can inhibit the growth of malignant cells. Thomas-EMF is a low-intensity, frequency-modulated (25-6 Hz) EMF pattern. Daily, 1 h, exposures to Thomas-EMF inhibited the growth of malignant cell lines including B16-BL6, MDA-MB-231, MCF-7, and HeLa cells but did not affect the growth of non-malignant cells. Thomas-EMF also inhibited B16-BL6 cell proliferation in vivo. B16-BL6 cells implanted in syngeneic C57b mice and exposed daily to Thomas-EMF produced smaller tumours than in sham-treated controls. In vitro studies showed that exposure of malignant cells to Thomas-EMF for > 15 min promoted Ca(2+) influx which could be blocked by inhibitors of voltage-gated T-type Ca(2+) channels. Blocking Ca(2+) uptake also blocked Thomas-EMF-dependent inhibition of cell proliferation. Exposure to Thomas-EMF delayed cell cycle progression and altered cyclin expression consistent with the decrease in cell proliferation. Non-malignant cells did not show any EMF-dependent changes in Ca(2+) influx or cell growth. These data confirm that exposure to a specific EMF pattern can affect cellular processes and that exposure to Thomas-EMF may provide a potential anti-cancer therapy.