RESUMEN
Natural substances are strategic candidates for drug development in cancer research. Marine-derived molecules are of special interest due to their wide range of biological activities and sustainable large-scale production. Melanoma is a type of skin cancer that originates from genetic mutations in melanocytes. BRAF, RAS, and NF1 mutations are described as the major melanoma drivers, but approximately 20% of patients lack these mutations and are included in the triple wild-type (tripleWT) classification. Recent advances in targeted therapy directed at driver mutations along with immunotherapy have only partially improved patients' overall survival, and consequently, melanoma remains deadly when in advanced stages. Fucose-containing sulfated polysaccharides (FCSP) are potential candidates to treat melanoma; therefore, we investigated Fucan A, a FCSP from Spatoglossum schröederi brown seaweed, in vitro in human melanoma cell lines presenting different mutations. Up to 72 h Fucan A treatment was not cytotoxic either to normal melanocytes or melanoma cell lines. Interestingly, it was able to impair the tripleWT CHL-1 cell proliferation (57%), comparable to the chemotherapeutic cytotoxic drug cisplatin results, with the advantage of not causing cytotoxicity. Fucan A increased CHL-1 doubling time, an effect attributed to cell cycle arrest. Vascular mimicry, a close related angiogenesis process, was also impaired (73%). Fucan A mode of action could be related to gene expression modulation, in special ß-catenin downregulation, a molecule with protagonist roles in important signaling pathways. Taken together, results indicate that Fucan A is a potential anticancer molecule and, therefore, deserves further investigation.
Asunto(s)
Antineoplásicos , Melanoma , Phaeophyceae , Humanos , Fucosa , Sulfatos/farmacología , Melanoma/tratamiento farmacológico , Línea Celular , Polisacáridos/farmacología , Polisacáridos/metabolismo , Antineoplásicos/farmacologíaRESUMEN
BACKGROUND: Silver nanoparticles (AgNP) are largely used in nanotechnological products, but the real risks for human and environment are still poorly understood if we consider the effects of mixtures of AgNP and environmental contaminants, such as non-essential metals. METHODS: The aim of the present study was to investigate the cytotoxicity and toxicological interaction of AgNP (1-4 nm, 0.36 and 3.6 µg mL-1) and cadmium (Cd, 1 and 10 µM) mixtures. The murine macrophage cell line RAW 264.7 was used as a model. RESULTS: Effects were observed after a few hours (4 h) on reactive oxygen species (ROS) and became more pronounced after 24 h-exposure. Cell death occurred by apoptosis, and loss of cell viability (24 h-exposure) was preceded by increases of ROS levels and DNA repair foci, but not of NO levels. Co-exposure potentiated some effects (decrease of cell viability and increase of ROS and NO levels), indicating toxicological interaction. CONCLUSION: These effects are important findings that must be better investigated, since the interaction of Cd with AgNP from nanoproducts may impair the function of macrophages and represent a health risk for humans.
Asunto(s)
Nanopartículas del Metal , Plata , Animales , Cadmio/toxicidad , Cloruro de Cadmio , Línea Celular , Supervivencia Celular , Humanos , Macrófagos , Nanopartículas del Metal/toxicidad , Ratones , Especies Reactivas de Oxígeno , Plata/toxicidadRESUMEN
Gold nanoparticle (AuNP)-based systems have been extensively investigated as diagnostic and therapeutic agents due to their tunable properties and easy surface functionalization. Upon cell uptake, AuNPs present an inherent cell impairment potential based on organelle and macromolecules damage, leading to cell death. Such cytotoxicity is concentration-dependent and completely undesirable, especially if unspecific. However, under non-cytotoxic concentrations, internalized AuNPs could potentially weaken cells and act as antitumor agents. Therefore, this study aimed to investigate the antitumor effect of ultrasmall AuNPs (~3 nm) stabilized by the anionic polysaccharide gum arabic (GA-AuNPs). Other than intrinsic cytotoxicity, the focus was downregulation of cancer hallmarks of aggressive tumors, using a highly metastatic model of melanoma. We first demonstrated that GA-AuNPs showed excellent stability under biological environment. Non-cytotoxic concentrations to seven different cell lines, including tumorigenic and non-tumorigenic cells, were determined by standard 2D in vitro assays. Gold concentrations ≤ 2.4 mg L-1 (16.5 nM AuNPs) were non-cytotoxic and therefore chosen for further analyses. Cells exposed to GA-AuNPs were uptaken by melanoma cells through endocytic processes. Next we described remarkable biological properties using non-cytotoxic concentrations of this nanomaterial. Invasion through an extracellular matrix barrier as well as 3D growth capacity (anchorage-independent colony formation and spheroids growth) were negatively affected by 2.4 mg L-1 GA-AuNPs. Additionally, exposed spheroids showed morphological changes, suggesting that GA-AuNPs could penetrate into the preformed tumor and affect its integrity. All together these results demonstrate that side effects, such as cytotoxicity, can be avoided by choosing the right concentration, nevertheless, preserving desirable effects such as modulation of key tumor cell malignancy features.
Asunto(s)
Antineoplásicos/farmacología , Movimiento Celular/efectos de los fármacos , Compuestos de Oro/farmacología , Melanoma Experimental/tratamiento farmacológico , Nanopartículas del Metal , Neoplasias Cutáneas/tratamiento farmacológico , Animales , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/toxicidad , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Estabilidad de Medicamentos , Endocitosis , Compuestos de Oro/química , Compuestos de Oro/metabolismo , Compuestos de Oro/toxicidad , Goma Arábiga/química , Humanos , Melanoma Experimental/metabolismo , Melanoma Experimental/patología , Nanopartículas del Metal/química , Nanopartículas del Metal/toxicidad , Ratones , Nanomedicina , Invasividad Neoplásica , Metástasis de la Neoplasia , Tamaño de la Partícula , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patologíaRESUMEN
Metastasis is responsible for the majority of deaths among patients with malignant melanoma. Despite recent advances, the majority of current and modern therapies are ineffective and/or financially unfeasible. Thus, in this study, we investigated two lowcost highlydiluted natural complexes (HDNCs) that have been shown to be effective against malignant melanoma in a murine model in vivo. The aim of this study was to determine the mechanisms through which these HDNCs directly affect melanoma cells, either alone or in an artificial tumor microenvironment, suppressing the metastatic phenotype, thus explaining previous in vivo effects. For this purpose, HDNC in vitro treatments of B16F10 melanoma cells, alone or in coculture with Balb/3T3 fibroblasts, were carried out. Molecular biology techniques and standard functional assays were used to assess the changes in molecule expression and in cell behaviors related to the metastatic phenotype. Melanoma progression features were found to be regulated by HDNCs. Molecules related to cell adhesion (Ncadherin, ß1integrin and CD44), and migration, extracellular matrix remodeling and angiogenesis were modulated. The cell migratory, invasive and clonogenic capacities were reduced by the HDNCs. No loss of cell proliferation or viability were observed. On the whole, the findings of this study indicate that HDNCs directly reprogram, molecularly and functionally, melanoma cells in vitro, modulating their metastatic phenotype. Such findings are likely to be responsible for the attenuation of tumor growth and lung colonization previously observed in vivo.
Asunto(s)
Antígenos CD/metabolismo , Cadherinas/metabolismo , Receptores de Hialuranos/metabolismo , Integrina beta1/metabolismo , Melanoma/metabolismo , Extractos Vegetales/farmacología , Neoplasias Cutáneas/metabolismo , Animales , Células 3T3 BALB , Adhesión Celular , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Ensayos de Selección de Medicamentos Antitumorales , Regulación Neoplásica de la Expresión Génica , Humanos , Melanoma/tratamiento farmacológico , Ratones , Metástasis de la Neoplasia , Extractos Vegetales/química , Plantas/química , Neoplasias Cutáneas/tratamiento farmacológico , Melanoma Cutáneo MalignoRESUMEN
[This corrects the article DOI: 10.1039/C5RA24858B.].
RESUMEN
BACKGROUND: M1 is a homeopathic medicine with immunostimulatory properties used mainly by cancer patients to complement current therapies. Metastatic melanoma is a skin-originated form of cancer without a single therapy able to produce high rate and sustained responses, which attracts the use of complementary therapies such as M1. However, M1's anti-melanoma effects remain to be pre-clinically demonstrated. Therefore in the present work, we utilized a pulmonary metastatic melanoma model and a subcutaneous melanoma growth model to investigate the potential benefits of treatment with M1. METHODS: C57BL/6 mice were injected intravenously or subcutaneously with B16F10 mouse melanoma cells. After 24 h, mice were treated with either M1 or vehicle (water) for 14 days, euthanized and harvested for multi-parameter pulmonary and tumor analyses. RESULTS: Mice treated with M1 had significantly lower tumor burden in the lungs and subcutaneous tissue than control mice. Furthermore, tumors were impaired in proliferation and tumor related angiogenesis by the inhibition of myeloid derived suppressor cells (MDSC) positive for angiotensin II type 1 receptor (AT1R). CONCLUSION: Altogether these data suggest M1 is an efficient candidate for melanoma therapy to be considered for future clinic studies as this study is the first supporting the idea that melanoma patients may benefit with the treatment. The treatment with M1 provides advantages considering the highly-diluted properties and a cost effective alternative to costly chemotherapeutic approaches with, if any, lower toxicity.
Asunto(s)
Homeopatía/métodos , Materia Medica/uso terapéutico , Melanoma/tratamiento farmacológico , Melanoma/prevención & control , Terapia Respiratoria/métodos , Animales , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BLRESUMEN
Leishmania disease expression has been linked to IL-10. In this study, we investigated the regulation of IL-10 production by macrophages infected with Leishmania donovani. Infection of either murine or human macrophages brought about selective phosphorylation of Akt-2 in a PI3K-dependent manner. These events were linked to phosphorylation and inactivation of glycogen synthase kinase-3ß (GSK-3ß) at serine 9, as the latter was abrogated by inhibition of either PI3K or Akt. One of the transcription factors that is negatively regulated by GSK-3ß is CREB, which itself positively regulates IL-10 expression. Infection of macrophages with leishmania induced phosphorylation of CREB at serine 133, and this was associated with enhanced CREB DNA binding activity and induction of IL-10. Similar to phosphorylation of GSK-3ß, both phosphorylation of CREB at serine 133 and CREB DNA binding activity were abrogated in cells treated with inhibitors of either PI3K or Akt prior to infection. Furthermore, disruption of this pathway either by inhibition of Akt or by overexpression of GSK-3ß markedly attenuated IL-10 production in response to leishmania. Thus, GSK-3ß negatively regulates myeloid cell IL-10 production in response to leishmania. Switching off GSK-3ß promotes disease pathogenesis.