RESUMEN
Melanoma is the most lethal form of skin cancer, with a worldwide increase in incidence. Despite the increased overall survival of metastatic melanoma patients given recent advances in targeted and immunotherapy, it still has a poor prognosis and available treatment options carry diverse severe side effects. Polysaccharides from seaweed have been shown to exert antitumor activities. Here we show in vitro and in vivo antitumor activities of a sulfated homogalactan (named 3G4S) from Codium isthmocladum seaweed in the B16-F10 murine melanoma cell line. 3G4S did not induce cytotoxicity or proliferation changes; however, it was able to reduce solid tumor growth and metastasis, while not inducing side effects in mice. B16-F10 cells traits related to the metastatic cascade were also impaired by 3G4S, reducing cell invasion, colony-forming capacity and membrane glycoconjugates. Therefore, 3G4S shows promising antitumor activities without the commonly associated drawbacks of cancer treatments and can be further explored.
Asunto(s)
Galactanos/farmacología , Tecnología Química Verde , Melanoma Experimental/prevención & control , Algas Marinas/química , Sulfatos/química , Animales , Apoptosis , Proliferación Celular , Femenino , Humanos , Masculino , Melanoma Experimental/secundario , Ratones , Ratones Endogámicos C57BL , Células Tumorales CultivadasRESUMEN
Melanoma is a form of skin cancer with high mortality owing to its fast progression and metastatic capacity. The treatments available nowadays are only palliative in advanced stages of the disease. Thus, alternative therapies for cancer treatment are in demand, and molecules from natural sources, such as polysaccharides, could represent new possible therapeutic approaches. Polysaccharides of freshwater and marine algae with biological activities, such as antitumor properties, are greatly reported in the scientific literature. In the present study, a sulfated heterorhamnan obtained from the green seaweed Gayralia brasiliensis (Gb1 fraction) was chemically characterized and its biological activities in the B16-F10 murine melanoma cell line were evaluated. The Gb1 polysaccharidic fraction tested concentrations presented low or absence of cytotoxicity to B16-F10 cells and neither cell proliferation nor cell cycle were altered. Interestingly, Gb1 treatment decreased B16-F10 cells migration and invasion capabilities and CD44 labeling, showing to be a promising compound for further in vitro and in vivo antitumor studies.
Asunto(s)
Chlorophyta/química , Desoxiazúcares/farmacología , Mananos/farmacología , Melanoma/tratamiento farmacológico , Animales , Línea Celular Tumoral , Movimiento Celular , Desoxiazúcares/toxicidad , Mananos/toxicidad , Ratones , Invasividad Neoplásica , SulfatosAsunto(s)
Hematología/educación , Investigación Biomédica , Brasil , Creación de Capacidad , Humanos , Internado y Residencia , PobrezaRESUMEN
A heteropolysaccharide was isolated by cold aqueous extraction from edible mushroom Pleurotus eryngii ("King Oyster") basidiocarps and its biological properties were evaluated. Structural assignments were carried out using mono- and bidimensional NMR spectroscopy, monosaccharide composition, and methylation analyses. A mannogalactan having a main chain of (1â6)-linked α-d-galactopyranosyl and 3-O-methyl-α-d-galactopyranosyl residues, both partially substituted at OH-2 by ß-d-Manp (MG-Pe) single-unit was found. Biological effects of mannogalactan from P. eryngii (MG-Pe) were tested against murine melanoma cells. MG-Pe was non-cytotoxic, but reduced in vitro melanoma cells invasion. Also, 50mg/kg MG-Pe administration to melanoma-bearing C57BL/6 mice up to 10days decreased in 60% the tumor volume compared to control. Additionally, no changes were observed when biochemical profile, complete blood cells count (CBC), organs, and body weight were analyzed. Mg-Pe was shown to be a promising anti-melanoma molecule capable of switching melanoma cells to a non-invasive phenotype with no toxicity to melanoma-bearing mice.
Asunto(s)
Polisacáridos Fúngicos/farmacología , Galactanos/farmacología , Melanoma/tratamiento farmacológico , Pleurotus/química , Animales , Cuerpos Fructíferos de los Hongos/química , Espectroscopía de Resonancia Magnética , Ratones , Ratones Endogámicos C57BLRESUMEN
Macrophages are cells of high plasticity and can act in different ways to ensure that the appropriate immune response remains controlled. This study shows the effects of the C-type Bothrops jararacussu venom lectin (BJcuL) on the activation of human macrophages derived from the U937 cell line. BJcuL binds on the cell surface, and this event is inhibited by its specific carbohydrate. It induced phagocytosis and production of H2O2, and expression of antigen presentation molecules. It also enhanced the production of TNF-α, GM-CSF and IL-6 by macrophages and indirectly induced T cells to an increased production of TNF-α, IFN-γ and IL-6 in the presence of LPS. Our results suggest that BJcuL can modulate macrophage functional activation towards an M1 state.
Asunto(s)
Venenos de Crotálidos/toxicidad , Macrófagos/efectos de los fármacos , Técnicas de Cocultivo , Citocinas/metabolismo , Humanos , Peróxido de Hidrógeno/metabolismo , Lectinas Tipo C , Lipopolisacáridos , Macrófagos/metabolismo , Macrófagos/fisiología , Monocitos/citología , Fagocitosis/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismo , Células U937RESUMEN
It has been hypothesized that oils containing high levels of omega-3 polyunsaturated fatty acids, such as canola and fish oil, could counteract some of the adverse effects induced by phthalates. In the present study, the influence of different oily vehicles on di-butyl phthalate (DBP)-induced testicular toxicity and lipid profile was investigated. Pregnant Wistar rats were treated by oral gavage from gestation days 13 to 20 with DBP (500 mg/kg/day) diluted in three different vehicles: corn, canola or fish oil. Male fetuses were analyzed on gestation day 20. DBP exposure lowered intratesticular testosterone levels and anogenital distance, regardless of the vehicle used. The percentage of seminiferous cords containing multinucleated gonocytes and cord diameter was increased in DBP-exposed groups, compared with vehicle controls, with no difference between the three DBP-exposed groups. Clustering of Leydig cells was seen in all DBP groups. Lipid profile indicated that administration of canola and fish oil can increase the content of omega-3 fatty acids in rat testis. However, content of omega-3 was diminished in DBP-treated groups. Overall, our results indicate that different oily vehicles did not alter fetal rat testicular toxicity induced by a high DBP dose.
Asunto(s)
Dibutil Ftalato/toxicidad , Disruptores Endocrinos/toxicidad , Ácidos Grasos Omega-3/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Exposición Materna/efectos adversos , Vehículos Farmacéuticos/metabolismo , Testículo/efectos de los fármacos , Animales , Aceite de Maíz/química , Aceite de Maíz/metabolismo , Dibutil Ftalato/administración & dosificación , Disruptores Endocrinos/administración & dosificación , Contaminantes Ambientales/administración & dosificación , Contaminantes Ambientales/toxicidad , Ácidos Grasos Monoinsaturados/química , Ácidos Grasos Monoinsaturados/metabolismo , Ácidos Grasos Omega-3/química , Femenino , Desarrollo Fetal/efectos de los fármacos , Aceites de Pescado/química , Aceites de Pescado/metabolismo , Masculino , Vehículos Farmacéuticos/química , Plastificantes/administración & dosificación , Plastificantes/toxicidad , Embarazo , Aceite de Brassica napus , Ratas , Procesos de Determinación del Sexo/efectos de los fármacos , Testículo/embriología , Testículo/metabolismo , Testosterona/metabolismoRESUMEN
We have examined the role of cell surface glycosaminoglycans in cell division: adhesion and proliferation of Chinese hamster ovary (CHO) cells. We used both wild-type (CHO-K1) cells and a mutant (CHO-745) which is deficient in the synthesis of proteoglycans due to lack of activity of xylosyl transferase. Using different amounts of wild-type and mutant cells, little adhesion was observed in the presence of laminin and type I collagen. However, when fibronectin or vitronectin was used as substrate, there was an enhancement in the adhesion of wild-type and mutant cells. Only CHO-K1 cells showed a time-dependent adhesion on type IV collagen. These results suggest that the two cell lines present different adhesive profiles. Several lines of experimental evidence suggest that heparan sulfate proteoglycans play a role in cell adhesion as positive modulators of cell proliferation and as key participants in the process of cell division. Proliferation and cell cycle assays clearly demonstrate that a decrease in the amount of glycosaminoglycans does not inhibit the proliferation of mutant CHO-745 cells when compared to the wild type CHO-K1, in agreement with the findings that both CHO-K1 and CHO-745 cells take 8 h to enter the S phase
Asunto(s)
Animales , Cricetinae , Células CHO/citología , Matriz Extracelular/fisiología , Proteoglicanos de Heparán Sulfato/fisiología , Adhesión Celular/fisiología , División Celular , Colágeno/fisiología , Fibronectinas/fisiología , Laminina/fisiología , Vitronectina/fisiologíaRESUMEN
We have examined the role of cell surface glycosaminoglycans in cell division: adhesion and proliferation of Chinese hamster ovary (CHO) cells. We used both wild-type (CHO-K1) cells and a mutant (CHO-745) which is deficient in the synthesis of proteoglycans due to lack of activity of xylosyl transferase. Using different amounts of wild-type and mutant cells, little adhesion was observed in the presence of laminin and type I collagen. However, when fibronectin or vitronectin was used as substrate, there was an enhancement in the adhesion of wild-type and mutant cells. Only CHO-K1 cells showed a time-dependent adhesion on type IV collagen. These results suggest that the two cell lines present different adhesive profiles. Several lines of experimental evidence suggest that heparan sulfate proteoglycans play a role in cell adhesion as positive modulators of cell proliferation and as key participants in the process of cell division. Proliferation and cell cycle assays clearly demonstrate that a decrease in the amount of glycosaminoglycans does not inhibit the proliferation of mutant CHO-745 cells when compared to the wild type CHO-K1, in agreement with the findings that both CHO-K1 and CHO-745 cells take 8 h to enter the S phase.
Asunto(s)
Células CHO/citología , Matriz Extracelular/fisiología , Proteoglicanos de Heparán Sulfato/fisiología , Animales , Células CHO/fisiología , Adhesión Celular/fisiología , División Celular/fisiología , Colágeno/fisiología , Cricetinae , Fibronectinas/fisiología , Laminina/fisiología , Vitronectina/fisiologíaRESUMEN
The effect of a cyclic octaphenol-octasulfonic acid (GL-522-Y-1), upon the synthesis of a heparan sulfate proteoglycan synthesized by endothelial cells (rabbit aorta and human umbilical vein) were studied. The cells were exposed to the compounds at various concentrations for different periods of time and the synthesized heparan sulfates analyzed by a combination of agarose gel electrophoresis and enzymatic degradation. The GL-522-Y-1, like heparin, change the sulfation pattern and stimulate two- to three-fold the synthesis of heparan sulfate proteoglycan secreted by rabbit and human endothelial cells in culture. GL-522-Y-1, besides being 100 times more active than heparin, also produces a significant enhancement of cell surface heparan sulfate in human vein endothelial cells. The effect of GL-522-Y-1 is completely abolished by methylation or acetylation of its free hydroxyl groups. Both heparin and GL-522-Y-1 have high affinity for a 47-kDa protein present at the surface of endothelial cells. These and other results lead us to speculate that the antithrombotic activity of heparin and GL522 "in vivo" could be related, at least in part, to the increased production of the heparan sulfate proteoglycan by endothelial cells.
Asunto(s)
Anticoagulantes/farmacología , Endotelio Vascular/efectos de los fármacos , Proteoglicanos de Heparán Sulfato/biosíntesis , Heparina/farmacología , Proteínas de la Membrana/metabolismo , Inhibidores de Agregación Plaquetaria/farmacología , Polímeros/farmacología , Ácidos Sulfónicos/farmacología , Animales , Anticoagulantes/metabolismo , Aorta , Relación Dosis-Respuesta a Droga , Endotelio Vascular/química , Endotelio Vascular/citología , Proteoglicanos de Heparán Sulfato/química , Heparina/metabolismo , Humanos , Proteínas de la Membrana/química , Estructura Molecular , Peso Molecular , Inhibidores de Agregación Plaquetaria/metabolismo , Polímeros/metabolismo , Unión Proteica , Conejos , Ácidos Sulfónicos/metabolismo , Venas UmbilicalesRESUMEN
The anticlotting and antithrombotic activities of heparin, heparan sulfate, low molecular weight heparins, heparin and heparin-like compounds from various sources used in clinical practice or under development are briefly reviewed. Heparin isolated from shrimp mimics the pharmacological activities of low molecular weight heparins. A heparan sulfate from Artemia franciscana and a dermatan sulfate from tuna fish show a potent heparin cofactor II activity. A heparan sulfate derived from bovine pancreas has a potent antithrombotic activity in an arterial and venous thrombosis model with a negligible activity upon the serine proteases of the coagulation cascade. It is suggested that the antithrombotic activity of heparin and other antithrombotic agents is due at least in part to their action on endothelial cells stimulating the synthesis of an antithrombotic heparan sulfate.
Asunto(s)
Humanos , Animales , Bovinos , Anticoagulantes/farmacología , Endotelio Vascular/citología , Fibrinolíticos/farmacología , Heparina/farmacología , Heparitina Sulfato/farmacología , Anticoagulantes/química , Anticoagulantes/metabolismo , Crustáceos , Fibrinolíticos/química , Fibrinolíticos/metabolismo , Glicosaminoglicanos/metabolismo , Glicosaminoglicanos/farmacología , Heparina de Bajo-Peso-Molecular/química , Heparina de Bajo-Peso-Molecular/metabolismo , Heparina de Bajo-Peso-Molecular/farmacología , Heparina/metabolismo , Heparitina Sulfato/biosíntesis , AtúnRESUMEN
The effect of brown spider (Loxosceles intermedia) venom on endothelial cells was investigated in vivo and in vitro. Morphological and ultrastructural observations by light microscopy and transmission electron microscopy showed that the venom acts in vivo upon vessel endothelial cells of rabbits that were intradermally injected, evoking vessel instability, cytoplasmic endothelial cell vacuolization, and blebs. Likewise, treatment of rabbit endothelial cells in culture with the venom led to loss of adhesion of the cells to the substrate. Endothelial cells in culture were metabolically radiolabeled with sodium [35S]-sulfate and the sulfated compounds (proteoglycans and sulfated proteins) from medium, cell surface, and extracellular matrix (ECM) were analyzed. Agarose gel electrophoresis and SDS-PAGE showed that the venom is active on the ECM and on cell surface proteoglycans, shedding these molecules into the culture medium. In addition, when purified heparan sulfate proteoglycan (HSPG) and purified laminin-entactin (LN/ET) complex were incubated with the venom we observed a partial degradation of the protein core of HSPG as well as the hydrolysis of entactin. The above results suggest that the L. intermedia venom has a deleterious effect on the endothelium of vessels both in vivo and in culture, removing important constituents such as HSPG and entactin that are involved in the adhesion of endothelial cells and of subendothelial ECM organization.
Asunto(s)
Citotoxinas/farmacología , Endotelio Vascular/citología , Hidrolasas Diéster Fosfóricas/farmacología , Venenos de Araña/farmacología , Animales , Membrana Basal/química , Línea Celular , Células Cultivadas , Endotelio Vascular/efectos de los fármacos , Proteoglicanos de Heparán Sulfato/metabolismo , Glicoproteínas de Membrana/efectos de los fármacos , Glicoproteínas de Membrana/metabolismo , Microscopía Electrónica , ConejosRESUMEN
The anticlotting and antithrombotic activities of heparin, heparan sulfate, low molecular weight heparins, heparin and heparin-like compounds from various sources used in clinical practice or under development are briefly reviewed. Heparin isolated from shrimp mimics the pharmacological activities of low molecular weight heparins. A heparan sulfate from Artemia franciscana and a dermatan sulfate from tuna fish show a potent heparin cofactor II activity. A heparan sulfate derived from bovine pancreas has a potent antithrombotic activity in an arterial and venous thrombosis model with a negligible activity upon the serine proteases of the coagulation cascade. It is suggested that the antithrombotic activity of heparin and other antithrombotic agents is due at least in part to their action on endothelial cells stimulating the synthesis of an antithrombotic heparan sulfate.
Asunto(s)
Anticoagulantes/farmacología , Endotelio Vascular/efectos de los fármacos , Fibrinolíticos/farmacología , Heparina/farmacología , Heparitina Sulfato/farmacología , Animales , Anticoagulantes/química , Anticoagulantes/metabolismo , Bovinos , Crustáceos , Endotelio Vascular/citología , Fibrinolíticos/química , Fibrinolíticos/metabolismo , Glicosaminoglicanos/metabolismo , Glicosaminoglicanos/farmacología , Heparina/química , Heparina/metabolismo , Heparina de Bajo-Peso-Molecular/química , Heparina de Bajo-Peso-Molecular/metabolismo , Heparina de Bajo-Peso-Molecular/farmacología , Heparitina Sulfato/biosíntesis , Humanos , Relación Estructura-Actividad , AtúnRESUMEN
Fucans, a family of sulfated polysaccharides present in brown seaweed, have several biological activities. Their use as drugs would offer the advantage of no potential risk of contamination with viruses or particles such as prions. A fucan prepared from Spatoglossum schröederi was tested as a possible inhibitor of cell-matrix interactions using wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). The effect of this polymer on adhesion properties with specific extracellular matrix components was studied using several matrix proteins as substrates for cell attachment. Treatment with the polymer inhibited the adhesion of fibronectin to both CHO-K1 (2 x 10(5))()and CHO-745 (2 x 10(5) and 5 x 10(5)) cells. No effect was detected with laminin, using the two cell types. On the other hand, adhesion to vitronectin was inhibited in CHO-K1 cells and adhesion to type I collagen was inhibited in CHO-745 cells. In spite of this inhibition, the fucan did not affect either cell proliferation or cell cycle. These results demonstrate that this polymer is a new anti-adhesive compound with potential pharmacological applications
Asunto(s)
Animales , Cricetinae , Anticoagulantes/química , Adhesión Celular/efectos de los fármacos , Proteínas de la Matriz Extracelular/química , Polisacáridos/química , Algas Marinas/química , Anticoagulantes/farmacología , Ciclo Celular , División Celular/efectos de los fármacos , Células CHO , Proteínas de la Matriz Extracelular/antagonistas & inhibidores , Polisacáridos/farmacología , Timidina/metabolismoRESUMEN
Fucans, a family of sulfated polysaccharides present in brown seaweed, have several biological activities. Their use as drugs would offer the advantage of no potential risk of contamination with viruses or particles such as prions. A fucan prepared from Spatoglossum schröederi was tested as a possible inhibitor of cell-matrix interactions using wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). The effect of this polymer on adhesion properties with specific extracellular matrix components was studied using several matrix proteins as substrates for cell attachment. Treatment with the polymer inhibited the adhesion of fibronectin to both CHO-K1 (2 x 10(5)) and CHO-745 (2 x 10(5) and 5 x 10(5)) cells. No effect was detected with laminin, using the two cell types. On the other hand, adhesion to vitronectin was inhibited in CHO-K1 cells and adhesion to type I collagen was inhibited in CHO-745 cells. In spite of this inhibition, the fucan did not affect either cell proliferation or cell cycle. These results demonstrate that this polymer is a new anti-adhesive compound with potential pharmacological applications.