RESUMEN
PURPOSE: Cancer cachexia contributes significantly to morbidity and mortality in individuals with cancer. Currently, the mechanisms contributing to the development of cachexia are largely unknown, leading to a paucity of treatment and prevention options. Animal models are necessary in determining causal mechanisms and in testing potential treatments. While the Yoshida sarcoma has been utilized for more than 50 years, the cachexia syndrome produced by this model has not been well characterized in the literature. METHODS: Tumor allografts were subcutaneously implanted in male Sprague Dawley rats (n = 16) and allowed to grow for 23 days. Control animals (n = 16) received a sham surgery. All rats were monitored daily for the presence of hallmark cachexia symptoms. RESULTS: The results demonstrate the presence of decreased body weight gain, as well as lower levels of body adiposity and skeletal muscle mass, in tumor-bearing animals, as compared to controls. CONCLUSIONS: While a large tumor burden was reached, the extent of cachexia was similar to that which is observed in many individuals with cancer cachexia. Future experiments utilizing this model are encouraged to identify mechanisms and effective treatment and prevention strategies.
Asunto(s)
Caquexia/metabolismo , Caquexia/patología , Modelos Animales de Enfermedad , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/patología , Animales , Glucemia/metabolismo , Caquexia/sangre , Caquexia/etiología , Ingestión de Alimentos , Xenoinjertos , Insulina/sangre , Masculino , Ratas , Ratas Sprague-Dawley , Sarcoma de Yoshida/sangre , Pérdida de PesoRESUMEN
BACKGROUND: Cachexia is a wasting condition that manifests in several types of cancer, and the main characteristic is the profound loss of muscle mass. METHODS: The Yoshida AH-130 tumor model has been used and the samples have been analyzed using transmission electronic microscopy, real-time PCR and Western blot techniques. RESULTS: Using in vivo cancer cachectic model in rats, here we show that skeletal muscle loss is accompanied by fiber morphologic alterations such as mitochondrial disruption, dilatation of sarcoplasmic reticulum and apoptotic nuclei. Analyzing the expression of some factors related to proteolytic and thermogenic processes, we observed in tumor-bearing animals an increased expression of genes involved in proteolysis such as ubiquitin ligases Muscle Ring Finger 1 (MuRF-1) and Muscle Atrophy F-box protein (MAFBx). Moreover, an overexpression of both sarco/endoplasmic Ca(2+)-ATPase (SERCA1) and adenine nucleotide translocator (ANT1), both factors related to cellular energetic efficiency, was observed. Tumor burden also leads to a marked decreased in muscle ATP content. CONCLUSIONS: In addition to muscle proteolysis, other ATP-related pathways may have a key role in muscle wasting, both directly by increasing energetic inefficiency, and indirectly, by affecting the sarcoplasmic reticulum-mitochondrial assembly that is essential for muscle function and homeostasis. GENERAL SIGNIFICANCE: The present study reports profound morphological changes in cancer cachectic muscle, which are visualized mainly in alterations in sarcoplasmic reticulum and mitochondria. These alterations are linked to pathways that can account for energy inefficiency associated with cancer cachexia.
Asunto(s)
Caquexia/metabolismo , Núcleo Celular/metabolismo , Mitocondrias/metabolismo , Músculo Esquelético/metabolismo , Atrofia Muscular/metabolismo , Sarcoma de Yoshida/metabolismo , Retículo Sarcoplasmático/metabolismo , Translocador 1 del Nucleótido Adenina/genética , Translocador 1 del Nucleótido Adenina/metabolismo , Adenosina Trifosfato/deficiencia , Animales , Apoptosis/genética , Caquexia/complicaciones , Caquexia/patología , Núcleo Celular/ultraestructura , Metabolismo Energético/genética , Expresión Génica , Masculino , Mitocondrias/ultraestructura , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Músculo Esquelético/patología , Atrofia Muscular/complicaciones , Atrofia Muscular/patología , Proteolisis , Ratas , Ratas Wistar , Proteínas Ligasas SKP Cullina F-box/genética , Proteínas Ligasas SKP Cullina F-box/metabolismo , Sarcoma de Yoshida/complicaciones , Sarcoma de Yoshida/patología , Retículo Sarcoplasmático/ultraestructura , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genética , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Proteínas de Motivos Tripartitos , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
OBJECTIVES: Success of selective drug therapies depends on the drug's depot time in the target to treat. Depot time is currently being prolonged, using drug-eluting beads or microspheres for selective internal radiation therapy. The purpose of this study was to establish a model for investigating the depot time of particles injected into tumors in relation to tumor vascularization and particle size. MATERIALS AND METHODS: An animal model with two different vascularized tumors (Walker Carcinoma 256 (hypervascularized) and Yoshida Sarcoma (hypovascularized)) was used. The tumors were implanted into the hind leg of Wistar rats. When the tumors reached 10-15mm, rhenium radiolabeled particles of 25microm and 0.3microm were percutaneously injected into the tumors: large particles (Re-188 microspheres) in 10 hypo- and 10 hypervascularized tumors and small particles (Re-186 sulfid colloid) in 4 hypo- and 16 hypervascularized tumors with the co-injection of the vasoconstrictor, adrenalin (0.01 mg), into 8 hypervascularized tumors. Tumor activity was measured with a gamma camera at 10 min, 1h, 3h, 6h, 14h and 48h p.i. In addition, activity in the lung, liver, spleen, kidneys, and lymph nodes was measured at 48 h p.i. Measurements were adjusted for decay times and then compared. RESULTS: Drainage of the injected particles is bi-phasic, characterized by a fast wash-out. At 10 min p.i., intratumoral activity decreases to 70% of the initially injected activity. This is followed by a slow decline at 48 h p.i in which intratumoral activity decreases to at least 60% of the initially injected activity. Slow decline is independent of particle size and vascularization, whereas fast leakage depends on both. Co-injecting adrenalin significantly reduced the wash-out of the small particles. Radiolabeled microspheres accumulated mainly in the lungs, smaller colloids in the liver. CONCLUSIONS: Particle stay time, biodistribution, and stability can be easily monitored as shown in this animal model. The hematogeneous wash-out can be reduced, using larger particles and vasoconstrictors. Prolonged retention is independent of vascularization and particle size and appears to be sufficient for therapy.
Asunto(s)
Microesferas , Modelos Animales , Neoplasias/metabolismo , Radiofármacos/química , Radiofármacos/farmacocinética , Animales , Carcinoma 256 de Walker/irrigación sanguínea , Carcinoma 256 de Walker/metabolismo , Epinefrina/farmacología , Femenino , Humanos , Inyecciones Intralesiones , Riñón/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Neoplasias/irrigación sanguínea , Radioisótopos/química , Radiofármacos/análisis , Ratas , Ratas Wistar , Renio/química , Sarcoma de Yoshida/irrigación sanguínea , Sarcoma de Yoshida/metabolismo , Albúmina Sérica/química , Bazo/metabolismo , Azufre/química , Distribución Tisular/efectos de los fármacosRESUMEN
Apoptotic events have been clearly associated with muscle wasting in different types of experimental cancer cachexia. In these conditions, cell death is triggered by cytokines or tumour-produced factors. BARD1 is a nuclear protein that is also involved in apoptosis both in vitro and in vivo. The results presented here demonstrate that BARD1 content in skeletal muscle correlates with increased DNA fragmentation during experimental cancer cachexia. It is suggested that BARD1 acts as a modulator of muscle apoptosis or, alternatively, that BARD1 participates in the protein degradation by functioning as ubiquitin ligase.
Asunto(s)
Apoptosis/fisiología , Caquexia/metabolismo , Proteínas Portadoras/biosíntesis , Neoplasias Hepáticas Experimentales/metabolismo , Músculos/patología , Animales , Caquexia/patología , Proteínas Portadoras/genética , Fragmentación del ADN , Inmunohistoquímica , Neoplasias Hepáticas Experimentales/patología , Masculino , Ratas , Ratas Wistar , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/patología , Ubiquitina-Proteína LigasasRESUMEN
In cancer cachexia both cardiac and skeletal muscle suffer an important protein mobilization as a result of increased proteolysis. Administration of the beta2-agonist formoterol to both rats and mice bearing highly cachectic tumors resulted in an important reversal of the muscle-wasting process. The anti-wasting effects of the drug were based on both an activation of the rate of protein synthesis and an inhibition of the rate of muscle proteolysis. Northern blot analysis revealed that formoterol treatment resulted in a decrease in the mRNA content of ubiquitin and proteasome subunits in gastrocnemius muscles; this, together with the decreased proteasome activity observed, suggest that the main anti-proteolytic action of the drug may be based on an inhibition of the ATP-ubiquitin-dependent proteolytic system. Interestingly, the beta2-agonist was also able to diminish the increased rate of muscle apoptosis (measured as DNA laddering as well as caspase-3 activity) present in tumor-bearing animals. The present results indicate that formoterol exerted a selective, powerful protective action on heart and skeletal muscle by antagonizing the enhanced protein degradation that characterizes cancer cachexia, and it could be revealed as a potential therapeutic tool in pathologic states wherein muscle protein hypercatabolism is a critical feature such as cancer cachexia or other wasting diseases.
Asunto(s)
Agonistas Adrenérgicos beta/farmacología , Caquexia/tratamiento farmacológico , Etanolaminas/farmacología , Músculo Esquelético/patología , Neoplasias Experimentales/metabolismo , Animales , Caquexia/metabolismo , Caquexia/patología , Carcinoma Pulmonar de Lewis/metabolismo , Carcinoma Pulmonar de Lewis/patología , Modelos Animales de Enfermedad , Ingestión de Alimentos/efectos de los fármacos , Fumarato de Formoterol , Neoplasias Hepáticas Experimentales/metabolismo , Neoplasias Hepáticas Experimentales/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Musculares/metabolismo , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Neoplasias Experimentales/patología , Ratas , Ratas Wistar , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/patologíaRESUMEN
Induced chemoresistance leads to the reduction of apoptotic responses. Although several drugs are in development that circumvent or decrease existing chemoresistance, none has the potential to prevent or reduce its induction. Here, we present data from a drug that could perhaps fill this gap. Cotreatment of chemotherapy with (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU, RP101) prevented the decrease of apoptotic effects during the course of chemotherapy and reduced nonspecific toxicity. Amplification of chemoresistance genes (Mdr1 and Dhfr) and overexpression of gene products involved in proliferation (DDX1) or DNA repair (UBE2N and APEX) were inhibited, whereas activity of NAD(P)H: quinone oxidoreductase 1 (NQO1) was enhanced. During recovery, when treatment was with BVDU only, microfilamental proteins were up-regulated, and proteins involved in ATP generation or cell survival (STAT3 and JUN-D) were down-regulated. That way, in three different rat tumor models, the antitumor efficiency of chemotherapy was optimized, and toxic side effects were reduced. Because of these beneficial properties of BVDU, a clinical pilot Phase I/II study with five human tumor entities has been started at the University of Dresden (Dresden, Germany). So far, no unwanted side effects have been observed.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Bromodesoxiuridina/análogos & derivados , Bromodesoxiuridina/farmacología , Vinblastina/análogos & derivados , Animales , Bromodesoxiuridina/administración & dosificación , Cisplatino/administración & dosificación , Proteínas de Unión al ADN/metabolismo , Doxorrubicina/administración & dosificación , Resistencia a Antineoplásicos , Sinergismo Farmacológico , Electroforesis en Gel Bidimensional , Fibroblastos/efectos de los fármacos , Ifosfamida/administración & dosificación , Metotrexato/administración & dosificación , Metotrexato/farmacología , Ratones , Mitomicina/administración & dosificación , Mitoxantrona/administración & dosificación , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Transcripción STAT3 , Sarcoma de Yoshida/tratamiento farmacológico , Sarcoma de Yoshida/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Transactivadores/metabolismo , Células Tumorales Cultivadas , Vinblastina/administración & dosificación , VinorelbinaRESUMEN
The immune response is modulated by genetic adjuvants using plasmid vectors expressing cytokines. Skeletal muscle can express a foreign gene intramuscularly administered via a needle injection, and the potential of muscle as a target tissue for somatic gene therapy in treating cancer has been explored. In the present study, we investigated the efficacy of particle-mediated intramuscular transfection modified with a local anesthetic agent, bupivacaine, on luciferase and green fluorescent protein. The results indicate that these proteins are more efficiently expressed and persist longer in muscle modified in this way compared with the needle-injection method. Using an established rat sarcoma model, particle-mediated intramuscular gene-gun therapy with a combination of IL-12 and IL-18 cDNA was conducted. Growth of the distant sarcoma was significantly inhibited by particle-mediated intramuscular combination gene therapy, and the survival rate was also improved. Furthermore, the combination gene-gun therapy maintained significant levels of interferon-gamma and induced a high activity of tumor-specific cytotoxic T lymphocytes. These results suggest that the sustained local delivery of IL-12 and IL-18 cDNA using intramuscular gene-gun therapy modified with bupivacaine can induce long-term antitumor immunity, and can provide the great advantage of inhibiting the disseminated tumor.
Asunto(s)
Anestésicos Locales/uso terapéutico , Biolística , Bupivacaína/uso terapéutico , Terapia Genética/métodos , Interleucina-12/genética , Interleucina-18/genética , Sarcoma de Yoshida/terapia , Animales , Bupivacaína/farmacología , ADN Complementario , Expresión Génica , Inyecciones Intramusculares , Interferón gamma/sangre , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Interleucina-18/metabolismo , Cinética , Masculino , Músculo Esquelético/metabolismo , Plásmidos/administración & dosificación , Ratas , Ratas Endogámicas Lew , Sarcoma de Yoshida/inmunología , Sarcoma de Yoshida/metabolismo , Células TH1/inmunología , TransfecciónRESUMEN
Implantation of Yoshida ascites sarcoma in rats was found to lead to a reduction in the hemoglobin content, the erythrocyte count and the packed cell volume of blood to 30% of normal in 4 d; however, there was no decrease in the mean cell hemoglobin, the mean cell volume and the mean corpuscular hemoglobin concentration, or suppression of erythropoiesis. The red cells from the circulation of tumor-bearing animals, tagged with (51)Cr and injected intravenously in normal rats, showed significantly faster clearance than normal. The erythrocytes contaminating the tumor ascites exhibited extremely short survival, suggesting that one or more secreted tumor product(s) may be responsible for the effect. Incubation of red cells from normal rats in the cell-free ascites fluid, or with an isoform of alpha2-macroglobulin purified from it, also led to reduction in the survival; but the ascites fluid depleted specifically of alpha2-macroglobulin was without any effect. The erythrocytes exhibiting reduced survival showed a proportionate decrease in their cellular deformability. The study identifies a tumor product that is directly responsible for the causation of anemia in the host, and the mechanism by which it does so.
Asunto(s)
Anemia/etiología , Envejecimiento Eritrocítico , Deformación Eritrocítica , Eritrocitos Anormales/patología , Proteínas de Neoplasias/fisiología , Síndromes Paraneoplásicos/etiología , Sarcoma de Yoshida/sangre , alfa-Macroglobulinas/fisiología , Anemia/sangre , Animales , Proteína 1 de Intercambio de Anión de Eritrocito/análisis , Líquido Ascítico/química , Agregación Eritrocitaria , Síndromes Paraneoplásicos/sangre , Ratas , Ratas Wistar , Sarcoma de Yoshida/complicaciones , Sarcoma de Yoshida/metabolismoRESUMEN
The development of new pharmacological approaches for preventing muscle wasting in cancer is an important goal because cachectic patients display a reduced response to chemotherapy and radiotherapy. Xanthine derivatives such as pentoxifylline inhibit tumour necrosis factor-alpha (TNF) production, which has been implicated in the signalling of muscle wasting. However, the effect of pentoxifylline has been inconclusive in clinical trials. We report here the first direct evidence that daily injections of torbafylline (also known as HWA 448), another xanthine derivative, had no effect by itself on muscle proteolysis in control healthy rats. In cancer rats, the drug blocked the lipopolysaccharide-induced hyperproduction of TNF and prevented muscle wasting. In these animals HWA 448 suppressed the enhanced proteasome-dependent proteolysis, which is sensitive to the proteasome inhibitor MG132, and the accumulation of high-molecular-mass ubiquitin (Ub) conjugates in the myofibrillar fraction. The drug also normalized the enhanced muscle expression of Ub, which prevails in the atrophying muscles from cancer rats. In contrast, HWA 448 did not reduce the increased expression of either the 14 kDa Ub conjugating enzyme E2 or the ATPase and non-ATPase subunits of the 19 S regulatory complex of the 26 S proteasome, including the non-ATPase subunit S5a, which recognizes polyUb degradation signals. Finally, the drug also prevented muscle wasting in septic rats (which exhibit increased TNF production), and was much more potent than pentoxifylline or other xanthine derivatives. Taken together, the data indicate that HWA 448 is a powerful inhibitor of muscle wasting that blocks enhanced Ub-proteasome-dependent proteolysis in situations where TNF production rises, including cancer and sepsis.
Asunto(s)
Cisteína Endopeptidasas/metabolismo , Complejos Multienzimáticos/metabolismo , Músculo Esquelético/efectos de los fármacos , Pentoxifilina/análogos & derivados , Pentoxifilina/farmacología , Sarcoma de Yoshida/metabolismo , Sepsis/metabolismo , Ubiquitina/metabolismo , Animales , Cisteína Endopeptidasas/genética , Regulación hacia Abajo/efectos de los fármacos , Hidrólisis , Masculino , Complejos Multienzimáticos/genética , Músculo Esquelético/enzimología , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Complejo de la Endopetidasa Proteasomal , ARN Mensajero/genética , Ratas , Ratas Wistar , Sarcoma de Yoshida/enzimología , Sepsis/enzimología , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
It is known that cisplatin (CDDP) potentiates the cytotoxicity of 5-fluorouracil (5-FU), and that the biochemical mechanism is an increase in the intracellular reduced folate levels in the tumor cells. We investigated the effect of consecutive administration with lower-dose CDDP on intracellular accumulation of reduced folate and the activity of methionine synthase, a key enzyme in intracellular methionine synthesis. When CDDP (1 mg/kg) was administered i.p. to ascitic Yoshida sarcoma-bearing rats for 4 consecutive days, both the reduced folate levels and methionine synthase activity in the cells significantly increased, as the same as a single 5 mg/kg dose of CDDP. Furthermore, when Yoshida sarcoma-bearing rats were pre-treated with 1 mg/kg CDDP for 5 consecutive days, [14C]L-methionine incorporation into the isolated ascitic cells was significantly inhibited as compared to that in non-treated cells, suggesting that consecutive administration of lower-dose CDDP is capable of inducing the intracellular modulation of reduced folate levels and methionine synthase activity via inhibition of cellular uptake of methionine. In addition, 5-day administration of lower-dose (1 mg/kg) CDDP potentiated the antitumor effect of 5 mg/kg S-1, a new oral preparation of tegafur, given for 7 consecutive days, and this combined effect was almost similar to the antitumor effect of a combination of S-1 and a single conventional dose (5 mg/kg) of CDDP. Consecutive lower-dose CDDP also may be concluded to act as an important modulator of the enhancement of 5-FU cytotoxicity in experimental tumors.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/toxicidad , Cisplatino/farmacología , Fluorouracilo/toxicidad , Sarcoma de Yoshida/tratamiento farmacológico , 5-Metiltetrahidrofolato-Homocisteína S-Metiltransferasa/biosíntesis , 5-Metiltetrahidrofolato-Homocisteína S-Metiltransferasa/metabolismo , Animales , Cisplatino/administración & dosificación , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Combinación de Medicamentos , Sinergismo Farmacológico , Inducción Enzimática/efectos de los fármacos , Fluorouracilo/administración & dosificación , Ácido Fólico/metabolismo , Masculino , Metionina/farmacocinética , Ratones , Trasplante de Neoplasias , Ácido Oxónico/administración & dosificación , Piridinas/administración & dosificación , Ratas , Sarcoma de Yoshida/metabolismo , Tegafur/administración & dosificaciónRESUMEN
S-1, a new oral 5-fluorouracil (5-FU)-derivative antitumor agent, is composed of tegafur, 5-chloro-2,4-dihydropyridine, and potassium oxonate (Oxo). Oxo, which inhibits the phosphorylation of 5-FU, is added to reduce the gastrointestinal (GI) toxicity of the agent. In this study, we investigated the tissue distribution and the metabolic fate of Oxo in rats after oral administration of S-1. Oxo was mainly distributed to the intracellular sites of the small intestines in a much higher concentration than 5-FU, but little distributed to other tissues, including tumorous ones in which 5-FU was observed after oral administration of S-1. Plasma concentration-time profiles of Oxo and its metabolites after i.v. and oral administration of S-1 revealed that Oxo was mainly converted to cyanuric acid in the GI tract. Furthermore, the analysis of drug-related radioactivity in GI contents and in vitro studies suggested that Oxo was converted to cyanuric acid by two routes, the first being direct conversion by the gut flora in the cecum, and the second, conversion by xanthine oxidase or perhaps by aldehyde oxidase after degradation to 5-azauracil (5-AZU) by the gastric acid. These results indicate that, although a part of the administered Oxo was degraded in the GI tract, Oxo was mainly distributed to the intracellular sites of the small intestines in a much higher concentration than 5-FU and that little was distributed to other tissues, including tumors. We conclude that this is the reason why Oxo suppresses the GI toxicity of 5-FU without affecting its antitumor activity.
Asunto(s)
Antimetabolitos Antineoplásicos/farmacocinética , Ácido Oxónico/farmacocinética , Piridinas/farmacocinética , Tegafur/farmacocinética , Uracilo/análogos & derivados , Administración Oral , Alopurinol/farmacología , Animales , Antimetabolitos Antineoplásicos/metabolismo , Área Bajo la Curva , Biotransformación , Radioisótopos de Carbono , Clorpromazina/farmacología , Combinación de Medicamentos , Medicamentos Herbarios Chinos/farmacología , Fluorouracilo/sangre , Fluorouracilo/metabolismo , Glycyrrhiza , Intestino Delgado/metabolismo , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Ácido Oxónico/sangre , Ácido Oxónico/metabolismo , Paeonia , Piridinas/sangre , Piridinas/metabolismo , Ratas , Ratas Endogámicas , Sarcoma de Yoshida/tratamiento farmacológico , Sarcoma de Yoshida/metabolismo , Tegafur/sangre , Tegafur/metabolismo , Distribución Tisular , Triazinas/sangre , Triazinas/metabolismo , Uracilo/metabolismo , Xantina Oxidasa/metabolismoRESUMEN
The results presented herein clearly indicate that nitroxide derivatives--free radicals are effective as substrates for one-electron oxidation in the peroxidase cycle involving hydrogen peroxide, which have been the subject of considerable controversy. This oxidation is catalyzed enzymatically and it might occur in tumor cells (in vivo) where the level of ROS (H2O2 and O2.-) is increased. The result of this reaction involving hydrogen peroxide is the obligative formation of the oxo-ammonium cation involved in the superoxide dismutase-mimic reaction of nitroxides with superoxide and/or in reaction with H2O2 leading to superoxide formation and regeneration of the parent nitroxide molecule. The efficiency of this enzymatically catalyzed oxidation of nitroxide(s) depends on the structure of the substituent in position 4 of nitroxide ring as follows: -OCH3 > -NHCOCH3 > -NHCOCH2CH3. Notably, the reduced nitroxide salt was not substrate for peroxidatic oxidation clearly indicating the importance of the free radical moiety of the nitroxide molecule. These findings may have some relevance in the recent investigations of antioxidant properties/mechanisms of nitroxides. Based on these considerations we hypothesize that the administration of oxidizable free radical nitroxide compounds--antioxidants may be a useful strategy in the treatment and investigations of cancer diseases. An in vivo study ("Screening test of chemicals employing Yoshida Sarcoma animals") was carried out to verify whether the structure and/or the chain length of substituent of oxidizable nitroxide derivatives--antioxidants could influence their apoptotic activity. The results reported in this study are encouraging as we found a limited correlation between the molecular oxidative properties of nitroxides under study, their structure and antitumor (apoptotic) action. In conclusion, this work demonstrates that investigation of the structure-dependent oxidation of antioxidatively acting nitroxides can become a very important step in their future screening and selection for applications in vivo and in vitro.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Piperidinas/farmacología , Sarcoma de Yoshida/tratamiento farmacológico , Animales , Femenino , Oxidación-Reducción , Ratas , Ratas Wistar , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/patología , Relación Estructura-ActividadRESUMEN
Here we show for the first time that the model nitroxide derivatives, free radical or its reduced piperidinium salt, suppressed cytotoxicity of ROS (O2 and H2O2) generated outside the cells (B14 line, model for neoplastic phenotype) in ***. The nitroxides prevented the decrease in the number of *** caused by exogenous O2- and H2O2 at concentrations which were not themselves cytotoxic. In the present study, we have also shown that a very substantial difference in the cell response occurred when the model rat tumor cells (Yoshida Sarcoma ascites) were treated in vivo with six novel synthesized nitroxide antioxidants. A number of tumor cells displayed morphological characteristics of apoptosis. This effect was comparable to those observed for other nitroxyls under similar experimental conditions. Since the increase in the ROS generation followed by apoptotic changes of nuclei is the consistent recent finding in various experimental models of apoptosis, one fundamental question was raised: why nitroxide antioxidants paradoxically act as apoptosis inducers in vivo? Taking together the results presented here and in our previous works, it seems reasonable to suggest that nitroxide-antioxidants improve the endogenous "antioxidants reserve" and action can induce a reductive stress as opposed to an oxidative stress, triggering a cascade of dose-dependent processes involving indirectly an antioxidant mechanism(s) and resulting in the apoptotic death of cancer cells in vivo. The SAR (structure activity relationship) revealed that either the substituent structure at 4-position of the nitroxide ring or its oxidation state are determinant for the degree of the observed differences in the apoptotic potency of nitroxide derivates in vivo.
Asunto(s)
Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Sarcoma de Yoshida/tratamiento farmacológico , Animales , Cricetinae , Relación Dosis-Respuesta a Droga , Masculino , Ratas , Especies Reactivas de Oxígeno/metabolismo , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/patología , Relación Estructura-ActividadRESUMEN
Tissue protein hypercatabolism (TPH) is an important feature in cancer cachexia, particularly with regard to the skeletal muscle. The Yoshida AH-130 rat ascites hepatoma is a model system for studying the mechanisms involved in the processes that lead to tissue depletion, since it induces in the host a rapid and progressive muscle wasting, primarily due to TPH. The present study was aimed at investigating if IL-15, which is known to favour muscle fibre hypertrophy, could antagonize the enhanced muscle protein breakdown in this cancer cachexia model. Indeed, IL-15 treatment partly inhibited skeletal muscle wasting in AH-130-bearing rats by decreasing (8-fold) protein degradative rates (as measured by 14C-bicarbonate pre-loading of muscle proteins) to values even lower than those observed in non-tumour-bearing animals. These alterations in protein breakdown rates were associated with an inhibition of the ATP-ubiquitin-dependent proteolytic pathway (35% and 41% for 2.4 and 1.2 kb ubiquitin mRNA, and 57% for the C8 proteasome subunit, respectively). The cytokine did not modify the plasma levels of corticosterone and insulin in the tumour hosts. The present data give new insights into the mechanisms by which IL-15 exerts its preventive effect on muscle protein wasting and seem to warrant the implementation of experimental protocols involving the use of the cytokine in the treatment of pathological states characterized by TPH, particularly in skeletal muscle, such as in the present model of cancer cachexia.
Asunto(s)
Caquexia/tratamiento farmacológico , Interleucina-15/farmacología , Neoplasias Hepáticas Experimentales/metabolismo , Proteínas Musculares/metabolismo , Músculo Esquelético/efectos de los fármacos , Sarcoma de Yoshida/metabolismo , Animales , Caquexia/etiología , Caquexia/metabolismo , Neoplasias Hepáticas Experimentales/complicaciones , Masculino , Músculo Esquelético/metabolismo , Atrofia Muscular/tratamiento farmacológico , Atrofia Muscular/etiología , Atrofia Muscular/metabolismo , Trasplante de Neoplasias , Ratas , Ratas Wistar , Sarcoma de Yoshida/complicacionesRESUMEN
The development of pharmacological approaches for preventing the loss of muscle proteins would be extremely valuable for cachectic patients. For example, severe wasting in cancer patients correlates with a reduced efficacy of chemotherapy and radiotherapy. Pentoxifylline (PTX) is a very inexpensive xanthine derivative, which is widely used in humans as a haemorheological agent, and inhibits tumor necrosis factor transcription. We have shown here that a daily administration of PTX prevents muscle atrophy and suppresses increased protein breakdown in Yoshida sarcoma-bearing rats by inhibiting the activation of a nonlysosomal, Ca(2+)-independent proteolytic pathway. PTX blocked the ubiquitin pathway, apparently by suppressing the enhanced expression of ubiquitin, the 14-kDa ubiquitin conjugating enzyme E2, and the C2 20S proteasome subunit in muscle from cancer rats. The 19S complex and 11S regulator associate with the 20S proteasome and regulate its peptidase activities. The mRNA levels for the ATPase subunit MSS1 of the 19S complex increased in cancer cachexia, in contrast with mRNAs of other regulatory subunits. This adaptation was suppressed by PTX, suggesting that the drug inhibited the activation of the 26S proteasome. This is the first demonstration of a pharmacological manipulation of the ubiquitin-proteasome pathway in cachexia with a drug which is well tolerated in humans. Overall, the data suggest that PTX can prevent muscle wasting in situations where tumor necrosis factor production rises, including cancer, sepsis, AIDS and trauma.
Asunto(s)
Caquexia/metabolismo , Cisteína Endopeptidasas/metabolismo , Complejos Multienzimáticos/metabolismo , Músculo Esquelético/enzimología , Pentoxifilina/farmacología , Péptido Hidrolasas/metabolismo , Sarcoma de Yoshida/metabolismo , Adenosina Trifosfatasas/metabolismo , Animales , Caquexia/etiología , Activación Enzimática/efectos de los fármacos , Proteínas Musculares/metabolismo , Músculo Esquelético/efectos de los fármacos , Inhibidores de Fosfodiesterasa/farmacología , Complejo de la Endopetidasa Proteasomal , Ratas , Sarcoma de Yoshida/fisiopatologíaRESUMEN
This study was undertaken to elucidate the relationship between the biodistribution of radioactive metal nuclides in tumor tissue and its physicochemical properties. Potassium analogs (86Rb, 134Cs, 201Tl) were taken up into viable tumor tissue, although 22Na concentrated in necrotic tumor tissue. 67Ga and 111In were more predominant in inflammatory tissue than in the viable and necrotic tumor tissue. 169Yb and 167Tm accumulated in viable tumor tissue and tissue containing viable and necrotic tumor tissue. 67Ga, 111In, 169Yb and 167Tm were bound to the acid mucopolysaccharide with a mol. wt. of about 10,000 daltons in the tumor tissue. 46Sc, 51Cr, 95Zr, 181Hf, 95Nb, 182Ta, and 103Ru were highly concentrated in inflammatory tissue and were bound to the acid mucopolysaccharides with a mol. wt. exceeding 40,000 daltons. 65Zn and 103Pd concentrated in viable tumor tissue and were bound to the protein in the tissue. The results suggest that the difference in intra-tumor distribution of these elements is caused by a difference in the binding substances (or status) of these elements in the tissues, and the binding substance is determined by physicochemical properties of the elements. We therefore conclude that the biodistribution of radioactive metal ions in tumor tissue is determined by its own physicochemical properties.
Asunto(s)
Carcinoma de Ehrlich/metabolismo , Metales/farmacocinética , Radioisótopos/farmacocinética , Sarcoma de Yoshida/metabolismo , Animales , Carcinoma de Ehrlich/diagnóstico por imagen , Tejido Conectivo/metabolismo , Glicosaminoglicanos/metabolismo , Inflamación , Masculino , Necrosis , Unión Proteica , Cintigrafía , Ratas , Sarcoma de Yoshida/diagnóstico por imagen , Distribución TisularRESUMEN
S-1 is an oral combined form of 1 M tegafur [a prodrug of 5-fluorouracil (5-FU)], 0.4 M 5-chloro-2,4-dihydroxypyridine (a reversible inhibitor of dihydropyrimidine dehydrogenase) and 1 M potassium oxonate (an inhibitor of orotate phosphoribosyltransferase). S-1 has been shown to exert a potent antitumor effect with low gastrointestinal toxicity in experimental tumor models. We have therefore compared the antitumor effect of oral S-1 with that of continuous infusion of 5-FU in rats bearing transplants of human and murine tumors. Almost complete inhibition of the tumor growth was obtained on 7 day schedules in Yoshida sarcoma-bearing rats by consecutive administration of 30 mg/kg/day of oral S-1 and 40 mg/kg/day infusion of 5-FU. However, a significant difference between the incidence of toxicities of S-1 and 5-FU, including body weight loss and diarrhea, was noted. The rats given the 5-FU infusion had marked weight loss and severe diarrhea, while those given oral S-1 had neither. Although about 50% inhibition of the tumor growth was attained with 15 mg/kg/day of oral S-1 and 30 mg/kg/day infusion of 5-FU in nude rats with xenografted human colon cancer (KM12C), the rate of body weight loss in the 5-FU-treated group was distinctly higher than in the S-1-treated group. The ratio of the 5-fluoronucleotide concentrations in gastrointestinal tissue to that in the tumor was lower in the S-1-treated rats than in the 5-FU-treated rats. In conclusion, the results suggest that oral S-1 might be more effective in the treatment of cancer patients than continuous infusion of 5-FU, from the standpoint of antitumor potency and toxicity.
Asunto(s)
Antimetabolitos Antineoplásicos/farmacología , Fluorouracilo/farmacología , Ácido Oxónico/farmacología , Piridinas/farmacología , Tegafur/farmacología , Administración Oral , Animales , Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/efectos adversos , División Celular/efectos de los fármacos , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/metabolismo , Sistema Digestivo/metabolismo , Combinación de Medicamentos , Fluorouracilo/efectos adversos , Fluorouracilo/metabolismo , Humanos , Infusiones Intravenosas , Trasplante de Neoplasias , Ácido Oxónico/administración & dosificación , Ácido Oxónico/efectos adversos , Piridinas/administración & dosificación , Piridinas/efectos adversos , Ratas , Ratas Endogámicas F344 , Sarcoma de Yoshida/tratamiento farmacológico , Sarcoma de Yoshida/metabolismo , Tegafur/administración & dosificación , Tegafur/efectos adversos , Trasplante HeterólogoRESUMEN
Boron neutron capture therapy (BNCT) may improve the locoregional control of radio/chemoresistant tumours like soft tissues sarcomas (STS). This technique uses the 10B(n,alpha)7Li nuclear reaction to destroy tumour cells, provided that a sufficient amount of 10B may be carried selectively into them. In order to evaluate the targeting potential of 10B-L-borophenylalanine (BPA) a 10B biodistribution study was carried out in 24 Wistar rats bearing Yoshida sarcoma. Six animals received increasing intraperitoneal doses of BPA (300, 600 and 1200 mg kg-1), while the remainder received a BPA dose of 600 mg kg-1 but with a sacrifice at six different time points: 1, 2, 4, 6, 9 and 12 h. The 10B concentrations in the tumours, normal tissues and blood were analysed with neutron capture radiography (NCR). The analysis shows that 36 micrograms g-1 (+/- 4 SD) of 10B may be incorporated into the tumour, with a ratio of 13 (+/- 4 SD) versus the muscle and a ratio of 15 (+/- 3 SD) versus the blood, 6 h after an intraperitoneal injection of 600 mg kg-1 of BPA. The BPA appears to be abundantly incorporated in the tumour, and the kidney proximal tubule area. These data suggest that BNCT using BPA may provide an improved therapeutic ratio for the treatment of STS.
Asunto(s)
Compuestos de Boro/análisis , Compuestos de Boro/farmacocinética , Terapia por Captura de Neutrón de Boro , Sarcoma de Yoshida/metabolismo , Animales , Trasplante de Neoplasias , Ratas , Ratas Wistar , Sarcoma de Yoshida/radioterapiaRESUMEN
Microspheres containing cisplatin (CDDP) embedded in poly-d,l-lactic acid (PLA) and polyethylene glycol acid (CDDP-PPMS) were developed to improve treatment of malignant effusions. In vitro studies demonstrated that CDDP was released continuously for more than 4 weeks from CDDP-PPMS without initial burst. CDDP-PPMS was compared with CDDP aqueous solution (CDDP-SOL) by i.p. administration in rats for 1) tissue distribution, 2) toxicity and 3) therapeutic effects against Yoshida sarcoma. We found that the CDDP concentration in the omentum was maintained at a higher level than in the CDDP-SOL group, while the particles of CDDP-PPMS were observed in the stomata of the omentum by electron microscopy. Concentrations of CDDP in the lung, liver, kidney and blood were lower in the CDDP-PPMS group than in the CDDP-SOL group. All rats given CDDP-PPMS containing < or = 28 mg/kg were alive, whereas in the CDDP-SOL group, all rats given > or = 16 mg/kg died from side effects. The LD50 of CDDP-PPMS and CDDP-SOL were 32.8 and 14.8 mg/kg, respectively. The survival of rats with peritoneal metastasis was better in the CDDP-PPMS group than in the CDDP-SOL group.
Asunto(s)
Cisplatino/administración & dosificación , Ácido Láctico/administración & dosificación , Neoplasias Peritoneales/tratamiento farmacológico , Vehículos Farmacéuticos/administración & dosificación , Polietilenglicoles/administración & dosificación , Polímeros/administración & dosificación , Sarcoma de Yoshida/tratamiento farmacológico , Animales , Química Farmacéutica , Cisplatino/farmacocinética , Cisplatino/toxicidad , Inyecciones Intraperitoneales , Ácido Láctico/química , Ácido Láctico/farmacocinética , Masculino , Microesferas , Trasplante de Neoplasias , Neoplasias Peritoneales/metabolismo , Neoplasias Peritoneales/secundario , Vehículos Farmacéuticos/química , Vehículos Farmacéuticos/farmacocinética , Poliésteres , Polietilenglicoles/química , Polietilenglicoles/farmacocinética , Polímeros/química , Polímeros/farmacocinética , Ratas , Sarcoma de Yoshida/metabolismo , Sarcoma de Yoshida/secundario , Distribución TisularRESUMEN
Arginine supplementation increases glutamine levels in muscle and plasma. Since glutamine production is increased in catabolic states, these observations prompted us to investigate whether the flux of arginine to glutamine was increased in tumor-bearing (TB) rats, and we measured the synthesis rate of glutamine from arginine in control versus TB rats receiving standard total parenteral nutrition (TPN) solution. Male Donryu rats (N = 36; body weight, 200 to 225 g) were divided into two groups, control and TB rats. Yoshida sarcoma cells (1 x 10(6)) were inoculated into the back of the rats (n = 18) subcutaneously on day 0. The rats were given free access to water and rat chow. On day 5, all animals, including non-TB rats (n = 18), were catheterized at the jugular vein and TPN was begun. On day 10, TPN solution containing either U-14C-glutamine (2.0 microCi/h) or U-14C-arginine (2.0 microCi/h) was infused as a 6-hour constant infusion. At the end of the isotope infusion, plasma was collected to determine the glutamine production rate in rats receiving U-14C-glutamine, and the ratio of specific activity of glutamine to specific activity of arginine was measured in rats receiving U-14C-arginine. Only 2 g tumor caused a decrease in glutamine levels and an increase in glutamine and arginine production. The low flux rate of arginine to glutamine was observed in control rats (Arg to Gln, 41.0 +/- 11.9 mumol/kg/h). On the other hand, TB caused a significant increase in Arg to Gln compared with the control (213.3 +/- 66.1 mumol/kg/h, P < .01 v control). An increase in the flux rate of Arg to Gln was associated with an enhancement in the ratio of specific activity of ornithine to specific activity of arginine in TB rats (control 51.5% +/- 10.9% v 77.4% +/- 8.9%, P < .05). We conclude that (1) glutamine and arginine metabolism is altered with very small tumors, (2) although the flux of Arg to Gln was increased in TB and rats, the small increase in Arg to Gln cannot explain the observed large increase in Gln production.