RESUMEN
AIM: The aim of this study was to determine the effect of morphine on bladder cancer cell proliferation and apoptosis in vitro. MATERIALS AND METHODS: MTT assay was used to measure percentage growth of RT-112 human bladder cancer cells after 72 hours of morphine/morphine + naloxone treatment. Expression of µ-opioid receptors was assessed by Western blot and finally, apoptotic assay with CellEvent Caspase-3/7 Green Detection Reagent was carried out using confocal microscopy. RESULTS: The MTT assays showed that morphine increased RT-112 cell growth. Naloxone inhibited this growth enhancing effect. Western blot analysis regarding µ-opioid receptor expression in RT-112 cells remains inconclusive. Morphine was also found to decrease the rate of apoptosis of RT-112 cells, an effect which naloxone inhibited. CONCLUSIONS: This study provides evidence that morphine, at clinically relevant doses, causes RT-112 bladder cancer cell proliferation, possibly opioid receptor mediated and at least some of this effect might be due to decreased apoptosis. Clinically, this suggests that in patients with bladder cancer, managing pain with morphine might have detrimental consequences on patient outcomes and alternative pain relief should be considered if possible.
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Proliferación Celular/efectos de los fármacos , Morfina/farmacología , Receptores Opioides mu/genética , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Apoptosis/efectos de los fármacos , Caspasas/genética , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Naloxona/farmacología , Neoplasias de la Vejiga Urinaria/metabolismo , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
OBJECTIVE: To investigate possible effects of honey on angiogenesis, using in vitro analogues of angiogenesis and an endothelial proliferation assay. METHOD: Using an in vitro rat aortic ring assay we compared pseudotubule formation by medicinal honey (Activon), supermarket honey (Rowse) and a honey-based ointment (Mesitran), with that of artificial honey (70% w/w sugar glucose/fructose). Pseudotubules were analysed using TCS Cellworks AngioSys software. The Angiokit sytem was used to validate the results. Using the MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium. Bromide] assay, toxicity was also assessed on human umbilical vein endothelial cells (HUVEC) directly adherent to plastic. RESULTS: All honey preparations stimulated pseudotubule formation, maximal at around 0.2% honey. Medicinal honeys were more active than Rowse. The effect was not attributable to the sugar content. Among the honeys tested, the Manuka-based Activon preparation reduced residual viable biomass compared with a sugar control at > 0.32% v/v concentration. Rowse had a similar effect only at 2.5%, the highest dose tested. CONCLUSION: The influence of honey constituents on angiogenesis in a wound dressing context is likely to be positive, but would depend on the effective dilution of the honey and the penetration of the active constituents against an osmotic gradient. The extent to which this occurs has yet to be established. CONFLICT OF INTEREST: This work was conceived, designed and executed by the authors. Medical honey preparations were supplied unconditionally but free of charge by the distributors.
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Miel , Neovascularización Fisiológica/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Heridas y Lesiones/terapia , Administración Cutánea , Animales , Aorta/citología , Biomasa , Pruebas Inmunológicas de Citotoxicidad , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Miel/normas , Miel/provisión & distribución , Humanos , Pomadas , Ósmosis , Ratas , Cuidados de la Piel/métodos , Sales de Tetrazolio , Tiazoles , Venas Umbilicales/citologíaRESUMEN
OBJECTIVE: To determine if Meglumine-Eicosapentaenoic Acid (MeEPA) acts synergistically with epirubicin and mitomycin to enhance cytotoxicity towards bladder cancer cell lines in vitro. MATERIALS AND METHODS: Bladder cancer cells were exposed to MeEPA in combination with epirubicin or mitomycin. Residual viable cell biomass was estimated with the methyl-thiazoldiphenyl tetrazolium (MTT) assay following drug exposure. Drug interaction was analysed using median effect analysis to determine levels of synergism. RESULTS: Most combinations of MeEPA with both epirubicin and mitomycin showed a high-level of synergism. At high doses, drug precipitation adversely affected MTT assay analysis suggesting antagonism of action. However, the predominant pattern was of synergism for most dose combinations tested. CONCLUSION: Bladder cancer treated by endoscopic resection alone is subject to high recurrence rates. Post-operative intravesical instillation of epirubicin and mitomycin can halve recurrence rates, but there is no evidence that disease progression to invasive bladder cancer is altered. Thus, optimisation of current treatment strategies is required. The anti-tumour activity of fatty acids is well established and MeEPA is a new, soluble formulation with the potential to enhance intravesical drug efficacy.
Asunto(s)
Antibióticos Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica , Ácido Eicosapentaenoico/uso terapéutico , Epirrubicina/uso terapéutico , Meglumina/uso terapéutico , Mitomicina/uso terapéutico , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Línea Celular Tumoral , Combinación de Medicamentos , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Ácidos Grasos Omega-3/uso terapéutico , HumanosRESUMEN
Prostasomes are organelles secreted by prostatic epithelial cells, and are believed to have a role in fertility and prostatic disease. They are known to influence sperm motility and the acrosome reaction, and are thought to have a role in cell transformation, immunosuppression, proliferation, facilitation of local invasion, and angiogenesis. Previously, we have demonstrated the inhibitory effect of prostasomes derived from human semen on angiogenesis using HUVEC cells grown on matrigel. In this study, we use the rat aortic ring assay system, arguably a closer reflection of the in vivo situation. Quantification was by a spectrophotometric method, and underlying mechanisms assessed. Prostasomes demonstrated a clear inhibition of angiogenesis, and this effect persisted after heat treatment of prostasomes to denature protein. This fits with other known effects of prostasomes known to be due to the membrane lipid component, which is unusually high in sphingomyelin and cholesterol.
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Aorta Torácica/patología , Células Epiteliales/metabolismo , Neovascularización Patológica/prevención & control , Próstata/citología , Vesículas Seminales/citología , Animales , Técnicas de Cultivo de Célula , Humanos , Masculino , Orgánulos , Ratas , Ratas Wistar , Proyectos de Investigación , Vesículas Seminales/metabolismo , EspectrofotometríaRESUMEN
BACKGROUND: Multidrug resistance (MDR) has a potentially serious influence on cancer treatment and should be taken into consideration in the design and application of therapeutic regimens. It is mediated through the activity of cellular pumps. AIM: To investigate whether furosemide, itself a pump-blocker, reverses MDR in an in vitro model. MATERIALS AND METHODS: An MDR bladder cancer cell line (MGH-u 1R) and its parental (drug sensitive) clone were exposed to epirubicin and furosemide, with the concentration of one drug fixed and that of the other serially diluted in a 96-well plate format. Both drugs formed the variable component in separate experiments. After a 1-h exposure, the cells were washed and replenished with fresh medium. To examine the toxicity of epirubicin and furosemide separately and in combination, monotetrazolium-based assays were carried out. Intracellular epirubicin distribution was assessed by confocal microscopy as a second index of resistance status after in vitro exposure. RESULTS: MGH-u 1R cells incubated with furosemide showed distribution of drug similar to that in the parental cells (MGH-u 1 sensitive). Controls (without furosemide) continued to show a resistant pattern of fluorescence. In cytotoxicity assays furosemide appeared substantially non-toxic. Resistant cells in the toxicity titration experiments showed increased resistance to levels of furosemide over 500 mug/ml. Parental cells were made only marginally more sensitive against increased background toxicity. CONCLUSION: Furosemide is effective in reversing MDR status in bladder cancer cell lines in vitro. It may also have an increment of intrinsic cytotoxicity, but only at higher concentrations. We propose a potential for further investigation of furosemide as an adjunct to chemotherapy for superficial bladder cancer.
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Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Furosemida/farmacología , Inhibidores del Simportador de Cloruro Sódico y Cloruro Potásico/farmacología , Neoplasias de la Vejiga Urinaria/patología , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/farmacología , Muerte Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Epirrubicina/farmacocinética , Epirrubicina/farmacología , Humanos , Microscopía Confocal , Células Tumorales CultivadasRESUMEN
Lycopene (C(40) H(56)) is a highly lipophilic antioxidant found in human semen in nanomolar concentrations. It has been shown to be one of the most potent carotenoid antioxidant in various human studies. Prostasomes are organelles secreted by glandular prostatic epithelial cells and are known to play an important role in fertility and prostate cancer. They are also known to possess antioxidant activity and aid the functioning of sperm. We studied the ability of these vesicles to adsorb and retain lycopene into their rich lipid environment in vitro. High-performance liquid chromatography analysis confirmed micrograms of lycopene per milligram of prostasomal protein. In view of the prostasomes' lipid-rich nature it is highly likely that these organelles act as delivery vehicles for this highly lipophilic antioxidant substance into human semen.
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Antioxidantes/farmacocinética , Carotenoides/farmacocinética , Orgánulos/metabolismo , Próstata/metabolismo , Semen/metabolismo , Adsorción , Carotenoides/química , Cromatografía Líquida de Alta Presión , Células Epiteliales/metabolismo , Humanos , Técnicas In Vitro , Licopeno , Masculino , Próstata/ultraestructura , SolucionesRESUMEN
Prostasomes are biologically active organelles that are secreted by human prostate epithelial cells, and it is believed that they have a role in prostatic disease. We studied the effect of prostasomes on the human umbilical vein endothelial cell (HUVEC)/Matrigel model of angiogenesis, and the association of labelled prostasomes with HUVECs. The growth inhibitory effect of prostasomes on HUVECs was assayed by spectrophotometric measurement of residual biomass. Preparations of HUVECs on a Matrigel base were exposed to prostasomes, and the development of capillary-like networks was quantified. Prostasomes were labelled with PKH-26, and cultured with HUVECs. Prostasomes were not shown to have a significant effect on HUVEC survival. Angiogenesis assays showed inhibition. The PKH-26-labelled particles were shown to have adhered to the HUVECs. This study adds the inhibition of an in vitro correlate of angiogenesis to the known actions of prostasomes.
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Neovascularización Patológica , Orgánulos/metabolismo , Próstata/citología , Enfermedades de la Próstata/fisiopatología , Técnicas de Cultivo de Célula , Células Endoteliales , Humanos , Masculino , Venas Umbilicales/citologíaRESUMEN
The association between cancer and thromboembolic disease is well known. This relationship is complex and in many, perhaps most, malignant diseases acts through multiple pathways. Increased tissue factor expression by endothelial cells, monocytes or macrophages has been implicated as one of these pathways. As well as being found in the circulating blood, tissue factor is also found in urine in a lipid-associated form. Although urinary tissue factor might be independent from that found in the circulation, its levels may reflect the status of peripheral blood monocyte activation. Thus, measurements of monocyte and/or urinary tissue factor may be of clinical significance, particularly in cancer, where levels could be valuable for diagnosis and monitoring progression.
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Neoplasias/metabolismo , Tromboplastina/metabolismo , Humanos , Curva ROCAsunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias/metabolismo , Tromboplastina/metabolismo , Adenoma/metabolismo , Adenoma/patología , Humanos , Neoplasias Intestinales/metabolismo , Neoplasias Intestinales/patología , Intestino Grueso/patología , Masculino , Neoplasias/patología , Pronóstico , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patologíaRESUMEN
Tissue factor (TF, coagulation factor III, CD142) is not only the main physiological initiator of normal blood coagulation, but is also important in the natural history of solid malignancies in that it potentiates metastasis and angiogenesis and mediates outside-in signalling. TF is expressed constitutively by many tissues which are not in contact with blood and by other cells upon injury or activation; the latter include endothelial cells, tissue macrophages, and peripheral blood monocytes. It can exist encrypted and unavailable functionally in the plasma membrane and the appearance of functional TF may be due to synthesis and/or de-encryption. Inflammatory cells often express TF and act to induce its production or de-encryption by other cells locally and, apparently, at remote sites. Inappropriate expression of TF by endothelial cells, macrophages or monocytes is thought to be an important trigger of coagulation in various pathological conditions. Several studies have shown that measurements of monocyte TF (mTF) may provide clinically significant information, particularly in patients with malignant and inflammatory diseases.
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Biomarcadores de Tumor/sangre , Monocitos/metabolismo , Neoplasias/sangre , Tromboplastina/metabolismo , Coagulación Sanguínea/fisiología , Regulación de la Expresión Génica/fisiología , Humanos , Tromboplastina/fisiologíaRESUMEN
Tissue factor (TF) is the main physiological initiator of blood coagulation and its activation or de-encryption within plasma membranes is important for trapping, extravasation and angiogenesis in the development and spread of solid malignancies. Multidrug resistance is also an adaptive response in malignant (and normal) cells. It is often mediated by the over-expression of the P-glycoprotein (P-gP) efflux pump. Both TF and P-gP tend to be expressed together, perhaps as part of a coherent 'crisis management' response of cells to environmental change or challenge. An associated feature in such a response appears to be the reversal of normal phospholipid charge asymmetry in the plasma membrane bilayer. Responses to environmental stimuli affect function in normal and malignant tissue. Uniting the study of TF expression or de-encryption and MDR-1 phenotype would be biologically enlightening and might ultimately influence clinical cancer management and the control of thrombotic problems associated with treatment.
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Resistencia a Múltiples Medicamentos , Resistencia a Antineoplásicos , Neoplasias/metabolismo , Tromboplastina/metabolismo , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Humanos , Neoplasias/patologíaRESUMEN
AIMS: Coagulation activation is a recognized complication of cancer in which increased tissue factor (TF) is implicated. TF can be detected in urine (uTF). This study assesses uTF levels in benign and malignant urological disease and correlates the results with conventional markers of tumour progression. METHODS: Using a simple and reproducible kinetic chromogenic assay, we determined uTF levels in controls (normal volunteers (n = 57) and patients with renal stones (n = 30)), benign and malignant bladder (n = 75) or prostate (n = 106) disease and in patients with or without recurrent bladder cancer (n=30). Each benign disease group was stratified as inflammatory (cystitis or prostatitis) or non-inflammatory (negative cystoscopy following haematuria or benign prostatic hypertrophy). RESULTS: The controls and the benign non-inflammatory results were indistinguishable. The malignant and inflammatory groups showed raised uTF levels over controls (P<0.001 bladder and P<0.01 prostate). The difference between malignant and benign inflammatory disease was only significant for the bladder group. uTF levels were significantly related to histological tumour grading, prostate serum specific antigen, static bone scan images and recurrence status. CONCLUSIONS: uTF levels can distinguish, statistically but not without overlap, patients with malignancy from normal controls and benign non-inflammatory conditions. Discrimination between inflammatory and malignant disease has only been demonstrated in the bladder. uTF levels showed a significant association with markers of tumour progression or metastasis and may be useful in predicting bladder tumour recurrence.
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Biomarcadores de Tumor/orina , Neoplasias de la Próstata/orina , Tromboplastina/orina , Neoplasias de la Vejiga Urinaria/orina , Adulto , Anciano , Neoplasias Óseas/secundario , Neoplasias Óseas/orina , Estudios de Casos y Controles , Cistitis/orina , Progresión de la Enfermedad , Humanos , Cálculos Renales/orina , Masculino , Persona de Mediana Edad , Antígeno Prostático Específico/sangre , Hiperplasia Prostática/orina , Neoplasias de la Próstata/inmunología , Neoplasias de la Próstata/patología , Prostatitis/orina , Neoplasias de la Vejiga Urinaria/inmunología , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
OBJECTIVE: The association between cancer and thromboembolic disease has been known for over a century. Increased tissue factor expression by endothelial cells, monocytes or macrophages is implicated. Thus, monocyte tissue factor measurements may reflect disease presence or progression. METHODS: Using a 2 stage kinetic chromogenic assay, monocyte tissue factor levels were assessed in normal controls (n=60), patient controls (hernia or cholecystectomy, n=60) and in patients with benign and malignant disease of the bladder (n=73), prostate (n=81), breast (n=83) and colorectum (n=62). This was performed as baseline (resting cells) and after 6 hours incubation with (stimulated) and without (unstimulated) lipopolysaccharide. Each benign disease group was sub-divided into inflammatory and non-inflammatory categories. RESULTS: The relative operating characteristic curve for the lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer, the area under the curve being 0.71. The control groups and the benign non-inflammatory groups gave similar results and were pooled for further analysis. Each malignant group showed higher monocyte tissue factor levels than the control groups for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). All benign inflammatory groups apart from breast, showed increased monocyte tissue factor levels over controls for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). In all cases there was no significant difference between the malignant and the benign inflammatory groups. Monocyte tissue factor levels were related to tumor grade or stage, patients' survival time, serum prostate specific antigen and static bone scan images. Levels were also higher in patients with bladder cancer recurrence and in those who subsequently died. CONCLUSION: Lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer compared to controls. Monocyte tissue factor levels are raised in malignant groups compared to controls and non-inflammatory diseases but not when compared with inflammatory conditions. Stimulated cells give better discrimination between the groups and may be useful in identifying high risk individuals. Monocyte tissue factor levels were related to tumor progression.
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Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/sangre , Neoplasias Colorrectales/sangre , Monocitos/metabolismo , Neoplasias de la Próstata/sangre , Tromboplastina/metabolismo , Neoplasias de la Vejiga Urinaria/sangre , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades de la Mama/sangre , Neoplasias de la Mama/complicaciones , Estudios de Casos y Controles , Enfermedades del Colon/sangre , Neoplasias Colorrectales/complicaciones , Análisis Discriminante , Progresión de la Enfermedad , Femenino , Humanos , Inflamación , Masculino , Persona de Mediana Edad , Enfermedades de la Próstata/sangre , Neoplasias de la Próstata/complicaciones , Factores de Riesgo , Sensibilidad y Especificidad , Tromboembolia/etiología , Enfermedades de la Vejiga Urinaria/sangre , Neoplasias de la Vejiga Urinaria/complicacionesRESUMEN
BACKGROUND: Abnormalities in laboratory coagulation and fibrinolysis parameters can be detected in cancer patients, and tissue factor (TF) is implicated. TF is produced by certain tumors and is increased in both tumor associated macrophages and blood monocytes (mTF). TF is also found in urine (uTF), and its levels may be clinically important. MATERIALS AND METHODS: Using a simple and highly standardized kinetic chromogenic assay (KCA), we measured uTF levels in controls (normal, n=57; patients with renal stones, n=30), patients with benign and malignant conditions of the bladder (n=75), prostate (n=106), breast (n=94), and colorectum (n=62). Each benign disease group was subdivided into inflammatory and noninflammatory categories. RESULTS: The controls and benign noninflammatory groups gave similar results and were, therefore, unified for further analysis. The malignant and inflammatory groups showed higher uTF levels than the controls (P<0.001 for bladder, P<0.01 prostate, P<0.001 breast, and P<0.001 for colorectum). The difference between malignant and benign inflammatory disease was significant for the bladder group (P<0.05). Cancer patients showed uTF activity above the upper quartile range of the normal control group--74.4% for bladder, 68.0% for prostate, 77.3% for breast and 73.0% for colorectal disease. uTF levels were related to tumor progression, patients survival time, serum prostate specific antigen (PSA), and static bone scan images (SBSI). Levels were also higher in patients with bladder cancer recurrence and those who subsequently died. CONCLUSION: uTF levels are raised in malignant and inflammatory disease compared to controls and patients with noninflammatory conditions, and are related to tumor grade or stage, patients survival and to markers of tumor progression.
RESUMEN
BACKGROUND: Activation of blood coagulation is a common complication of cancer and inflammation in both humans and experimental animals. Increased production of tissue factor--the principal initiator of the coagulation process--by endothelial cells, monocytes, and macrophages has been implicated in these conditions. AIM: To investigate whether urinary tissue factor (uTF) might reflect the state of monocyte/macrophage activation and be a useful diagnostic test. METHODS: Urine was centrifuged at 51,000 g to sediment tissue factor containing membrane vesicles. The tissue factor was then solubilised in beta-octyl-glucopyranoside and assayed in a specific chromogenic assay adapted for use in microtitre plates. RESULTS: The assay proved to be sensitive, specific, and reproducible. The normal range of uTF was relatively narrow and unaffected by age, sex, or cigarette smoking. Levels were not significantly influenced by storage of urine samples before assay or by the presence of fresh blood in the urine sample. CONCLUSIONS: This method may have diagnostic application in the study of haemostatic activation in patients with cancer and other disease states.
Asunto(s)
Biomarcadores de Tumor/orina , Neoplasias/diagnóstico , Tromboplastina/orina , Coagulación Sanguínea , Humanos , Activación de Linfocitos , Activación de Macrófagos , Neoplasias/inmunología , Neoplasias/orina , Valor Predictivo de las Pruebas , Reproducibilidad de los ResultadosRESUMEN
Monocytes express tissue factor (mTF) in several conditions including cancer where levels may be valuable in assessing tumour presence and progression. Using a two-stage kinetic chromogenic assay (KCA), mTF levels were measured in controls [normal subjects (n = 60) and patients undergoing hernia repair or cholecystectomy (n = 60)], in patients with benign and malignant disease of the breast (n = 83) and of the large bowel (n = 62). This was performed under fresh (resting) conditions and after incubation for 6 h without (unstimulated) and with (stimulated) Escherichia coli endotoxin. The malignant groups showed higher mTF levels than each of the three controls for resting (P < 0.05 breast, P < 0.05 colorectal) unstimulated (P < 0.05 breast, P < 0.05 colorectal) and stimulated cells (P < 0.001 breast, P < 0.01 colorectal). Similarly, the benign inflammatory groups had higher mTF levels than controls for resting (P < 0.05 colorectal), unstimulated (P < 0.05 colorectal) and stimulated cells (P < 0.01 breast, P < 0.01 colorectal). There was no significant difference between malignant and benign inflammatory groups in each organ. mTF levels showed an increase corresponding to that of histological tumour progression and were higher in non-surviving patients. In conclusion, mTF levels are raised in malignant and inflammatory disease compared to controls and patients with non-inflammatory conditions. Stimulated cells give better discrimination between the groups and may be of value in identifying high risk individuals. mTF levels showed an association with tumour grade or stage and the patients' survival time.
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Neoplasias de la Mama/metabolismo , Quimiocina CCL2/metabolismo , Neoplasias Colorrectales/metabolismo , Neoplasias de la Mama/patología , Neoplasias Colorrectales/patología , Progresión de la Enfermedad , Humanos , Estadificación de Neoplasias , Análisis de SupervivenciaRESUMEN
Activation of blood coagulation is a common complication of cancer in man and experimental animals. The causes of such activation may be multifactorial, but increased production of tissue factor (TF) by the host mononuclear cells may be involved. TF is not only produced by human monocytes (mTF) and tumour cells, but is also found in urine (uTF), where measurements might be clinically important. Using a highly reproducible (intra-assay CV 2.3 per cent and inter-assay CV 8.1 per cent) one-stage kinetic chromogenic assay (KCA) developed by this group, uTF levels were measured in controls [healthy volunteers (n = 57), patients with renal stones and a normal ESR (n = 30)] and in patients with benign and malignant diseases of the breast (n = 94) and large bowel (n = 62). Each benign disease group was sub-divided into inflammatory and non-inflammatory categories. There were no significant differences between the controls and the benign non-inflammatory groups, so they were unified for further analysis. Malignant groups, irrespective of tumour types, showed significantly higher uTF levels than controls (p < 0.001 for breast and p < 0.01 for large bowel). Similarly, breast and colorectal benign inflammatory groups showed significant increases over controls (p < 0.01 and p < 0.001, respectively). Patients with malignant disease showed uTF activity above the upper quartile range of the normal control group for breast, 77.3 per cent, and large bowel, 73 per cent. uTF levels were related to histological tumour grading and were higher in non-surviving patients. In conclusion, uTF levels are raised in malignant and inflammatory disease compared with controls and patients with non-inflammatory conditions. uTF levels may reflect tumour progression.
Asunto(s)
Biomarcadores de Tumor/orina , Neoplasias de la Mama/orina , Neoplasias Colorrectales/orina , Proteínas de Neoplasias/orina , Tromboplastina/orina , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades de la Mama/orina , Neoplasias de la Mama/patología , Niño , Preescolar , Neoplasias Colorrectales/patología , Diagnóstico Diferencial , Femenino , Humanos , Cálculos Renales/orina , Masculino , Persona de Mediana Edad , Tasa de SupervivenciaRESUMEN
Tissue factor (TF) is the main physiological initiator of blood coagulation and may be important in the biology of a variety of solid malignancies, particularly where angiogenesis is a critical factor. TF is frequently encrypted in the plasma membrane of cells in contact with blood, and is exposed only after stimulation by certain agonists. Cancer cells variably express TF and cancer cell lines which exhibit multidrug resistance contain more TF than parental cells. TF is increased in both tumour-associated macrophages and blood monocytes and has been implicated in abnormal coagulation activation seen in patients with inflammatory conditions and cancer. TF is also found in urine (uTF) in a lipid-associated form, probably of kidney origin. uTF levels can be assayed in a cost-effective manner and may be clinically important, particularly in patients with renal disorders and malignancy. uTF levels are not significantly affected by age, gender or cigarette smoking.
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Biomarcadores de Tumor/orina , Enfermedades Renales/orina , Tromboplastina/orina , Factores de Edad , Coagulación Sanguínea , Resistencia a Antineoplásicos , Ensayo de Inmunoadsorción Enzimática , Humanos , Riñón/metabolismo , Enfermedades Renales/diagnóstico , Neoplasias Renales/diagnóstico , Neoplasias Renales/orina , Factores Sexuales , Tromboplastina/metabolismoRESUMEN
BACKGROUND: We have previously explored the clinical significance of urinary tissue factor (uTF) in patients with glomerulonephritis (GN) and malignancy. However, the functional and structural properties and putative cell of origin of uTF are poorly documented. In these studies we investigate these aspects of uTF. METHODS: Functional and structural properties of uTF were investigated using a one stage kinetic chromogenic assay, an enzyme-linked immunoabsorbent assay (ELISA) and transmission electron microscopy (TEM) on urine samples collected from healthy controls (n=69). The distribution of uTF and anionic phospholipid in the kidney were studied in sections from normal areas of nephrectomy specimens, using an immunoperoxidase technique. These were stained for tissue factor (TF) antigen and recombinant Annexin V. RESULTS: We found uTF to be present on subcellular particles as visualized by TEM. These particles contained anionic phospholipid as evidenced by binding to Annexin V fluorescein isothiocyanate (FITC). Although TF is present in urine in a functional and antigenic form no association was observed between the two. Using immunoperoxidase-based techniques, the cytoplasm of both distal and proximal tubules (but not glomerular cells) was positive for TF antigen and Annexin V. CONCLUSION: uTF is found on subcellular particles which provide lipid for its functional activity. Both uTF and its associated vesicles are found in the renal tubular cells.