RESUMEN
SETTING: Parenchymal lung destruction accompanied by active tuberculosis is, at least in part, caused by host as well as bacillus metalloproteinases. Mycobacterium tuberculosis has been shown to stimulate MMP-9 expression in the lung of infected organisms. DESIGN: We have used quantitative zymography and computer-assisted image analysis to measure the levels of type IV collagenases in 20 serum samples of patients with active tuberculosis and in 23 serum samples of healthy volunteers. RESULTS: Mean levels of the serum MMP-9 were over three-fold higher in tuberculous samples compared with normal serum (P < 0.0001), whereas the MMP-2 levels did not differ in these two groups. The levels of MMP-9 were significantly higher in subjects with advanced disease than in those with only limited disease changes (P < 0.05). CONCLUSIONS: We suppose that the elevation of serum MMP-9 levels in patients with tuberculosis is affected by the augmentation of synthesis and/or secretion of this enzyme by inflammatory cells in response to M. tuberculosis infection. The observed association between the serum MMP-9 level and the extent of radiological change suggests that the quantification of the serum level of this enzyme may constitute a supplementary test in pulmonary tuberculosis diagnostics.
Asunto(s)
Metaloproteinasa 9 de la Matriz/sangre , Tuberculosis Pulmonar/enzimología , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Metaloproteinasa 2 de la Matriz/sangre , Persona de Mediana Edad , Índice de Severidad de la EnfermedadRESUMEN
PURPOSE: Matrix metalloproteinases MMP-2 and MMP-9 are implicated in invasion and metastasis of malignant tumors. We investigated the expression and activation of MMP-2 and MMP-9 in lung cancer compared with normal lung parenchyma, and looked for a potential marker of malignancy. METHODS: Thirty-six pulmonary carcinomas and paired normal lung specimens were analyzed by gelatin zymography and computer-assisted image analysis for the expression of MMP-2 and MMP-9. RESULTS: We showed that expression of both type IV collagenases was remarkably higher in carcinoma samples than in lung parenchyma. The MMP-9 levels in lung cancer were over twofold higher than in normal lung tissues. The levels of latent and active forms of MMP-2 in lung cancer samples were, correspondingly, 3.8- and 17-fold higher than in lung parenchyma. The tumor/normal (T/N) ratios of MMP-2 were negatively correlated with the hemoglobin levels and erythrocytes number. CONCLUSIONS: A high level of the active form of MMP-2 in almost all of the carcinomas and the near lack of its activation in normal lung parenchyma shows that MMP-2 activation is associated with the malignant phenotype and may serve as a good marker of malignancy. The correlation between low hemoglobin level and T/N ratio of MMP-2 may indicate significance of MMP-2 for angiogenesis.
Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Células Pequeñas/genética , Carcinoma de Células Pequeñas/patología , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Metaloproteinasa 2 de la Matriz/biosíntesis , Metaloproteinasa 9 de la Matriz/biosíntesis , Anciano , Femenino , Humanos , Inmunohistoquímica , Masculino , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/análisis , Persona de Mediana Edad , Neovascularización Patológica , FenotipoRESUMEN
The 72 kDa matrix metalloproteinase (MMP-2) and the 92 kDa matrix metalloproteinase (MMP-9), are type IV collagenases that have been implicated as important factors in cancer invasion and metastasis formation. We have used quantitative zymography and computer-assisted image analysis to measure the levels of MMP-9 and MMP-2 in 19 samples of serum of lung cancer patients and in 23 samples of normal serum. Mean levels of MMP-9 were significantly elevated in cancer samples compared with normal sera (1.33 +/- 0.61 microU microl(-1) vs. 0.37 +/- 0.10 microU microl(-1), P<0.0001). MMP-2 levels did not differ significantly in these two groups. However, there was no significant correlation between serum MMP-9 activity and the disease stage. We found that circulation levels of MMP-9 in lung cancer patients is 3.6-fold higher than in healthy volunteers, however, we do not consider this elevation to be a direct reflection of MMP-9 over-production by tumour cells.
Asunto(s)
Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/enzimología , Carcinoma de Células Pequeñas/enzimología , Neoplasias Pulmonares/enzimología , Metaloproteinasa 9 de la Matriz/sangre , Adulto , Anciano , Femenino , Humanos , Masculino , Metaloproteinasa 2 de la Matriz/sangre , Persona de Mediana EdadRESUMEN
Using an efficient Escherichia coli expression system, we have been able to obtain the precursor of substance P, alpha-preprotachykinin (alpha PPT). The alpha PPT protein is produced in E. coli as a fusion to beta-galactosidase, and accumulates in the cytoplasm as insoluble inclusion bodies. We also produced protachykinin (alpha PT), i.e., alpha PPT without a signal peptide. Further purification and characterization of the alpha PPT and alpha PT polypeptides strongly suggest that fully purified products can be obtained using our procedures.
Asunto(s)
Precursores de Proteínas/biosíntesis , Proteínas Recombinantes de Fusión/biosíntesis , Taquicininas/biosíntesis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Clonación Molecular , Escherichia coli/genética , Expresión Génica , Genes Sintéticos/genética , Datos de Secuencia Molecular , Precursores de Proteínas/química , Precursores de Proteínas/genética , Señales de Clasificación de Proteína/genética , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Taquicininas/química , Taquicininas/genéticaAsunto(s)
Encéfalo/metabolismo , Metaloendopeptidasas/metabolismo , Sustancia P/metabolismo , Animales , Encéfalo/enzimología , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/metabolismo , Humanos , Técnicas In Vitro , Neprilisina/metabolismo , Fosforilación Oxidativa , Peptidil-Dipeptidasa A/metabolismo , Ratas , Sinapsinas/metabolismoAsunto(s)
Encéfalo/metabolismo , Neuroquinina A/biosíntesis , Neuroquinina B/biosíntesis , Neuropéptidos/biosíntesis , Médula Espinal/metabolismo , Sustancia P/biosíntesis , Animales , Bovinos , Humanos , Neuroquinina A/genética , Neuroquinina B/genética , Neuropéptidos/genética , Biosíntesis de Proteínas , Ratas , Sustancia P/genética , Porcinos , Transcripción GenéticaAsunto(s)
Quimiotaxis de Leucocito/inmunología , Inflamación/inmunología , Neutrófilos/inmunología , Receptores de Neurotransmisores/fisiología , Sustancia P/fisiología , Secuencia de Aminoácidos/genética , Aminoácidos/química , Aminoácidos/genética , Humanos , Datos de Secuencia Molecular , Receptores de Neuroquinina-1 , Sustancia P/química , Sustancia P/genéticaAsunto(s)
Química Encefálica , Encéfalo/metabolismo , Neuronas/metabolismo , Proteínas Quinasas/metabolismo , Sinapsinas/metabolismo , Encéfalo/citología , Encéfalo/enzimología , Catálisis , Humanos , Neuronas/enzimología , Fosforilación Oxidativa , Biosíntesis de Proteínas/genética , Biosíntesis de Proteínas/fisiología , Proteínas Quinasas/genética , Sinapsinas/análisis , Sinapsinas/genética , Transcripción Genética/genética , Transcripción Genética/fisiologíaRESUMEN
1. We have used synaptosomal membranes to study the influence of substance P and its fragments and analogues of its C-terminal fragment on Ca2+/calmodulin-dependent synapsin I endogenous phosphorylation. 2. SP1-11, SP1-4, [Tyr8]SP6-11 and [pGlu6, Tyr8]SP6-11 at 10(-3) M greatly inhibited synapsin I phosphorylation. 3. SP6-11 at all investigated concentrations and SP1-11, SP1-4, [Tyr8]SP6-11, [pGlu6, Tyr8]SP6-11 at 10(-4) and 10(-5) M were ineffective. 4. The results indicate that SP1-11 and its N-terminal fragment and analogues of its C-terminal fragment act on the phosphorylation of specific synaptic protein (synapsin I) and therefore may influence the release of neurotransmitters, membrane conductance and potentiation or inhibition of other signalling systems.
Asunto(s)
Calcio/fisiología , Calmodulina/fisiología , Corteza Cerebral/metabolismo , Proteínas de la Membrana/metabolismo , Sustancia P/farmacología , Sinaptosomas/metabolismo , Animales , Autorradiografía , Corteza Cerebral/efectos de los fármacos , Electroforesis en Gel de Poliacrilamida , Técnicas In Vitro , Radioisótopos de Fósforo , Fosforilación , Ratas , Ratas Endogámicas , Sinapsinas/metabolismo , Sinaptosomas/efectos de los fármacosRESUMEN
1. The effects of substance P and its fragments and analogue of a C-terminal fragment on cyclic AMP-dependent phosphorylation of synapsin I in synaptosomal membranes (SM) from cerebral cortex were investigated. 2. SP(I-II) and SP(1-4) at 10(-3) M caused a marked stimulation of synapsin I phosphorylation. 3. A C-terminal fragment of SP (SP6-11) had no effect on phosphorylation of synapsin 1. 4. Analogue of C-terminal fragment [(Tyr8)SP6-11] at 10(-3) M distinctly inhibits phosphorylation of synapsin I. 5. These data suggest that SPI-II and its C- and N-terminal fragments have a modulator function against the phosphorylation of some rat brain proteins.