RESUMO
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.
Assuntos
Metaloproteinase 9 da Matriz/sangue , Tuberculose Pulmonar/enzimologia , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Metaloproteinase 2 da Matriz/sangue , Pessoa de Meia-Idade , Índice de Gravidade de DoençaRESUMO
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.
Assuntos
Biomarcadores Tumorais/análise , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma de Células Pequenas/genética , Carcinoma de Células Pequenas/patologia , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Idoso , Feminino , Humanos , Imuno-Histoquímica , Masculino , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Pessoa de Meia-Idade , Neovascularização Patológica , FenótipoRESUMO
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.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma de Células Pequenas/enzimologia , Neoplasias Pulmonares/enzimologia , Metaloproteinase 9 da Matriz/sangue , Adulto , Idoso , Feminino , Humanos , Masculino , Metaloproteinase 2 da Matriz/sangue , Pessoa de Meia-IdadeRESUMO
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.
Assuntos
Precursores de Proteínas/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Taquicininas/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Clonagem Molecular , Escherichia coli/genética , Expressão Gênica , Genes Sintéticos/genética , Dados de Sequência Molecular , Precursores de Proteínas/química , Precursores de Proteínas/genética , Sinais Direcionadores de Proteínas/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Taquicininas/química , Taquicininas/genéticaAssuntos
Encéfalo/metabolismo , Metaloendopeptidases/metabolismo , Substância P/metabolismo , Animais , Encéfalo/enzimologia , Dipeptidil Peptidases e Tripeptidil Peptidases/metabolismo , Humanos , Técnicas In Vitro , Neprilisina/metabolismo , Fosforilação Oxidativa , Peptidil Dipeptidase A/metabolismo , Ratos , Sinapsinas/metabolismoAssuntos
Encéfalo/metabolismo , Neurocinina A/biossíntese , Neurocinina B/biossíntese , Neuropeptídeos/biossíntese , Medula Espinal/metabolismo , Substância P/biossíntese , Animais , Bovinos , Humanos , Neurocinina A/genética , Neurocinina B/genética , Neuropeptídeos/genética , Biossíntese de Proteínas , Ratos , Substância P/genética , Suínos , Transcrição GênicaAssuntos
Quimiotaxia de Leucócito/imunologia , Inflamação/imunologia , Neutrófilos/imunologia , Receptores de Neurotransmissores/fisiologia , Substância P/fisiologia , Sequência de Aminoácidos/genética , Aminoácidos/química , Aminoácidos/genética , Humanos , Dados de Sequência Molecular , Receptores da Neurocinina-1 , Substância P/química , Substância P/genéticaRESUMO
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.
Assuntos
Cálcio/fisiologia , Calmodulina/fisiologia , Córtex Cerebral/metabolismo , Proteínas de Membrana/metabolismo , Substância P/farmacologia , Sinaptossomos/metabolismo , Animais , Autorradiografia , Córtex Cerebral/efeitos dos fármacos , Eletroforese em Gel de Poliacrilamida , Técnicas In Vitro , Radioisótopos de Fósforo , Fosforilação , Ratos , Ratos Endogâmicos , Sinapsinas/metabolismo , Sinaptossomos/efeitos dos fármacosAssuntos
Química Encefálica , Encéfalo/metabolismo , Neurônios/metabolismo , Proteínas Quinases/metabolismo , Sinapsinas/metabolismo , Encéfalo/citologia , Encéfalo/enzimologia , Catálise , Humanos , Neurônios/enzimologia , Fosforilação Oxidativa , Biossíntese de Proteínas/genética , Biossíntese de Proteínas/fisiologia , Proteínas Quinases/genética , Sinapsinas/análise , Sinapsinas/genética , Transcrição Gênica/genética , Transcrição Gênica/fisiologiaRESUMO
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.
