RESUMO
BACKGROUND: The development of specific screening programs for individuals with a family history of colorectal cancer (CRC) is a priority. This study evaluates the diagnostic performance of serum soluble CD26 (sCD26) in family-risk individuals and compares this marker with the faecal immunochemical test for the detection of advanced neoplasia (AN) (CRC or advanced adenomas; AA). METHODS: Five hundred and sixteen asymptomatic individuals with at least one first-degree relative with CRC were included. Serum sCD26 was measured in all the individuals who also underwent a colonoscopy (53 AA and four cancer cases were found) and a faecal immunochemical test. RESULTS: Setting specificity to 90% and 95%, respectively, sCD26 showed a sensitivity of 39.6% and 28.3% for AA, and of 42.1% and 28.1% for AN. The combination of sCD26 and the faecal test detected AA and AN with a 52.8% and 56.1% sensitivity, corresponding to 93.5% specificity. CONCLUSIONS: The combination of serum sCD26 and the faecal blood test could result a valuable strategy for detecting AN in familial-risk CRC screening.
Assuntos
Adenoma/diagnóstico , Biomarcadores Tumorais/sangue , Neoplasias Colorretais/diagnóstico , Dipeptidil Peptidase 4/sangue , Sangue Oculto , Adenoma/sangue , Adenoma/enzimologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/sangue , Neoplasias Colorretais/enzimologia , Método Duplo-Cego , Detecção Precoce de Câncer , Feminino , Humanos , Imunoensaio , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Risco , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Novel non-invasive biomarkers for the precise diagnosis of malignancy in pleural effusion (PE) are needed. The aim of this study was to determine the diagnostic accuracy of calprotectin for predicting malignancy in patients with exudative PE. METHODS: Calprotectin concentration was measured in 156 individuals diagnosed with exudative PE (67 malignant and 89 benign). Calprotectin accuracy for discriminating between malignant and benign PE was evaluated using receiver operating characteristic (ROC) curves. Univariate and multivariate logistic regression were performed to test the association between calprotectin levels and malignant PE. RESULTS: Calprotectin levels were significantly lower in malignant pleural fluid (257.2 ng ml(-1), range: 90.7-736.4) than benign effusions (2627.1 ng ml(-1), range: 21-9530.1). The area under the curve was 0.963. A cutoff point of ≤ 736.4 ng ml(-1) rendered a sensitivity of 100%, with a specificity of 83.15%, which could prove useful to delimit those patients with negative cytology tests that should be referred for more invasive diagnostic procedures. Logistic regression demonstrated a strong association between calprotectin and malignancy (adjusted OR 663.14). CONCLUSION: Calprotectin predicts malignancy in pleural fluid with high accuracy and could be a good complement to cytological methods.
Assuntos
Complexo Antígeno L1 Leucocitário/análise , Derrame Pleural Maligno/diagnóstico , Derrame Pleural/diagnóstico , Adulto , Idoso , Biomarcadores/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Curva ROCRESUMO
Butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) display both esterase and aryl acylamidase (AAA) activities. Their AAA activity can be measured using o-nitroacetanilide (ONA). In human samples depleted of acetylcholinesterase, we noticed that the ratio of amidase to esterase activities varied depending on the source, despite both activities being due to BuChE. Searching for an explanation, we compared the activities of BuChE molecular forms in samples of human colon, kidney and serum, and observed that BuChE monomers (G(1)) hydrolyzed o-nitroacetanilide much faster than tetramers (G(4)). This fact suggested that association might cause differences in the AAA site between single and polymerized subunits. This and other post-translational modifications in BuChE subunits probably determine their level of AAA activity. The higher amidase activity of monomers could justify the presence of single BuChE subunits in cells as a way to preserve the AAA activity of BuChE, which could be lost by oligomerization.
Assuntos
Aminopeptidases/metabolismo , Butirilcolinesterase/metabolismo , Colo/enzimologia , Neoplasias Colorretais/enzimologia , Humanos , Reto/enzimologiaRESUMO
The discovery of kinase domain mutations in the epidermal growth factor receptor gene (EGFR) in never-smoker patients, associated with an increased sensitivity to tyrosine kinase inhibitors (TKIs) such as gefitinib or erlotinib, has been one of the most relevant findings ever in non-small cell lung carcinomas (NSCLC). Since treatment with TKIs has furthermore shown a clinical benefit in head and neck squamous cell carcinoma (HNSCC) patients, we hypothesized that these mutations could also be present in this neoplasia. Current studies looking for EGFR mutations in HNSCC are limited and results are still controversial. In this work, we screened for EGFR tyrosine kinase mutations in tumour DNA obtained from 31 Spanish patients with HNSCC by PCR-single-strand conformational polymorphism analysis. None of the patients displayed a somatic EGFR mutation, previously described in NSCLC, but other DNA sequence variations were found in 9 of 31 HNSCC patients. Accordingly, activating EGFR mutations in HNSCC patients seem to be a rare event in Spanish patients, suggesting that there is little room for the administration of TKIs in HNSCC based on the presence of these mutations. Additional investigations about EGFR amplification are indicated to establish a potential relationship between EGFR overexpression and the response to anti-EGFR therapies.
Assuntos
Carcinoma de Células Escamosas/genética , Receptores ErbB/química , Genes erbB-1/genética , Neoplasias de Cabeça e Pescoço/genética , Proteínas de Neoplasias/química , Idoso , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/epidemiologia , Análise Mutacional de DNA , DNA de Neoplasias/genética , Receptores ErbB/antagonistas & inibidores , Neoplasias de Cabeça e Pescoço/enzimologia , Neoplasias de Cabeça e Pescoço/epidemiologia , Humanos , Neoplasias Laríngeas/enzimologia , Neoplasias Laríngeas/epidemiologia , Neoplasias Laríngeas/genética , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/enzimologia , Neoplasias Bucais/epidemiologia , Neoplasias Bucais/genética , Proteínas de Neoplasias/antagonistas & inibidores , Neoplasias Faríngeas/enzimologia , Neoplasias Faríngeas/epidemiologia , Neoplasias Faríngeas/genética , Polimorfismo Conformacional de Fita Simples , Inibidores de Proteínas Quinases/uso terapêutico , Fumar/epidemiologia , Fumar/genética , Espanha/epidemiologiaRESUMO
Serum levels of the soluble epidermal growth factor receptor (sEGFR) and its ligands epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha) and amphiregulin (AR) were measured in healthy donors and patients with non-small cell lung cancer (NSCLC) and head and neck carcinoma (HNC). In NSCLC, we found sEGFR and EGF levels significantly lowered in patients with respect to healthy donors. In HNC patients, significantly diminished levels were found in the case of sEGFR, EGF and also AR. In both malignancies, no significant association was found between the serum levels of the molecules and the patients' gender, age or smoking habit. Only a significant association was found between the decrease of sEGFR and the absence of distant metastasis in NSCLC and the tumour stage in HNC. The most interesting result was that combining sEGFR and EGF, sensitivities of 88% in NSCLC and 100% in HNC were reached without losing specificity (97.8% in both cases). The use of discriminant analysis and logistic regression improved the sensitivity for NSCLC and the specificity for HNC. These data demonstrate a potentially interesting value of the serum levels of sEGFR and EGF, especially when combined, as markers for NSCLC and HNC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma/sangue , Fator de Crescimento Epidérmico/sangue , Receptores ErbB/sangue , Neoplasias de Cabeça e Pescoço/sangue , Neoplasias Pulmonares/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/sangue , Feminino , Humanos , Ligantes , Masculino , Pessoa de Meia-Idade , Receptores Androgênicos/sangue , Fator de Crescimento Transformador alfa/sangueRESUMO
This study was conducted to evaluate the significance of preoperative serum sialic acid levels in the diagnosis and prognosis of colorectal cancer (CRC). Total sialic acid (TSA) was determined by the thiobarbituric acid method and normalized to total protein (TP). A postoperative follow-up of CRC patients classified as Dukes' stages A, B or C was performed and survival analysis was carried out to evaluate the impact of sialic acid levels on tumor recurrence. Our diagnostic studies indicate that TSA/TP is a better marker than either TSA or carcinoembryonic antigen (CEA), especially for the detection of CRC patients at an early stage. At a cutoff of 30.90 nmol/mg of protein, TSA/TP showed a sensitivity of 85% with a specificity of 97% to discriminate CRC patients from healthy donors. In survival analysis, both TSA and TSA/TP were found to be significant prognostic factors for tumor recurrence in CRC. Furthermore, TSA/TP could distinguish patients at high risk of recurrence within Dukes' stage B and in multivariate analysis it was identified as the best independent prognostic factor. According to our results, preoperative serum TSA/TP content could supply additional information to that provided by Dukes' stage about the prognosis of CRC patients.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Colorretais/sangue , Ácido N-Acetilneuramínico/sangue , Neoplasias/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno Carcinoembrionário/sangue , Estudos de Casos e Controles , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/patologia , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Modelos de Riscos Proporcionais , Recidiva , Sensibilidade e Especificidade , Substâncias Reativas com Ácido Tiobarbitúrico , Fatores de TempoRESUMO
The acid alpha-L-fucosidase is usually found as a soluble component of lysosomes where fucoglycoconjugates are degraded. In the present investigation, we have demonstrated the existence of a cell surface protein with enzymatic alpha-L-fucosidase activity that crossreacts specifically with a rabbit anti-(alpha-L-fucosidase) Ig. By different approaches, this alpha-L-fucosidase, which represents 10-20% of the total cellular fucosidase activity, was detected in all the tested human cells (hemopoietic, epithelial, mesenchymal). Two bands of approximately 43-49 kDa were observed, although theoretical data support the possibility of having the same genetic origin that the known 50 to 55-kDa Mr alpha-L-fucosidase. We speculate about an alternative traffic pathway for the plasma membrane alpha-L-fucosidase to work on the rapid turnover of glycoproteins.
Assuntos
Membrana Celular/enzimologia , alfa-L-Fucosidase/análise , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Western Blotting , Fracionamento Celular , Linhagem Celular , Reagentes de Ligações Cruzadas , Eletroforese em Gel de Poliacrilamida , Células Epiteliais/enzimologia , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Linfócitos/enzimologia , Monócitos/enzimologia , Neutrófilos/enzimologia , Testes de Precipitina , alfa-L-Fucosidase/imunologia , alfa-L-Fucosidase/metabolismoRESUMO
OBJECTIVES: The purpose of this study was to assess the value of the serum levels of alpha-L-fucosidase activity in the diagnosis of patients with colorectal cancer. METHODS: Using a fluorometric method we analyzed the alpha-L-fucosidase activity in preoperative sera from 137 colorectal cancer patients and in sera from 232 donors. RESULTS: The enzymatic activity of alpha-L-fucosidase was significantly lower (p <0.001) in patients (4.8+/-3.09 U/ml) than in donors (10.5+/-5.46 U/ml). Using the ROC curve, the ideal cut-off for the diagnostic value of alpha-L-fucosidase activity was determined to be 5.6 U/ml. The diagnostic efficiency for colorectal cancer of alpha-L-fucosidase activity was higher than that observed for carcinoembryonic antigen (cut-off 5.0 ng/ml), especially for tumors at an early stage. CONCLUSIONS: Our results suggest that preoperative serum alpha-L-fucosidase activity may be used as a cheap and easy complementary test, in addition to standard clinical procedures routinely used for the diagnosis of colorectal cancer.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Colorretais/diagnóstico , alfa-L-Fucosidase/sangue , Antígeno Carcinoembrionário/sangue , Estudos de Casos e Controles , Neoplasias Colorretais/enzimologia , Diagnóstico Diferencial , Feminino , Humanos , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Sensibilidade e EspecificidadeRESUMO
The expression of different sialoglycoconjugates and fucoglycoconjugates in normal mucosa and adenocarcinoma samples from 43 colorectal cancer patients was investigated by using specific lectins and applying a semiquantitative analysis. A pronounced decrease in the intracellular binding of the Maackia amurensis lectin, which recognizes alpha(2,3)-linked sialic acid residues, was found in the tumoral tissue. In contrast, a significant increase in the staining with the Sambucus nigra lectin (SNA I), which binds to alpha(2,6)-linked sialic acid residues, was detected in the epithelial cells as well as in the mucins from tumors. No significant differences in the reactivity with the Aleuria aurantia lectin, which recognizes the sequence Fuc(alpha1,6)GlcNAc, between normal and malignant colorectal tissues were detected. Furthermore, the correlation between lectin-binding profiles and the prognosis of colorectal cancer patients was examined. After an average postoperative follow-up period of 31 months, patients with tumors showing a strong SNA I staining presented a greater probability of disease recurrence. This result suggests that the intensity of staining with SNA I could be a valid parameter for predicting recurrence in colorectal cancer.
Assuntos
Adenocarcinoma/química , Adenocarcinoma/diagnóstico , Neoplasias Colorretais/química , Neoplasias Colorretais/diagnóstico , Fucose/análise , Ácido N-Acetilneuramínico/análise , Lectinas de Plantas , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Antígenos Glicosídicos Associados a Tumores/análise , Antígenos Glicosídicos Associados a Tumores/metabolismo , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Seguimentos , Fucose/análogos & derivados , Fucose/metabolismo , Humanos , Imuno-Histoquímica , Mucosa Intestinal/química , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Lectinas/metabolismo , Ácido N-Acetilneuramínico/análogos & derivados , Ácido N-Acetilneuramínico/metabolismo , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Fito-Hemaglutininas/metabolismo , Prognóstico , Proteínas Inativadoras de Ribossomos , Estatísticas não Paramétricas , Especificidade por SubstratoRESUMO
The enhancement of lysosomal beta-hexosaminidase degradative activity in different human cancer tissues is fairly well documented. Gastric tumors have attracted considerable attention on the basis of their social incidence and clinical recurrence. Here we report a comparative study of beta-hexosaminidase activity and of its isoenzymes beta-hexosaminidase A (HA) and beta-hexosaminidase B (HB) from gastric adenocarcinoma and normal mucosa. Tumor beta-hexosaminidase activity from crude extracts and chromatographically resolved HA and HB forms were analyzed as regards their physicochemical and enzymatic properties and were compared to similar samples obtained from control tissue. The existence of one active site in the beta-hexosaminidase enzyme responsible for both N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D- galactosaminidase activities was determined. Apart from their relative contributions to beta-hexosaminidase activities, two major differences appeared in tumor HA and HB forms with respect to the corresponding controls: (1) the presence of an atypical heat-stable HB isoenzyme in gastric adenocarcinoma, and (2) a significantly increased Vmax of the HA form acting on both p-nitrophenyl-N-acetyl-beta-D-glucosaminide and p-nitrophenyl-N-acetyl-beta-D-galactosaminide substrates. The results show that the beta-hexosaminidase HA and HB isoenzymes from gastric adenocarcinoma display different patterns of response from the same forms from other human tumors.
Assuntos
Adenocarcinoma/enzimologia , Neoplasias Gástricas/enzimologia , beta-N-Acetil-Hexosaminidases/metabolismo , Acetilglucosaminidase/metabolismo , Cromatografia , Ativação Enzimática , Mucosa Gástrica/enzimologia , Temperatura Alta , Humanos , Concentração de Íons de Hidrogênio , Isoenzimas/metabolismo , CinéticaRESUMO
OBJECTIVES: We have carried out a detailed study of some glycosidases in an attempt to explain the differential profile of enzyme activity between human colonic adenocarcinoma and normal mucosa. DESIGN AND METHODS: Several glycosidase activities associated with human colonic adenocarcinoma and control tissues were submitted to a detailed structural and functional characterization. RESULTS: Tumoral and control samples were assayed for beta-D-galactosidase, beta-D-glucuronidase, alpha-D-mannosidase, beta-NAc-D-glucosaminidase and beta-NAc-D-galactosaminidase activities. Tumoral tissue showed higher beta-D-galactosidase, beta-NAc-D-glucosaminidase, and beta-NAc-D-galactosaminidase activities than control tissue. Glycosidases from tumoral and control tissues demonstrated no differences in optimum pH, subcellular distribution, pH and thermal stability. However, the kinetic analysis showed a statistically significant increased Vmax in tumoral colon with respect to the control for beta-D-galactosidase, beta-NAc-D-glucosaminidase, and beta-NAc-D-galactosaminidase activities. The Km remained unaltered. CONCLUSIONS: The increased Vmax detected for some glycosidase activities in human colonic adenocarcinoma could correspond with a greater presence of enzyme proteins in the tumoral cells, and not to changes in protein and/or active site structure.
Assuntos
Adenocarcinoma/enzimologia , Neoplasias do Colo/enzimologia , Glicosídeo Hidrolases/metabolismo , Acetilglucosaminidase/metabolismo , Adenocarcinoma/química , Adenocarcinoma/metabolismo , Neoplasias do Colo/química , Neoplasias do Colo/metabolismo , Ativação Enzimática , Estabilidade Enzimática , Glicosídeo Hidrolases/química , Temperatura Alta , Humanos , Concentração de Íons de Hidrogênio , Cinética , Manosidases/metabolismo , Frações Subcelulares/enzimologia , alfa-Manosidase , beta-Galactosidase/metabolismoRESUMO
Sialic acid levels were determined in serum and in both normal and tumour-derived tissues from 30 patients with colorectal cancer. Total sialic acid (TSA), bound sialic acid (BSA),TSA normalised to total protein (TSA/TP) and BSA normalised to total protein (BSA/TP) were significantly higher (P < 0.0001) in sera from patients than in normal subjects. We found a trend of increasing serum sialic acid levels (TSA/TP and BSA/TP) as the malignancy became more severe (i.e. Dukes' stages A to C). Comparison of sialic acid levels between normal and tumour-derived colorectal tissues indicated no statistically significant differences in TSA, BSA or FSA (free sialic acid) levels between both tissues; however, TSA/TP and BSA/TP values were significantly decreased (P < 0.05) in the tumoral tissue. In this study, the possible relation between serum and tumour sialic acid levels in colorectal cancer patients was investigated. Our results showed that in these patients there was no correlation between serum BSA and tumour BSA levels.
Assuntos
Adenocarcinoma/sangue , Adenocarcinoma/metabolismo , Neoplasias Colorretais/sangue , Neoplasias Colorretais/metabolismo , Ácidos Siálicos/sangue , Ácidos Siálicos/metabolismo , Adenocarcinoma/patologia , Proteínas Sanguíneas/metabolismo , Neoplasias Colorretais/patologia , Estudos de Avaliação como Assunto , Humanos , Proteínas de Neoplasias/sangue , Proteínas de Neoplasias/metabolismo , Estadiamento de Neoplasias , Valores de ReferênciaRESUMO
Fucose levels were determined in normal and tumour-derived tissues from patients with colorectal adenocarcinoma. Total, free and bound fucose were significantly higher (P < 0.001) in the tumoral tissue. Comparison of serum fucose levels between patients with colorectal cancer and control subjects indicated no statistically significant differences in total, free and bound fucose expressed as nmol/ml serum. However, when total and bound fucose were normalized to total protein (nmol/mg protein), an elevation in both parameters was found in colorectal cancer patients, although only that for total fucose was statistically significant (P < 0.001). A clear association between fucose content and clinical stage of patients according to the Dukes' classification can not be established, although patients at stage C showed higher levels of fucose both in serum and tissue. The current investigation provides direct evidence of a relationship between the elevation of fucose and the presence of tumour in colorectal cancer patients.
Assuntos
Adenocarcinoma/química , Neoplasias Colorretais/química , Fucose/análise , Adenocarcinoma/sangue , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/sangue , Neoplasias Colorretais/patologia , Feminino , Fucose/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Metástase Neoplásica , Estadiamento de NeoplasiasRESUMO
Human colon sialidase has been characterized, and its activity levels in normal mucosa and colonic adenocarcinoma have been determined. Sialidase activity was maximal at pH 5.5, and was unstable with storage at 4 and -20 degrees C. The bulk of activity was pellet-associated, and could not be released with triton X-100 or 3-([3-cholamidopropyl]- dimethylammonio)-1-propanesulfonate. Using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid as substrate, the Km and Vmax values were estimated to be 0.140 mmol/l and 63 mU/g, respectively. Furthermore, an inhibition by substrate concentrations above 1.5 mmol/l was detected. Neuraminic acid caused a competitive inhibition with a Ki of 3.5 mmol/l. A statistically significant increase (p < 0.001) in the sialidase specific activity was found in primary colonic adenocarcinoma (104.20 +/- 8.00 mU/g) compared to that of the normal mucosa (72.50 +/- 7.67 mU/g).