RESUMO
Accumulating evidences suggest that glutamate plays a key role in the proliferation and invasion of malignant glioblastoma (GBM) tumors. It has been shown that GBM cells release and exploit glutamate for proliferation and invasion through AMPA glutamate receptors. Additionally, amplification of the epidermal growth factor receptor (EGFR) gene occurs in 40-50% of GBM. Since, PI3K/Akt is considered one of the main intracellular pathways involved in EGFR activation, AKT functions could trigger EGFR signaling. Thus, we investigated whether EGFR-phospho-Akt pathway is involved on the glutamate inducing U-87MG human GBM cell line proliferation. For these purpose, we treated the U-87MG cell line with 5 to 200 mM of glutamate and assessed the number of viable cells by trypan blue dye exclusion test. An increase in cell number (50%) was found at 5 mM glutamate, while the addition of DNQX (500 microM), an antagonist of AMPA receptor, inhibited the effect of glutamate on the U87-MG cells proliferation. Also, at 5 mM glutamate we observed an increase on the EGFR and phospho-Akt contents evaluated by immunohistochemistry. Moreover, U-87MG cells treated with glutamate exhibited an increase about 2 times in the EGFR mRNA expression. While, in the presence of the anti-EGFR gefitinib (50 muM) or the PI3K inhibitor wortmannin (5 muM), the U-87MG proliferation was restored to control levels. Together, our data suggest that glutamate signaling mediated by AMPA receptor induces U-87MG human GBM cell line proliferation via EGFR-phospho-Akt pathway.
Assuntos
Proliferação de Células/efeitos dos fármacos , Receptores ErbB/efeitos dos fármacos , Glioblastoma/metabolismo , Ácido Glutâmico/metabolismo , Linhagem Celular Tumoral , Receptores ErbB/metabolismo , Humanos , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de AMPA/efeitos dos fármacos , Receptores de AMPA/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologiaRESUMO
Biochemical markers of cellular stress/injury have been proposed to indicate outcome after head injury. The aim of the present study was to determine whether plasma von Willebrand factor (VWF) levels correlate with primary outcome and with clinical variables in severe traumatic brain injury (TBI). Forty-four male patients, victims of severe TBI, were analyzed. Clinical outcome variables of severe TBI comprised survival and neurological assessment using the Glasgow Outcome Scale (GOS) at intensive care unit (ICU) discharge. Computerized tomography (CT) scans were analyzed according to Marshall CT classification. Three consecutive venous blood samples were taken: first sample (11.4 +/- 5.2 h after trauma, mean +/- SD), and 24 h and 7 days later. The result of mean plasma VWF concentration was significantly higher in the TBI group (273 U/dL) than in the control group (107 U/dL; p < 0.001). Severe TBI was associated with a 50% mortality rate. Nonsurvivors presented significantly higher APACHE II scores than survivors (nonsurvivors mean, 18.8; survivors mean, 12.7; p < 0.001), and also presented higher scores in Marshall CT classification (nonsurvivors mean, 4.6; survivors mean, 2.7; p < 0.05). There was a significant positive correlation between plasma levels at second plasma sampling and scores in Marshall CT classification (p < 0.05). The sensitivity of plasma VWF concentration in predicting mortality according to the cut-off of 234 U/dL was 64%, with a specificity of 68%. Therefore, VWF increases following severe TBI may be a marker of unfavorable outcome.
Assuntos
Lesões Encefálicas/sangue , Lesões Encefálicas/mortalidade , Fator de von Willebrand/metabolismo , Adolescente , Adulto , Biomarcadores/sangue , Lesões Encefálicas/diagnóstico por imagem , Seguimentos , Escala de Resultado de Glasgow , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Radiografia , Taxa de Sobrevida , Resultado do TratamentoRESUMO
OBJECTIVES: Severe traumatic brain injury (TBI) is associated with a 30-70% mortality rate. Nevertheless, controversy has been raised concerning the prognostic value of biomarkers following severe TBI. Therefore, our aim was to determine whether sFas or TNFalpha serum levels correlate with primary outcome following isolated severe TBI. METHODS: Seventeen consecutive male patients, victims of isolated severe TBI (Glasgow Coma Scale score 3-8) and a control group consisting of 6 healthy male volunteers were enrolled in this prospective study. Clinical outcome variables of severe TBI comprised: survival, time for intensive care unit (ICU) discharge, and neurological assessment by Glasgow Outcome Scale at ICU discharge. Venous blood samples were taken at admission in the ICU. Serum sFas and TNFalpha concentrations were measured by ELISA assays. RESULTS: At admission in the ICU (mean time 10.2 h after injury), mean sFas and TNFalpha concentrations were significantly increased in the TBI (0.105 and 24.275 rhog/l, respectively) compared with the control group (0.047 and 15.475 rhog/l, respectively). However, no significant correlation was found between higher serum sFas or TNFalpha concentrations and fatal outcome. CONCLUSIONS: Increased serum sFas and TNFalpha levels following isolated severe TBI did not predict fatal outcome.
Assuntos
Lesões Encefálicas/sangue , Lesões Encefálicas/mortalidade , Fator de Necrose Tumoral alfa/sangue , Receptor fas/sangue , Adulto , Estudos de Casos e Controles , Seguimentos , Escala de Coma de Glasgow , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Taxa de SobrevidaRESUMO
BACKGROUND AND AIMS: Colorectal cancer is a common disease with high rate of mortality. Although there is evidence of some benefits of 5-fluorouracil (5-FU), the most commonly used drug in colon cancer therapy, it still remains unsatisfactory because of intrinsic or acquired drug resistance. Heat shock proteins (Hsps) synthesis can be increased by cellular insults, such as chemotherapy-induced damage. Inducible Hsp70 has been suggested to be involved in cytoprotection against apoptosis. In the present study, we investigated whether the content of Hsp70 is associated to 5-FU resistance. METHODS: HT-29 and SNU-C4 human colon cancer cell lines were treated with 5-FU and their relative chemoresistance, and Hsp70 were determined. RESULTS: Comparison of IC(50) values showed that the HT-29 cells were relatively resistant to 5-FU, whereas the SNU-C4 cells presented greater sensitivity to this drug. Further, 5-FU treatment leads to a hypodiploid population in HT-29 cells significantly lower compared to SNU-C4 cells. In the HT-29 cell line, 5-FU treatment promoted an increase of 5.5 times in Hsp70 concentration after 12 h. Then, within 24 h, the increase in Hsp70 levels was still about two times. In contrast, in the SNU-C4 cell line, 5-FU induced an increase of about two times in the Hsp70 content after 12 h and, after 24 h, did not significantly affect Hsp70 content. CONCLUSIONS: These data suggest that 5-FU induced Hsp70 synthesis in the HT-29 resistant cell line and that this Hsp70 accumulation could protect against 5-FU-induced apoptosis. Thus, Hsp70 protection against 5-FU-induced apoptosis might underlie colon cancer chemoresistance.
Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/metabolismo , Fluoruracila/farmacologia , Proteínas de Choque Térmico HSP70/metabolismo , Antineoplásicos/administração & dosagem , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Fluoruracila/administração & dosagem , Humanos , Fatores de TempoRESUMO
BACKGROUND: Severe traumatic brain injury (TBI) is associated with a 30%-70% mortality rate. S100B has been proposed as a biomarker for indicating outcome after TBI. Nevertheless, controversy has arisen concerning the predictive value of S100B for severe TBI in the context of multitrauma. Therefore, our aim was to determine whether S100B serum levels correlate with primary outcome following isolated severe TBI or multitrauma in males. METHODS: Twenty-three consecutive male patients (age 18-65 years), victims of severe TBI [Glasgow Coma Scale (GCS) 3-8] (10 isolated TBI and 13 multitrauma with TBI) and a control group consisting of eight healthy volunteers were enrolled in this prospective study. Clinical outcome variables of severe TBI comprised: survival, time to intensive care unit (ICU) discharge, and neurological assessment [Glasgow Outcome Scale (GOS) at ICU discharge]. Venous blood samples were taken at admission in the ICU (study entry), 24 h later, and 7 days later. Serum S100B concentration was measured by an immunoluminometric assay. RESULTS: At study entry (mean time 10.9 h after injury), mean S100B concentrations were significantly increased in the patient with TBI (1.448 microg/L) compared with the control group (0.037 microg/L) and patients with fatal outcome had higher mean S100B (2.10 microg/L) concentrations when compared with survivors (0.85 microg/L). In fact, there was a significant correlation between higher initial S100B concentrations and fatal outcome (Spearman's =0.485, p=0.019). However, there was no correlation between higher S100B concentrations and the presence of multitrauma. The specificity of S100B in predicting mortality according to the cut-off of 0.79 microg/L was 73% at study entry. CONCLUSIONS: Increased serum S100B levels constitute a valid predictor of unfavourable outcome in severe TBI, regardless of the presence of associated multitrauma.
Assuntos
Lesões Encefálicas/mortalidade , Lesões Encefálicas/fisiopatologia , Fatores de Crescimento Neural/sangue , Proteínas S100/sangue , Adulto , Biomarcadores/sangue , Brasil/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Curva ROC , Subunidade beta da Proteína Ligante de Cálcio S100 , Análise de SobrevidaRESUMO
Severe traumatic brain injury (TBI) is associated with a 35-70% mortality rate. Biochemical markers of cellular stress/injury have been proposed to indicate outcome after head injury. Therefore, our aim was to determine whether Hsp70 could be detected in the serum of patients after severe TBI and whether serum levels of Hsp70 correlate with primary outcome in severe TBI. Twenty consecutive male patients, victims of severe TBI (GCS 3-8), were enrolled in this prospective study. Clinical outcome variables of severe TBI comprised: survival, time for ICU discharge, and neurological assessment using the Glasgow Outcome Scale (GOS) at the ICU discharge. Venous blood samples were taken at admission in the ICU (study entry), 24 h later, and 7 days later. A control group consisting of eight healthy male volunteers was also included. Serum Hsp70 levels were measured by an enzyme-linked immunosorbent assay. Mean serum Hsp70 concentrations were significantly increased in the TBI (97.6, 48.1, and 39.2 ng/mL, at study entry, 24 h later, and 7 days later, respectively) compared with the control group (12.2 ng/mL). Severe TBI was associated with a 50% mortality rate. On study entry (mean time 10.8 h after injury), a higher proportion of patients with fatal outcome had elevated serum Hsp70 (mean 143.5 ng/mL) concentrations when compared with survivors (mean 51.6 ng/mL). There was a significant correlation between higher initial serum Hsp70 concentrations and fatal outcome. The sensitivity of serum Hsp70 predicting mortality according to the cutoff of 62 ng/mL is 70% within 20 h after injury. Increased serum Hsp70 levels may constitute an early predictor of unfavorable outcome in severe TBI in males.
Assuntos
Lesões Encefálicas/sangue , Lesões Encefálicas/mortalidade , Proteínas de Choque Térmico HSP70/sangue , APACHE , Adolescente , Adulto , Biomarcadores/sangue , Ensaio de Imunoadsorção Enzimática , Escala de Resultado de Glasgow , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Curva ROC , Sensibilidade e Especificidade , Taxa de SobrevidaRESUMO
Anticancer agents may be derived either from the isolation of an active lead compound occurring spontaneously in nature or by novel chemical synthesis in the laboratory. There are examples of successful drugs being derived from both sources, which have had a profound impact on the natural history of various types of cancer. The treatment of lymphomas and acute leukaemias with the use of combination chemotherapy, including anthracyclines and vinca alkaloids, are examples of the contribution of nature. In contrast, agents such as 5-fluorouracil, methotrexate and more recently, the humanised anti-CD20 antibody rituximab and the tyrosine kinase inhibitor imatinib are examples of synthetic compounds, which were designed with a clear rationale, that are routinely used in patients with solid tumours and haematological malignancies. Until recently, the tradition in natural product-derived anticancer drug development was to rely almost exclusively on the screening of terrestrial sources (plant extracts and fermentation products) for their cytotoxic properties. Although C-nucleosides obtained from Caribbean sponge were the initial inspiration for the synthesis of antiviral substituted nucleosides and the successful anticancer agent citarabine, active against leukaemias and lymphomas, the contribution of marine compounds as a source of anticancer agents was modest. In recent years, the improvements in the technology of deep-sea collection and aquaculture added to the growing recognition of the tremendous biodiversity present in the marine world, and has contributed to the growing interest of exploring the oceans as a potential source of new anticancer candidates. This is reflected in the number of marine-derived compounds undergoing preclinical and early clinical development. In this paper, the authors discuss the available literature on anticancer agents that have reached clinical trials, such as didemnin B, aplidine, dolastatin-10, bryostatin-1 and ecteinascidin-743 (ET-743, trabectedin), as well as other promising compounds still undergoing tests in the laboratory.