RESUMO
Extracellular vesicles (EVs) are considered as important mediators of intercellular communication, which carry a diverse repertoire of genetic information between cells. This feature of EVs can be used and improved to advance their therapeutic potential. We have previously shown that genetically engineered EVs carrying the suicide gene mRNA and protein-cytosine deaminase (CD) fused to uracil phosphoribosyltransferase (UPRT)-inhibited schwannoma tumor growth in vivo. To further examine whether this approach can be applied to other cancer types, we established a subcutaneous xenograft glioblastoma tumor model in mice, as glioblastoma represents the most common primary brain tumor, which is highly aggressive compared with the original schwannoma tumor model. U87-MG glioblastoma cells were implanted into the flanks of nude SCID mice, and the animals were intratumorally injected with the EVs isolated from the cells expressing EGFP or CD-UPRT. After the intraperitoneal administration of the prodrug 5-fluorocytosine, the tumor growth was assessed by regular caliper measurements. Our data revealed that the treatment with the CD-UPRT-enriched EVs significantly reduced the tumor growth in mice. Taken together, our findings suggest that EVs uploaded with therapeutic CD-UPRT mRNA/protein may be a useful tool for glioblastoma treatment.
Assuntos
Sistemas de Liberação de Medicamentos , Vesículas Extracelulares/metabolismo , Técnicas de Transferência de Genes , Genes Transgênicos Suicidas , Glioblastoma/genética , Glioblastoma/metabolismo , RNA Mensageiro , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Transporte Biológico , Ciclo Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Modelos Animais de Doenças , Feminino , Expressão Gênica , Engenharia Genética , Glioblastoma/patologia , Glioblastoma/terapia , Humanos , Camundongos , Camundongos SCID , Pentosiltransferases/genética , Pentosiltransferases/metabolismo , Pró-Fármacos/farmacologia , Carga Tumoral/efeitos dos fármacos , Carga Tumoral/genética , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
miR-200a has been implicated in the pathogenesis of meningiomas, one of the most common central nervous system tumors in humans. To identify how miR-200a contributes to meningioma pathogenesis at the molecular level, we used a comparative protein profiling approach using Gel-nanoLC-MS/MS and identified approximately 130 dysregulated proteins in miR-200a-overexpressing meningioma cells. Following the bioinformatic analysis to identify potential genes targeted by miR-200a, we focused on the non-muscle heavy chain IIb (NMHCIIb), and showed that miR-200a directly targeted NMHCIIb. Considering the key roles of NMHCIIb in cell division and cell migration, we aimed to identify whether miR-200a regulated these processes through NMHCIIb. We found that NMHCIIb overexpression partially rescued miR-200a-mediated inhibition of cell migration, as well as cell growth in vitro and in vivo. Moreover, siRNA-mediated silencing of NMHCIIb expression resulted in a similar migration phenotype in these cells and inhibited meningioma tumor growth in mice. Taken together, these results suggest that NMHCIIb might serve as a novel therapeutic target in meningiomas.
Assuntos
Neoplasias Meníngeas/patologia , Meningioma/patologia , MicroRNAs/genética , Cadeias Pesadas de Miosina/genética , Miosina não Muscular Tipo IIB/genética , Animais , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células/genética , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Meníngeas/genética , Meningioma/genética , Camundongos , Camundongos Nus , Cadeias Pesadas de Miosina/antagonistas & inibidores , Cadeias Pesadas de Miosina/biossíntese , Transplante de Neoplasias , Miosina não Muscular Tipo IIB/antagonistas & inibidores , Miosina não Muscular Tipo IIB/biossíntese , Interferência de RNA , RNA Interferente Pequeno , Transplante HeterólogoRESUMO
Minichromosome maintenance (MCM) proteins are key elements that function as a part of the pre-replication complex to initiate DNA replication in eukaryotes. Consistent with their roles in initiating DNA replication, overexpression of MCM family members has been observed in several malignancies. Through bioinformatic analysis of The Cancer Genome Atlas's data on glioblastoma multiforme (GBM), we found that the genomic region containing MCM7 gene was amplified in more than 80% of the present cases. To validate this finding and to identify the possible contribution of the remaining members of the MCM family to GBM progression, we used quantitative real-time PCR to analyze the gene expression profiles of all MCM family members in Grade IV (GBM) tissue samples and observed a significant upregulation in GBM samples compared with normal white matter tissues. In addition, we compared the observed gene expression profiles with those of Grade II and Grade III astrocytoma samples and determined that the observed upregulation was restricted and specific to Grade IV. MCM7 was the most upregulated gene in the gene set we analyzed, and therefore we wanted to identify the role of MCM7 in GBM progression. We determined that siRNA-mediated knockdown of MCM7 expression reduced GBM cell proliferation and also inhibited tumor growth in both xenograft and orthotopic mouse models of GBM. Taken together, our data suggest that MCM7 can be a potential prognostic marker and a novel therapeutic target in GBM therapy.
Assuntos
Neoplasias Encefálicas/metabolismo , Glioblastoma/metabolismo , Componente 7 do Complexo de Manutenção de Minicromossomo/genética , Animais , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Expressão Gênica , Técnicas de Silenciamento de Genes , Glioblastoma/patologia , Camundongos , Camundongos Nus , Componente 7 do Complexo de Manutenção de Minicromossomo/metabolismo , Transplante de Neoplasias , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , Carga TumoralRESUMO
Standard treatment for glioblastoma multiforme and other brain tumors consists of surgical resection followed by combined radio-/chemotherapy. However, radiation resistance of tumor cells limits the success of this treatment, and the tumors invariably recur. Therefore, the selective inhibition of molecular mediators of radiation resistance may provide therapeutic benefit to the patient. One of these targets is the Rad51 protein, which is a key component of the homologous recombinational repair of DNA double-strand breaks. Here, we investigated whether post-transcriptional silencing of Rad51 by herpes simplex virus-type 1 (HSV-1) amplicon vector-mediated short interfering RNA expression can enhance the antitumor effect of radiation therapy. We demonstrate that these vectors specifically and efficiently inhibited the radiation-induced recruitment of Rad51 into nuclear foci in human glioma cells. The combination of vector-mediated silencing of Rad51 expression and treatment with ionizing radiation resulted in a pronounced reduction of the survival of human glioma cells in culture. In athymyc mice, a single intratumoral injection of Rad51-specific HSV-1 amplicon vector followed by a single radiation treatment resulted in a significant decrease in tumor size. In control animals, including mice that received an intratumoral injection of Rad51-specific amplicon vector but no radiation treatment, the tumor sizes increased.
Assuntos
Neoplasias Encefálicas/terapia , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Glioma/terapia , Herpesvirus Humano 1/genética , Rad51 Recombinase/antagonistas & inibidores , Animais , Apoptose , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/radioterapia , Linhagem Celular Tumoral , Terapia Combinada , Feminino , Expressão Gênica , Inativação Gênica , Vetores Genéticos/genética , Glioblastoma/terapia , Glioma/patologia , Glioma/radioterapia , Humanos , Injeções Intralesionais , Camundongos , Camundongos Nus , Interferência de RNA , RNA Interferente Pequeno/genética , Rad51 Recombinase/genética , Tolerância a Radiação/fisiologia , Resultado do TratamentoRESUMO
Atherosclerosis in childhood has a slowly progressive course and its clinical features usually become prominent in middle ages. Hypercholesterolemia is one of the major risk factors for the development of atherosclerosis. A clear correlation exists between hypercholesterolemia in childhood and atherosclerotic lesions extending into adulthood.In this study, we evaluated the effect of slow release theophylline (SRT) treatment on plasma lipid profile and assessed the risk for atherosclerotic coronary heart disease in children with bronchial asthma. Group 1 consisted of 15 children with a mean age of 10.8 3.19 years who received SRT for bronchial asthma for a mean period of 9.13 2.17 months. Group 2 was composed of 15 children with a mean age of 11.40 3.78 years and followed up for bronchial asthma, who received no SRT treatment. Group 3 comprised 15 children with a mean age of 9.00 3.76 years and no history of asthma or wheezing. In all patients lipid profiles were assessed by measuring levels of plasma triglyceride, total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) apolipoprotein A (Apo-A) and apolipoprotein B (Apo-B). In group 1, the mean total cholesterol level was 175.53 24.36 mg/dl, LDL-C level was 91.00 24.07 mg/dl and Apo-B level was 87.27 12.74 mg/dl after SRT treatment. In group 1, group 2 (control group with asthma) and group 3 (the non-asthmatic control group), the mean plasma lipid level after SRT treatment was significantly higher than that before SRT treatment. In conclusion, long-term SRT treatment in children with bronchial asthma may alter lipid profile and may increase the risk for developing atherosclerotic coronary heart disease.
Assuntos
Antiasmáticos/efeitos adversos , Asma/sangue , Asma/tratamento farmacológico , Lipoproteínas/sangue , Teofilina/efeitos adversos , Antiasmáticos/administração & dosagem , Criança , Preparações de Ação Retardada , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Teofilina/administração & dosagemRESUMO
Atherosclerosis in childhood has a slowly progressive course and its clinical features usually become prominent in middle ages. Hypercholesterolemia is one of the major risk factors for the development of atherosclerosis. A clear correlation exists between hypercholesterolemia in childhood and atherosclerotic lesions extending into adulthood. In this study, we evaluated the effect of slow release theophylline (SRT) treatment on plasma lipid profile and assessed the risk for atherosclerotic coronary heart disease in children with bronchial asthma. Group 1 consisted of 15 children with a mean age of 10.8 ± 3.19 years who received SRT for bronchial asthma for a mean period of 9.13 ± 2.17 months. Group 2 was composed of 15 children with a mean age of 11.40 ± 3.78 years and followed up for bronchial asthma, who received no SRT treatment. Group 3 comprised 15 children with a mean age of 9.00 ± 3.76 years and no history of asthma or wheezing. In all patients lipid profiles were assessed by measuring levels of plasma triglyceride, total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) apolipoprotein A (Apo-A) and apolipoprotein B (Apo-B). In group 1, the mean total cholesterol level was 175.53 ± 24.36 mg/dl, LDL-C level was 91.00 ± 24.07 mg/dl and Apo-B level was 87.27 ± 12.74 mg/dl after SRT treatment. In group 1, group 2 (control group with asthma) and group 3 (the non-asthmatic control group), the mean plasma lipid level after SRT treatment was significantly higher than that before SRT treatment. In conclusion, long-term SRT treatment in children with bronchial asthma may alter lipid profile and may increase the risk for developing atherosclerotic coronary heart disease (AU)
La arteriosclerosis en la infancia experimenta un desarrollo progresivo lento y su descubrimiento clínico se suele producir en la edad madura. La hipercolesterolemia es uno de los mayores factores de riesgo en el desarrollo de arteriosclerosis. Existe una relación definida entre la hipercolesterolemia en la infancia y la prolongación de las lesiones arterioscleróticas en la edad adulta. En este estudio hemos evaluado el efecto de un tratamiento con teofilina de liberación prolongada en un perfil de lípido plasmático y hemos descrito el riesgo de arteriosclerosis coronaria en niños con asma bronquial. Quince niños con una edad media de 10,8 ± 3,19 años y que recibieron TLP durante un tiempo medio de 9,13 ± 2,17 meses contra el asma bronquial fueron clasificados como grupo 1; 15 niños con una edad media de 11,40 ± 3,78, expuestos a un seguimiento por asma bronquial y que no recibieron el tratamiento con TPL fueron clasificados como grupo 2; 15 niños con una edad media de 9,00 ± 3,76 años que no presentaban un historial de asma ni de disnea sibilante fueron clasificados como grupo 3. Los perfiles de lípidos se cuantificaron con mediciones de los niveles de triglicéridos en plasma, colesterol total, lipoproteínas de alta densidad (LAD), lipoproteínas de baja densidad (LBD), apolipoproteína A (Apo-A) y apolipoproteína B (Apo-B) a todos los pacientes objeto de este estudio. El nivel medio de colesterol total se determinó en 175,53 ± 24,36 mg/dl, el nivel de LBD en 91,00 ± 24,07 mg/dl y el nivel Apo-B en 87,27 ± 12,74 mg/dl en el primer grupo tras el tratamiento con TPL. El nivel medio de lípidos en plasma tras el tratamiento con TLP fue significativamente más alto que el nivel medio de lípidos antes del tratamiento en el Grupo 1, en el Grupo 2 (grupo de control con asma) y en el Grupo 3 (grupo de control sin asma). En conclusión, el tratamiento con TLP a largo plazo en niños con asma bronquial puede alterar el perfil de lípidos y puede incrementar el riesgo de padecer cardiopatías coronarias por arteriosclerosis en los niños con asma bronquial (AU)
Assuntos
Humanos , Masculino , Feminino , Criança , Teofilina/farmacocinética , Asma/tratamento farmacológico , Aterosclerose/epidemiologia , Hipercolesterolemia/epidemiologia , Preparações de Ação Retardada/farmacocinética , Lipoproteínas/análiseRESUMO
The metal-regulatory transcription factor 1 (MTF-1) is a key regulator of heavy metal-induced transcription of metallothionein I and II and other genes in mammals and other metazoans. Transcriptional activation of genes by MTF-1 is mediated through binding to metal-responsive elements of consensus TGCRCNC in the target gene promoters. In an attempt to further clarify the mechanisms by which certain external signals activate MTF-1 and in turn modulate gene transcription, we show here that human MTF-1 has a dual nuclear and cytoplasmic localization in response to diverse stress stimuli. MTF-1 contains a consensus nuclear localization signal located just N-terminal to the first zinc finger that contributes to but is not essential for nuclear import. MTF-1 also harbors a leucine-rich, nuclear export signal. Under resting conditions, the nuclear export signal is required for cytoplasmic localization of MTF-1 as indicated by mutational analysis and transfer to the heterologous green fluorescent protein. Export from the nucleus was inhibited by leptomycin B, suggesting the involvement of the nuclear export protein CRM1. Our results further show that in addition to the heavy metals zinc and cadmium, heat shock, hydrogen peroxide, low extracellular pH (pH 6.0), inhibition of protein synthesis by cycloheximide, and serum induce nuclear accumulation of MTF-1. However, heavy metals alone (and not the other stress conditions) induce a significant transcriptional response via metal-responsive element promoter sequences, implying that nuclear import of MTF-1 is necessary but not sufficient for transcriptional activation. Possible roles for nuclear import under non-metal stress conditions are discussed.
Assuntos
Fatores de Transcrição/fisiologia , Cádmio/farmacologia , Sequência Consenso , Cicloeximida/farmacologia , Citoplasma/metabolismo , Proteínas de Ligação a DNA , Ácidos Graxos Insaturados/farmacologia , Técnica Indireta de Fluorescência para Anticorpo , Temperatura Alta , Humanos , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Estresse Oxidativo , Regiões Promotoras Genéticas , Inibidores da Síntese de Proteínas/farmacologia , Ativação Transcricional , Transfecção , Células Tumorais Cultivadas , Zinco/farmacologia , Fator MTF-1 de TranscriçãoRESUMO
In this experimental study, we evaluated the effect of trimetazidine (TMZ) on renal ischemia-reperfusion (IR) injury in Sprague-Dawley rats. Renal IR was achieved by a 75-min clamping of the left renal pedicle and subsequent 24 h reperfusion, after right nephrectomy was performed. The rats were randomly divided into three groups: group 1 (sham-operated: no IR injury), group 2 (ischemic control: saline treatment), and group 3 (3 mg/kg TMZ before ischemia). After 24 h of reperfusion, blood samples and renal tissue samples were taken to measure the levels of creatinine, tissue malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) activity. Histopathological changes were evaluated. In addition, the 7-day survival rates in each group were evaluated. We found significant increases in the levels of creatinine and tissue MDA, severe acute tubular necrosis, and a significant decrease in the activity of the GSH-Px in group 2. There were significant decreases in the levels of creatinine and tissue MDA, mild acute tubular necrosis, and a significant increase in activity of the GSH-Px in group 3 when compared with the control group (p <0.05). Statistically significant differences (p <0.05) in survival were noted between the ischemic control and sham-operated and TMZ groups. We have concluded that TMZ is able to protect the kidney from warm IR injury.
Assuntos
Isquemia/tratamento farmacológico , Rim/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Trimetazidina/farmacologia , Vasodilatadores/farmacologia , Animais , Glutationa/metabolismo , Isquemia/metabolismo , Isquemia/patologia , Rim/metabolismo , Rim/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
This study was performed to elucidate the lung glutathione-related defense potential in tumoral tissues. Reduced (GSH) and oxidized (GSSG) glutathione, glutathione reductase (GR), selenium-dependent (SeGPx) and total glutathione peroxidase (tGPx), and glutathione S-transferase (GST) activities in 38 tumoral lung tissues and 17 normal lung tissues were determined to obtain a comprehensive profile of the lung glutathione and glutathione-related enzymes in cancer. The enzyme levels in tumoral tissues (n = 38) were found to be significantly higher (P < 0.05) than those in normal tissues (n = 17). Reduced glutathione levels, and not oxidized glutathione levels, were found to be higher in normal tissues than those in tumoral tissues. We found no statistically significant difference between the adenocarcinoma and squamous cell carcinoma groups for any of the parameters studied.
Assuntos
Glutationa Peroxidase/análise , Glutationa Redutase/análise , Glutationa Transferase/análise , Glutationa/análise , Neoplasias Pulmonares/química , HumanosRESUMO
BACKGROUND: The metabolic and hemodynamic effects of nisoldipine supplementation in cardioplegia after ischemic injury were investigated in 13 isolated rabbit hearts. Group 1 consisted of 6 hearts, which received St. Thomas II cardioplegic solution. In group 2, nisoldipine was added to the cardioplegic solution at a concentration of 0.1 mg/kg in 7 hearts. METHODS: The explanted hearts were suspended from Langendorff apparatus and were perfused with Krebs-Henseleit solution. Left ventricular pressure, heart rate, malondialdehyde, glutathione peroxidase, glutathione reductase, reduced glutathione, oxidized glutathione, creatine kinase MB, (CK-MB), aspartate transaminase, and lactate dehydrogenase (LDH) were measured before and after 60 minutes of ischemia. Peak generated pressure after ischemia was significantly higher in group 2 versus group 1 while end-diastolic pressure was significantly lower in group 2 after ischemic arrest (P <.05). RESULTS: Malondialdehyde levels were lower in group 2 (P <.05). Glutathione peroxidase and glutathione reductase levels were significantly higher in group 2 (P <.05). The only enzymatic significant difference was observed between the preischemic and postischemic levels of aspartate transaminase in group 2 (P <.05). CONCLUSIONS: These findings show beneficial effects of nisoldipine cardioplegia, although its use as a cardioplegic additive is not yet possible. We believe, however, the effects of oral nisoldipine before cardiac surgery can be studied in a clinical setting.
