RESUMO
Essential hypertension (EH) is a chronic disease with clear epigenetic component. Inflammation and endothelial dysfunction have a great role in the development of persistent blood pressure elevation. The aim of this study was to explore an association between EH and DNA methylation in pro-inflammation cytokine gene interleukin-6 (IL-6) during the inflammatory process. We performed methylation analysis of peripheral blood DNA using bisulphite pyrosequencing technology between 96 EH patients and 96 age- and gender-matched healthy controls. The present results showed that three cytosine-phosphate-guanine (CpG) sites of IL-6 promoter CpG island had different lower methylation in EH group compared with controls, but only CpG2 (58.43±7.53 versus 62.34±9.65, P=0.004) and CpG3 (51.52±6.18 versus 57.45±8.29, P<0.001) had statistical difference. Logistic regression analysis found CpG3 hypomethylation was a risk factor of EH (odds ratio=1.111, adjusted P=0.004). In addition, we found hypermethylation of CpG1 (64.84±7.06 versus 61.84±8.61) and CpG2 (62.04±7.40 versus 59.30±9.57) in male compared with female. In male, we found hypomethylation of CpG2 (60.41±7.74 versus 64.28±6.36) and CpG3 (53.70±8.62 versus 57.78±7.87) of smoker versus non-smoker and hypomethylation of CpG2 (60.89±7.32 versus 64.70±7.03) and CpG3 (53.23±7.99 versus 60.48±7.58) of drinker versus non-drinker. Furthermore, the CpG2 and CpG3 had a negative correlation with systolic blood pressure and diastolic blood pressure (P<0.05). Receiver operating characteristic curve analysis showed that CpG2 (area under curve: 0.638, P=0.001) and CpG3 (area under curve: 0.704, P<0.001) had a diagnostic value to predict the risk of EH. In summary, our study revealed hypomethylation of IL-6 was correlated with the risk of EH in the population assessed and we found the differences of IL-6 promoter methylation in gender, smoking and drinking.
Assuntos
Pressão Sanguínea/genética , Metilação de DNA , Hipertensão Essencial/genética , Interleucina-6/genética , Fatores Etários , Consumo de Bebidas Alcoólicas/efeitos adversos , Área Sob a Curva , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Ilhas de CpG , Epigênese Genética , Hipertensão Essencial/diagnóstico , Hipertensão Essencial/fisiopatologia , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fenótipo , Regiões Promotoras Genéticas , Curva ROC , Fatores de Risco , Fatores Sexuais , Fumar/efeitos adversosAssuntos
Proteínas Fetais/metabolismo , Doença de Hodgkin/enzimologia , Doença de Hodgkin/patologia , Proteínas Tirosina Quinases/metabolismo , Adulto , Western Blotting , Medula Óssea/enzimologia , Medula Óssea/patologia , Proliferação de Células , Feminino , Proteínas Fetais/antagonistas & inibidores , Proteínas Fetais/genética , Doença de Hodgkin/genética , Humanos , Imunoprecipitação , Fosfopeptídeos/metabolismo , Fosforilação , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT5/genética , Fator de Transcrição STAT5/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tirosina/metabolismoAssuntos
Biomarcadores Tumorais/análise , Linhagem Celular Tumoral , Leucemia Mieloide/patologia , Proteínas de Neoplasias/metabolismo , Proteínas de Fusão Oncogênica/análise , Fosfotirosina/análise , Processamento de Proteína Pós-Traducional , Doença Aguda , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral/química , Cromossomos Humanos Par 12/genética , Cromossomos Humanos Par 12/ultraestrutura , Cromossomos Humanos Par 15/genética , Cromossomos Humanos Par 15/ultraestrutura , Humanos , Leucemia Mieloide/genética , Proteínas de Fusão Oncogênica/genética , Fosforilação , Estrutura Terciária de Proteína , Proteínas Tirosina Quinases/metabolismo , RNA Interferente Pequeno/farmacologia , Fator de Transcrição STAT5/metabolismo , Translocação GenéticaRESUMO
Core-binding factor alpha2 (CBFalpha2; otherwise known as AML1 or PEBP2alphaB) is a DNA-binding subunit in the family of core-binding factors (CBFs), heterodimeric transcription factors that play pivotal roles in multiple developmental processes in mammals, including hematopoiesis and bone development. The Runt domain in CBFalpha2 (amino acids 51 to 178) mediates DNA binding and heterodimerization with the non-DNA-binding CBFbeta subunit. Both the CBFbeta subunit and the DNA-binding protein Ets-1 stimulate DNA binding by the CBFalpha2 protein. Here we quantify and compare the extent of cooperativity between CBFalpha2, CBFbeta, and Ets-1. We also identify auto-inhibitory sequences within CBFalpha2 and sequences that modulate its interactions with CBFbeta and Ets-1. We show that sequences in the CBFalpha2 Runt domain and sequences C terminal to amino acid 214 inhibit DNA binding. Sequences C terminal to amino acid 214 also inhibit heterodimerization with the non-DNA-binding CBFbeta subunit, particularly heterodimerization off DNA. CBFbeta rescinds the intramolecular inhibition of CBFalpha2, stimulating DNA binding approximately 40-fold. In comparison, Ets-1 stimulates CBFalpha2 DNA binding 7- to 10-fold. Although the Runt domain alone is sufficient for heterodimerization with CBFbeta, sequences N terminal to amino acid 41 and between amino acids 190 and 214 are required for cooperative DNA binding with Ets-1. Cooperative DNA binding with Ets-1 is less pronounced with the CBFalpha2-CBFbeta heterodimer than with CBFalpha2 alone. These analyses demonstrate that CBFalpha2 is subject to both negative regulation by intramolecular interactions, and positive regulation by two alternative partnerships.
Assuntos
Proteínas de Ligação a DNA/genética , DNA/genética , Proteínas Proto-Oncogênicas/genética , Fatores de Transcrição/genética , Animais , Sítios de Ligação , Subunidade alfa 2 de Fator de Ligação ao Core , DNA/química , Proteínas de Ligação a DNA/química , Conformação de Ácido Nucleico , Ligação Proteica , Conformação Proteica , Proteína Proto-Oncogênica c-ets-1 , Proteínas Proto-Oncogênicas/química , Proteínas Proto-Oncogênicas c-ets , Fator de Transcrição AP-2 , Fatores de Transcrição/químicaRESUMO
Auto-inhibition is a common transcriptional control mechanism that is well characterized in the regulatory transcription factor Ets-1. Autoinhibition of Ets-1 DNA binding works through an inhibitory module that exists in two conformations. DNA binding requires a change in the inhibitory module from the packed to disrupted conformation. This structural switch provides a mechanism to tightly regulate Ets-1 DNA binding. We report that the Ets-1 partner protein core-binding factor alpha2 (CBFalpha2; also known as AML1 or PEBP2) stimulates Ets-1 DNA binding and counteracts auto-inhibition. Support for this conclusion came from three observations. First, the level of cooperative DNA binding (10-fold) was similar to the level of repression by auto-inhibition (10- to 20-fold). Next, a region necessary for cooperative DNA binding mapped to the inhibitory module. Third, an Ets-1 mutant with a constitutively disrupted inhibitory module did not bind DNA cooperatively with CBFalpha2. Furthermore, two additional lines of evidence indicated that CBFalpha2 affects the structural switch by direct interactions with Ets-1. First, the retention of cooperative DNA binding on nicked duplexes eliminated a potential role of through-DNA effects. Second, cooperative DNA binding was observed on composite sites with altered spacing or reversed orientation. We suggest that only protein interactions can accommodate this observed flexibility. These findings provide a mechanism by which CBF relieves the auto-inhibition of Ets-1 and illustrates one strategy for the synergistic activity of regulatory transcription factors.
Assuntos
Proteínas de Ligação a DNA , DNA/genética , Proteínas Proto-Oncogênicas/genética , Fatores de Transcrição/genética , Transcrição Gênica , Animais , Sítios de Ligação , Subunidade alfa 2 de Fator de Ligação ao Core , DNA/química , Regulação da Expressão Gênica , Conformação de Ácido Nucleico , Ligação Proteica , Conformação Proteica , Proteína Proto-Oncogênica c-ets-1 , Proteínas Proto-Oncogênicas/química , Proteínas Proto-Oncogênicas c-ets , Fatores de Transcrição/químicaRESUMO
Cbfa2 (AML1) encodes the DNA-binding subunit of a transcription factor in the small family of core-binding factors (CBFs). Cbfa2 is required for the differentiation of all definitive hematopoietic cells, but not for primitive erythropoiesis. Here we show that Cbfa2 is expressed in definitive hematopoietic progenitor cells, and in endothelial cells in sites from which these hematopoietic cells are thought to emerge. Endothelial cells expressing Cbfa2 are in the yolk sac, the vitelline and umbilical arteries, and in the ventral aspect of the dorsal aorta in the aorta/genital ridge/mesonephros (AGM) region. Endothelial cells lining the dorsal aspect of the aorta, and elsewhere in the embryo, do not express Cbfa2. Cbfa2 appears to be required for maintenance of Cbfa2 expression in the endothelium, and for the formation of intra-aortic hematopoietic clusters from the endothelium.
Assuntos
Aorta/embriologia , Proteínas de Ligação a DNA , Hematopoese/genética , Proteínas Proto-Oncogênicas , Fatores de Transcrição/genética , Animais , Antígenos de Superfície/imunologia , Antígenos de Superfície/metabolismo , Células da Medula Óssea , Diferenciação Celular , Subunidade alfa 2 de Fator de Ligação ao Core , Endotélio Vascular/embriologia , Citometria de Fluxo , Regulação da Expressão Gênica no Desenvolvimento , Marcação de Genes , Fígado/embriologia , Camundongos , Microscopia Eletrônica , Células-Tronco/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Consistent chromosomal rearrangements are found in a large number of hematopoietic tumors. In many cases, these rearrangements disrupt genes whose normal function is required for the proper development of blood cells. Excellent examples are the chromosomal rearrangements t(8;21)(q22;q22), t(12;21)(p13;q22), and inv(16)(p13q22) that disrupt two of the genes encoding a small family of heterodimeric transcription factors, core-binding factors (CBFs). CBFs consist of a DNA-binding CBFalpha subunit and a non-DNA-binding CBFbeta subunit. The t(8;21), associated with de novo acute myeloid leukemias, disrupts the CBFA2 (AML1) gene, which encodes a DNA-binding CBFalpha subunit. The t(12;21), the most common translocation in pediatric acute lymphocytic leukemias, also disrupts CBFA2. The CBFB gene, which encodes the non-DNA-binding subunit of the CBFs, is disrupted by the inv(16) in de novo acute myeloid leukemias. All chromosomal rearrangements involving the CBFA2 and CBFB genes create chimeric proteins, two of which have been unequivocally demonstrated to function as transdominant negative inhibitors of CBF function. Both the Cbfa2 and Cbfb genes are essential for normal hematopoiesis in mice, because homozygous disruption of either gene blocks definitive hematopoiesis. Recent data suggest that Cbfa2 and Cbfb are required for the emergence of definitive hematopoietic stem cells in the embryo from a putative definitive hemangioblast precursor. The transdominant negative inhibitor of CBF created by the inv(16), when present from the beginning of embryogenesis, also blocks the emergence of definitive hematopoietic cells in the embryo. On the other hand, chromosomal translocations involving the CBFA2 and CBFB genes in leukemias block hematopoiesis at later steps. This may reflect a difference in the timing at which translocations are acquired in the leukemias, which presumably is subsequent to emergence of the definitive hematopoietic stem cell. The cumulative data suggest that although the earliest requirement for Cbfa2 and Cbfb is for emergence of definitive hematopoietic stem cells, both genes are also required at later stages in the differentiation of some hematopoietic lineages.
Assuntos
Proteínas de Ligação a DNA/fisiologia , Hematopoese/fisiologia , Leucemia/fisiopatologia , Proteínas de Neoplasias , Fatores de Transcrição/fisiologia , Animais , Fatores de Ligação ao Core , Células-Tronco Hematopoéticas/fisiologia , Humanos , Leucemia Experimental/fisiopatologia , Translocação GenéticaRESUMO
The CBFbeta subunit is the non-DNA-binding subunit of the heterodimeric core-binding factor (CBF). CBFbeta associates with DNA-binding CBFalpha subunits and increases their affinity for DNA. Genes encoding the CBFbeta subunit (CBFB) and one of the CBFalpha subunits (CBFA2, otherwise known as AML1) are the most frequent targets of chromosomal translocations in acute leukemias in humans. We and others previously demonstrated that homozygous disruption of the mouse Cbfa2 (AML1) gene results in embryonic lethality at midgestation due to hemorrhaging in the central nervous system and blocks fetal liver hematopoiesis. Here we demonstrate that homozygous mutation of the Cbfb gene results in the same phenotype. Our results demonstrate that the CBFbeta subunit is required for CBFalpha2 function in vivo.
Assuntos
Sistema Nervoso Central/patologia , Proteínas de Ligação a DNA/genética , Genes Letais , Fígado/fisiopatologia , Proteínas Proto-Oncogênicas , Fatores de Transcrição/genética , Alelos , Animais , Células Sanguíneas/patologia , Sistema Nervoso Central/embriologia , Subunidade alfa 2 de Fator de Ligação ao Core , Subunidade beta de Fator de Ligação ao Core , Cruzamentos Genéticos , Proteínas de Ligação a DNA/metabolismo , Embrião de Mamíferos/patologia , Dosagem de Genes , Genótipo , Hematopoese/genética , Hemorragia/genética , Hibridização In Situ , Fígado/embriologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Mutagênese , Fenótipo , Reação em Cadeia da Polimerase , Ligação Proteica , RNA Mensageiro/isolamento & purificação , Células-Tronco , Fator de Transcrição AP-2 , Fatores de Transcrição/metabolismoRESUMO
The objective of this study was to determine if cytogenetic markers can be used as indicators of prior exposure to arsenic compounds. Baseline sister chromatid exchange (SCE) and mitomycin C-induced (MMC) SCE were measured in four study populations recruited from a blackfoot (BF) disease endemic area, including 22 patients with cancer (CA) only, 8 patients with both BF disease and CA (BF+CA), 10 patients with BF disease only, and 26 healthy residents (HRs). Another group of 23 healthy, nonarsenic-exposed workers were recruited as external healthy controls (HCs). Characteristics of study population were collected by questionnaire, and 10 ml of venous blood were drawn for lymphocyte culture. The results showed that the frequencies of baseline SCE did not differ among the five study groups. The frequencies of delta SCE (MMC-induced SCE minus baseline SCE) in CA only, BF disease only, and HRs, three arsenic-exposed groups, were significantly higher than in HCs. The frequency of delta SCE in the BF+CA group was nonsignificantly higher than in HCs, probably due to small sample size. The frequencies of both baseline SCE and delta SCE did not differ among CA only, BF disease only, BF+CA, and HR groups. The observation that baseline SCE did not increase in the arsenic-exposed populations indicates either that exposures were insufficiently high to change this marker or that lesions did not persist. The increased SCE response to MMC in arsenic-exposed populations suggests that previous arsenic exposure may result in hypersensitivity of human lymphocytes to carcinogens and/or mutagens. Both baseline SCE and delta SCE were not different among patients with arsenic-induced diseases and healthy normal residents, indicating that hypersensitivity may have been due to previous arsenic exposure but was not associated with disease status.
Assuntos
Arsênio/efeitos adversos , Exposição Ambiental , Mitomicina/farmacologia , Venenos/efeitos adversos , Troca de Cromátide Irmã/efeitos dos fármacos , Idoso , Biomarcadores , Carcinógenos/efeitos adversos , Estudos de Viabilidade , Feminino , Água Doce , Predisposição Genética para Doença , Humanos , Linfócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Mutagênicos/efeitos adversos , Neoplasias/genética , Doenças Vasculares Periféricas/induzido quimicamente , Doenças Vasculares Periféricas/genética , Transtornos da Pigmentação/induzido quimicamente , Transtornos da Pigmentação/genética , Fatores de Risco , Fumar/genética , TaiwanRESUMO
The purpose of this study was to evaluate the health hazards of firefighters after fighting a department store fire which lasted for 40 hours. Respiratory symptoms of 168 firefighters were collected and the pulmonary functions of 149 firefighters were measured by spirometer two days after fighting the fire and compared to 32 controls. The principal symptoms manifested by firefighters were burning eyes and mucous membrane irritation. Cross-sectional study of the pulmonary functions showed that FEV1.0, FEV1.0/FVC and the flow rates (MMF, FEF25%, and FEF75%) in smoking exposed firefighters were significantly lower than those in smoking controls. However, only FEV1.0 (%) was significantly lower in nonsmoking exposed firefighters than in nonsmoking controls. Both FVC and FEV1.0 showed a declining trend with the duration of fire fighting. This study provides further evidence that firefighters are exposed to irritants during fighting a fire. Exposure to the combustion products could lead to pulmonary function defects, especially in smoking firefighters.