RESUMEN
G-quadruplexes are non-canonical DNA secondary structures involved in several genomic and molecular processes. Here, we summarize the main G-quadruplex features and evidences proving the in vivo role on the transcriptional regulation of genes required for zebrafish embryonic development. We also discuss alternative strategies for specifically interfering G-quadruplex in vivo.
Asunto(s)
ADN/química , Transcripción Genética , Pez Cebra/embriología , Animales , G-Cuádruplex , Regulación del Desarrollo de la Expresión Génica , Modelos Moleculares , Conformación de Ácido Nucleico , Pez Cebra/genéticaRESUMEN
Although BMP4-induced differentiation of glioma stem cells (GSCs) is well recognized, details of the cellular responses triggered by this morphogen are still poorly defined. In this study, we established several GSC-enriched cell lines (GSC-ECLs) from high-grade gliomas. The expansion of these cells as adherent monolayers, and not as floating neurospheres, enabled a thorough study of the phenotypic changes that occurred during their differentiation. Herein, we evaluated GSC-ECLs' behavior toward differentiating conditions by depriving them of growth factors and/or by adding BMP4 at different concentrations. After analyzing cellular morphology, proliferation and lineage marker expression, we determined that GSC-ECLs have distinct preferences in lineage choice, where some of them showed an astrocyte fate commitment and others a neuronal one. We found that this election seems to be dictated by the expression pattern of BMP signaling components present in each GSC-ECL. Additionally, treatment of GSC-ECLs with the BMP antagonist, Noggin, also led to evident phenotypic changes. Interestingly, under certain conditions, some GSC-ECLs adopted an unexpected smooth muscle-like phenotype. As a whole, our findings illustrate the wide differentiation potential of GSCs, highlighting their molecular complexity and paving a way to facilitate personalized differentiating therapies.
Asunto(s)
Proteína Morfogenética Ósea 4/metabolismo , Neoplasias Encefálicas/patología , Proteínas Portadoras/metabolismo , Glioma/patología , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Anciano , Antígenos CD/metabolismo , Proteína Morfogenética Ósea 4/farmacología , Proteínas Portadoras/farmacología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Péptidos y Proteínas de Señalización Intercelular/farmacología , Masculino , Persona de Mediana Edad , Proteínas del Tejido Nervioso/metabolismo , Fenotipo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Células Tumorales Cultivadas/patología , Células Tumorales Cultivadas/fisiologíaRESUMEN
The Mexican tetra (Astyanax mexicanus), a freshwater teleost fish, is an excellent vertebrate model organism to study tooth development, specifically the spatiotemporal events related to the development of the oral and pharyngeal dentitions. In contrast to the coordinated early tooth development in the premaxilla and mandible, the maxillary teeth develop much later in life at 60 dpf. By analysing a growth series of bone and cartilage stained tetra and histological sectioning of the tooth bearing bones, we track the developmental events of tooth development over ontogeny of this animal. Whole mount in situ hybridisation with bone morphogenetic proteins and their inhibitor Noggin was conducted to track the late tooth development events. Our data show that the first generation teeth are small and unicuspid irrespective of their location. Oral jaw teeth become multicuspid and large over ontogeny while the pharyngeal dentition remains unicuspid and disorganised. Tooth eruption occurs late in the maxillary bone. The distinct expression pattern of the BMP antagonist, Noggin, suggests that Noggin plays an inhibitory role by preventing early tooth development in the maxillary bone. These data further support and highlight the use of the Mexican tetra in understanding the spatio-temporal differences in tooth development in vertebrate jaws.
Asunto(s)
Characidae/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Desarrollo Maxilofacial/fisiología , Diente/crecimiento & desarrollo , Animales , Proteína Morfogenética Ósea 4/genética , Proteína Morfogenética Ósea 4/metabolismo , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismoRESUMEN
O carcinoma epidermóide oral (CEO) representa a neoplasia maligna mais prevalente na cavidade oral e por atingir um grande número de indivíduos, acaba se tornado um relevante problema de saúde pública. Muitos estudos demonstram alterações nos componentes da via BMP em vários tipos de tumores, como os de próstata, cólon, mama, gástricos e CEOs. É do conhecimento atual que essas proteínas podem exercer efeito pró-tumoral em estágios mais avançados do desenvolvimento neoplásico vindo a favorecer a progressão e invasão tumoral. A inibição da via de sinalização da BMP-2, através dos seus antagonistas, tem mostrado resultados positivos de ação antitumoral e que assim, o uso do Noggin pode ser um novo alvo terapêutico contra o câncer. Diante destas evidências e dos escassos trabalhos com BMP-2, Noggin e CEO, o objetivo desta pesquisa foi avaliar o efeito da BMP-2 e seu antagonista Noggin sobre a proliferação e migração celulares em culturas de células de carcinoma epidermóide de língua humana (SCC25). Foi feita a divisão em três grupos de estudo, um grupo controle, onde as células SCC25 não sofriam tratamento com substância alguma, um grupo BMP-2, no qual as células eram tratadas com 100ng/ml de BMP-2 e um grupo de células que eram tratadas com 100ng/ml de Noggin. Para o ensaio de proliferação e ciclo celular foram estabelecidos três intervalos de tempo (24, 48 e 72 horas). A atividade proliferativa foi investigada por azul de tripan e a análise do ciclo celular através da marcação por iodeto de propídio em Citometria de fluxo. O potencial de migração/invasão das células SCC25 foi avaliado através da realização de um ensaio de invasão celular utilizando o matrigel em um intervalo de 48 horas. A curva de proliferação revelou maior crescimento celular nas células tratadas com BMP-2 no intervalo de 72 horas (p<0.05) e menor crecimento e viabilidade celular no grupo Noggin. As proteínas recombinantes favoreceram a maior porcentagem das células na fase do ciclo celular Go/G1 com diferença estatisticamente significativa no intervalo de 24 horas (p<0,05). A BMP-2 promoveu uma maior invasão das células estudadas, assim como o seu antagonista Noggin inibiu a invasão das células estudadas (p<0,05). Dessa forma, os resultados indicam que a BMP-2 favorece o fenótipo maligno, pois estimula a proliferação e invasão das células SCC25 e seu antagonista Noggin pode ser uma alternativa terapêutica pois inibiu essas características pró-tumorais. (AU)
Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression. (AU)
Asunto(s)
Carcinoma de Células Escamosas/fisiopatología , Neoplasias de la Lengua/fisiopatología , Proliferación Celular , Citometría de Flujo/métodos , Estadísticas no Paramétricas , Técnicas In Vitro , Metástasis de la NeoplasiaRESUMEN
O carcinoma epidermóide oral (CEO) representa a neoplasia maligna mais prevalente na cavidade oral e por atingir um grande número de indivíduos, acaba se tornado um relevante problema de saúde pública. Muitos estudos demonstram alterações nos componentes da via BMP em vários tipos de tumores, como os de próstata, cólon, mama, gástricos e CEOs. É do conhecimento atual que essas proteínas podem exercer efeito pró-tumoral em estágios mais avançados do desenvolvimento neoplásico vindo a favorecer a progressão e invasão tumoral. A inibição da via de sinalização da BMP-2, através dos seus antagonistas, tem mostrado resultados positivos de ação antitumoral e que assim, o uso do Noggin pode ser um novo alvo terapêutico contra o câncer. Diante destas evidências e dos escassos trabalhos com BMP-2, Noggin e CEO, o objetivo desta pesquisa foi avaliar o efeito da BMP-2 e seu antagonista Noggin sobre a proliferação e migração celulares em culturas de células de carcinoma epidermóide de língua humana (SCC25). Foi feita a divisão em três grupos de estudo, um grupo controle, onde as células SCC25 não sofriam tratamento com substância alguma, um grupo BMP-2, no qual as células eram tratadas com 100ng/ml de BMP-2 e um grupo de células que eram tratadas com 100ng/ml de Noggin. Para o ensaio de proliferação e ciclo celular foram estabelecidos três intervalos de tempo (24, 48 e 72 horas).
A atividade proliferativa foi investigada por azul de tripan e a análise do ciclo celular através da marcação por iodeto de propídio em Citometria de fluxo. O potencial de migração/invasão das células SCC25 foi avaliado através da realização de um ensaio de invasão celular utilizando o matrigel em um intervalo de 48 horas. A curva de proliferação revelou maior crescimento celular nas células tratadas com BMP-2 no intervalo de 72 horas (p<0.05) e menor crecimento e viabilidade celular no grupo Noggin. As proteínas recombinantes favoreceram a maior porcentagem das células na fase do ciclo celular Go/G1 com diferença estatisticamente significativa no intervalo de 24 horas (p<0,05). A BMP-2 promoveu uma maior invasão das células estudadas, assim como o seu antagonista Noggin inibiu a invasão das células estudadas (p<0,05). Dessa forma, os resultados indicam que a BMP-2 favorece o fenótipo maligno, pois estimula a proliferação e invasão das células SCC25 e seu antagonista Noggin pode ser uma alternativa terapêutica pois inibiu essas características pró-tumorais. (AU)
Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin.
For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression. (AU)
Asunto(s)
Carcinoma de Células Escamosas/fisiopatología , Neoplasias de la Lengua/fisiopatología , Proliferación Celular , /efectos adversos , Citometría de Flujo/métodos , Estadísticas no Paramétricas , Metástasis de la Neoplasia , Técnicas In Vitro/métodosRESUMEN
La diferenciación mesenquimal a odontoblasto es un proceso complejo que determina la formación de los túbulos dentinales. Este proceso involucra una cuidadosa y regulada secuencia de cambios en el comportamiento de las células mesenquimales, coordinados por la expresión de diferentes factores moleculares, entre ellos, principalmente, el Noggin y BMP2. En este artículo se simula la formación de los túbulos dentinales a partir de un modelo matemático de reacción difusión que es solucionado por el método de los elementos finitos...
Mesenchymal differentiation into odontoblasts is a complex process determining the formation of dentinal tubules. The process involves a carefully regulated sequence of changes in the behavior of mesenchymal cells, coordinated by the expression of various molecular factors, particularly Noggin and BMP2. In this paper the formation of dentinal tubules is simulated using a reaction-diffusion mathematical model solved by the finite element method...
RESUMEN
Mandibular micrognathia is a deficiency in mandibular growth that prevents tooth contact during mastication, interferes with phonation and even causes sleep apnea. Studies show that mutant mice for chd (chordin) and nog (noggin) genes, which are modulators of the Bone Morphogenic Protein (BMP), had mandibular defects ranging from mandibular hypoplasia to micrognathia and agnathia. The human NOG gene was the first BMP antagonist identified and it is essential for various late events in mandibular development, which require modulation of the BMP activity. The aim of this work was to determine the presence of NOG gene polymorphisms in families with mandibular micrognathia and analyze its phenotype. Four families with mandibular micrognathia were included in this study. Blood samples were taken from the participating individuals through venipuncture and DNA was extracted. The fragments of interest were amplified using the Polymerase Chain Reaction (PCR) and the Single Nucleotide Polymorphisms (SNPs) of the NOG gene reported in the NCBI data base were analyzed through direct sequencing. The SNP rs1348322 was present in homozygote form in the subjects from all the families, where Cytosine is changed to Adenine in position 112 of the exon of the NOG gene. The SNP rs 1236187 did not show any clear result. This result suggests that there may be population polymorphism, or markers that are seldom polymorphic for our population. It is therefore necessary to continue with the search for the relationship of the NOG gene with mandibular micrognathia.
El micrognatismo mandibular, deficiencia en el crecimiento de la mandibula, no permite que los dientes entren en contacto durante la masticacion, interfiriendo con la fonacion y produciendo inclusive apnea del sueno. Estudios con ratones mutantes para el gen chordin (chd) o noggin (nog) moduladores de las proteinas morfogenicas oseas (BMP) presentaron defectos mandibulares, que van desde hipoplasia mandibular, pasando por micrognatia hasta agnatia. El gen NOG humano fue el primer antagonista de BMP identificado y es esencial para varios eventos tardios del desarrollo mandibular, que requieren modulacion de la actividad de las BMP. El objetivo del trabajo fue determinar la presencia de polimorfismos del gen NOG en pacientes con micrognatismo mandibular y analizar su fenotipo. Se tomaron 4 familias con micrognatismo mandibular, muestras de sangre fueron tomadas por venopuncion a los individuos participantes, el ADN fue extraido, se realizo la amplificacion de los fragmentos correspondientes a los polimorfismos rs 1236187 y rs 1348322 mediante PCR (Reaccion en Cadena de la Polimerasa) y se analizaron los SNPs del gen NOG reportados en la base de datos NCBI, mediante secuenciacion directa. El SNP rs 1348322, se presento en forma homocigota en los individuos de todas las familias, donde se da el cambio de una Citosina por una Adenina en la posicion 112 del exon del gen NOG. El SNP rs 1236187, no arrojo ningun resultado en forma clara. Este resultado sugiere que posiblemente pueden tratarse de polimorfismos poblacionales, o de marcadores poco polimorficos para nuestra poblacion, por lo tanto es necesario continuar en la busqueda de la relacion del gen NOG con el micrognatismo mandibular.