Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 56
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
PLoS One ; 15(1): e0227205, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31951621

RESUMO

OBJECTIVES: Low genetic diversity can lead to reduced average fitness in a population or even extinction. Preserving genetic connectivity across fragmented landscapes is therefore vital to counteract the negative consequences of genetic drift and inbreeding. This study aimed to assess the genetic composition and consequently the conservation status of a nationwide sample of European hedgehogs (Erinaceus europaeus) in Denmark. METHODS: We applied an adaptation of the genotyping by sequencing (GBS) technique to 178 individuals from six geographically distinct populations. We used a Bayesian clustering method to subdivide individuals into genetically distinct populations. We estimated individual observed (iHO), observed (HO), and unbiased expected (uHE) heterozygosity, inbreeding coefficient (FIS), percentage of polymorphic loci (P%) and tested for deviations from Hardy-Weinberg equilibrium (HWE). We used linear models to test for potential anthropogenic effects on the genetic variability of hedgehogs with iHO, uHE, P% and FIS as response variables, and assessed the demographic history of the population. RESULTS: The Danish hedgehog population is composed of three genetic clusters. We found a mean P% of 54.44-94.71, a mean uHE of 0.126-0.318 and a mean HO of 0.124-0.293 in the six populations. The FIS was found to be significantly positive for three of the six populations. We detected a large heterogeneity of iHO values within populations, which can be due to inbreeding and/or fragmentation. FIS values decreased with increasing farmland density, but there was no significant association with human population or road density. CONCLUSIONS: We found a low level of genetic variability and evidence for genetic substructure and low effective population size, which are all consequences of habitat fragmentation. We failed to detect signs of a recent population bottleneck or population increase or decline. However, because the test only identifies recent changes in population size, we cannot reject the possibility of a longer-term decline in the Danish hedgehog population.


Assuntos
Ouriços-Cacheiros/genética , Animais , Teorema de Bayes , Dinamarca , Feminino , Variação Genética , Genética Populacional , Endogamia , Masculino , Polimorfismo de Nucleotídeo Único , Densidade Demográfica
2.
Elife ; 82019 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-31036156

RESUMO

In bladder, loss of mammalian Sonic Hedgehog (Shh) accompanies progression to invasive urothelial carcinoma, but the molecular mechanisms underlying this cancer-initiating event are poorly defined. Here, we show that loss of Shh results from hypermethylation of the CpG shore of the Shh gene, and that inhibition of DNA methylation increases Shh expression to halt the initiation of murine urothelial carcinoma at the early stage of progression. In full-fledged tumors, pharmacologic augmentation of Hedgehog (Hh) pathway activity impedes tumor growth, and this cancer-restraining effect of Hh signaling is mediated by the stromal response to Shh signals, which stimulates subtype conversion of basal to luminal-like urothelial carcinoma. Our findings thus provide a basis to develop subtype-specific strategies for the management of human bladder cancer.


Assuntos
Epigênese Genética , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Ouriços-Cacheiros/genética , Transdução de Sinais/genética , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo , Idoso , Idoso de 80 Anos ou mais , Animais , Metilação de DNA , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Células Estromais/metabolismo , Células Estromais/patologia , Análise de Sobrevida , Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/patologia
3.
J Agric Food Chem ; 66(45): 11926-11934, 2018 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-30354116

RESUMO

Obesity is a risk factor for numerous metabolic disorders. In this study, we investigated the effects of the isothiocyanates sulforaphane (SA) and sulforaphene (SE) on adipogenesis in 3T3-L1 adipocytes. SE, a compound that is abundant in radish, inhibited adipogenesis by suppressing the adipogenic transcription factors peroxisome proliferator-activated receptor γ (PPARγ, 69.2 ± 2.4%, P < 0.05) and CCAAT/enhancer-binding protein α (C/EBPα, 36.1 ± 3.1%, P < 0.05), thereby reducing fat accumulation in 3T3-L1 adipocytes (45.6 ± 2.7%, P < 0.05); SA was less effective. SE exerted these activities through the activation of the Hedgehog (Hh) signaling pathway by restoring Smo ((2.1 ± 0.2)-fold, P < 0.05) and Gli1 ((2.8 ± 0.1)-fold, P < 0.05) expression, which was suppressed by adipogenic signals. These effects of SE were abrogated by treatment with the Hh inhibitor vismodegib. Thus, SE inhibits adipocyte differentiation via Hh signaling and may be an effective natural agent for preventing adipocyte hyperplasia and obesity.


Assuntos
Adipócitos/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Fármacos Antiobesidade/farmacologia , Ouriços-Cacheiros/metabolismo , Isotiocianatos/farmacologia , Células 3T3-L1 , Adipócitos/citologia , Adipócitos/metabolismo , Animais , Proteína alfa Estimuladora de Ligação a CCAAT/genética , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Ouriços-Cacheiros/genética , Camundongos , PPAR gama/genética , PPAR gama/metabolismo , Transdução de Sinais/efeitos dos fármacos
4.
J Cell Sci ; 131(15)2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29930086

RESUMO

Hedgehog (Hh) transduces signals by promoting cell surface accumulation and activation of the G-protein-coupled receptor (GPCR)-family protein Smoothened (Smo) in Drosophila, but the molecular mechanism underlying the regulation of Smo trafficking remains poorly understood. Here, we identified the Cul4-DDB1 E3 ubiquitin ligase complex as being essential for Smo ubiquitylation and cell surface clearance. We found that the C-terminal intracellular domain of Smo recruits Cul4-DDB1 through the ß subunit of trimeric G protein (Gß), and that Cul4-DDB1-Gß promotes the ubiquitylation of both Smo and its binding partner G-protein-coupled-receptor kinase 2 (Gprk2) and induces the internalization and degradation of Smo. Hh dissociates Cul4-DDB1 from Smo by recruiting the catalytic subunit of protein kinase A (PKA) to phosphorylate DDB1, which disrupts its interaction with Gß. Inactivation of the Cul4-DDB1 complex resulted in elevated Smo cell surface expression, whereas an excessive amount of Cul4-DDB1 blocked Smo accumulation and attenuated Hh pathway activation. Taken together, our study identifies an E3 ubiquitin ligase complex targeting Smo for ubiquitylation and provides new insight into how Hh signaling regulates Smo trafficking and cell surface expression.


Assuntos
Proteínas Culina/metabolismo , Proteínas de Drosophila/metabolismo , Transdução de Sinais/fisiologia , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação/fisiologia , Animais , Proteínas Culina/genética , Proteínas de Drosophila/genética , Ouriços-Cacheiros/genética , Ouriços-Cacheiros/metabolismo , Ligação Proteica , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Ubiquitina/genética , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/genética
5.
Zool Res ; 39(5): 335-347, 2018 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-29695683

RESUMO

Hedgehogs in the genus Mesechinus (Family Erinaceidae), which include two currently recognized species (M. dauuricus and M. hughi), are distributed from northeast Mongolia to the upper Amur Basin in Russia and adjacent areas in northeast and northern China. In recent years, a population of Mesechinus hedgehogs was discovered from Mt. Gaoligong, southwestern Yunnan, China, far from the known distribution range of the genus. Furthermore, these hedgehogs are the only known population to be distributed at elevations higher than 2 100 m and in sympatry with gymnures. To evaluate the taxonomic status of these hedgehogs, we examined specimens representing Mesechinus taxa in China and further conducted morphometric and karyotypic analyses. Our results supported the existence of four species in China. Specifically, we identified the hedgehogs from Mt. Gaoligong as a new species, Mesechinus wangi sp. nov., and recognized M. miodon, previously considered as a synonym of either M. dauuricus or M. hughi, as a distinct species. Interestingly, we observed a supernumerary M4 on all specimens of Mesechinus wangi sp. nov., which is an extremely rare event in the evolution of mammalian dentition.


Assuntos
Ouriços-Cacheiros/classificação , Animais , China , Demografia , Ecossistema , Ouriços-Cacheiros/anatomia & histologia , Ouriços-Cacheiros/genética , Cariotipagem , Mongólia , Sibéria
6.
Exp Neurol ; 303: 72-79, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29337143

RESUMO

BACKGROUND: Facial nerve paralysis is a significant cause of morbidity, affecting facial appearance, emotional expression, speech, oral competence, and vision. A more complete understanding of the complex cellular events required for successful nerve regeneration may reveal new therapeutic targets. The role of fibroblasts in regeneration, and the process by which the nerve reforms its three-dimensional structure after a transection injury, are not fully understood. The Hedgehog signaling pathway has been shown to mediate nerve sheath formation during development. We therefore sought to characterize the role of Hedgehog-responsive cells following transection of the facial nerve. METHODS: Two transgenic mouse lines with reporters for the downstream effector of Hedgehog signaling, Gli1, were used. The animals underwent a unilateral facial nerve transection injury, and the contralateral side served as a control. Facial nerves were analyzed via immunohistochemistry and immunofluorescence at predetermined time points as the facial nerve regenerated after the transection injury. RESULTS: There was a statistically significant increase in Gli1+ cells both at the site of injury and within the distal nerve segment over time. Gli1+ cells are fibroblasts within the nerve and appear to contribute to the reformation of the nerve sheath after injury. CONCLUSION: These findings describe a key signaling pathway by which fibroblasts participate in motor nerve regeneration. Fibroblasts that reside within the nerve respond to injury and may represent a novel therapeutic target in the context of facial nerve regeneration after transection injury.


Assuntos
Traumatismos do Nervo Facial/patologia , Fibroblastos/metabolismo , Ouriços-Cacheiros/metabolismo , Regeneração Nervosa/genética , Proteína GLI1 em Dedos de Zinco/metabolismo , Animais , Antígenos/metabolismo , Modelos Animais de Doenças , Fibronectinas/metabolismo , Citometria de Fluxo , Galactosídeos/genética , Galactosídeos/metabolismo , Regulação da Expressão Gênica/genética , Ouriços-Cacheiros/genética , Indóis/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/metabolismo , Neurônios/patologia , Neurônios/ultraestrutura , Proteoglicanas/metabolismo , Receptor de Fator de Crescimento Neural/metabolismo , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo , Transdução de Sinais/genética , Proteína GLI1 em Dedos de Zinco/genética
7.
JCI Insight ; 2(21)2017 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-29093271

RESUMO

Advanced basal cell carcinomas (BCCs) circumvent Smoothened (SMO) inhibition by activating GLI transcription factors to sustain the high levels of Hedgehog (HH) signaling required for their survival. Unfortunately, there is a lack of efficacious therapies. We performed a gene expression-based drug repositioning screen in silico and identified the FDA-approved histone deacetylase (HDAC) inhibitor, vorinostat, as a top therapeutic candidate. We show that vorinostat only inhibits proliferation of BCC cells in vitro and BCC allografts in vivo at high dose, limiting its usefulness as a monotherapy. We leveraged this in silico approach to identify drug combinations that increase the therapeutic window of vorinostat and identified atypical PKC Ɩ/ʎ (aPKC) as a HDAC costimulator of HH signaling. We found that aPKC promotes GLI1-HDAC1 association in vitro, linking two positive feedback loops. Combination targeting of HDAC1 and aPKC robustly inhibited GLI1, lowering drug doses needed in vitro, in vivo, and ex vivo in patient-derived BCC explants. We identified a bioavailable and selective small-molecule aPKC inhibitor, bringing the pharmacological blockade of aPKC and HDAC1 into the realm of clinical possibility. Our findings provide a compelling rationale and candidate drugs for combined targeting of HDAC1 and aPKC in HH-dependent cancers.


Assuntos
Carcinoma Basocelular/tratamento farmacológico , Histona Desacetilase 1/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Isoenzimas/efeitos dos fármacos , Proteína Quinase C/efeitos dos fármacos , Neoplasias Cutâneas/tratamento farmacológico , Aloenxertos , Animais , Carcinoma Basocelular/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Biologia Computacional , Combinação de Medicamentos , Descoberta de Drogas , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Ouriços-Cacheiros/genética , Ouriços-Cacheiros/metabolismo , Histona Desacetilase 1/genética , Histona Desacetilase 1/metabolismo , Inibidores de Histona Desacetilases/química , Isoenzimas/metabolismo , Camundongos , Camundongos Knockout , Proteína Quinase C/metabolismo , Transdução de Sinais , Fatores de Transcrição/efeitos dos fármacos , Fatores de Transcrição/genética , Proteína GLI1 em Dedos de Zinco/genética , Proteína GLI1 em Dedos de Zinco/metabolismo
8.
Gene ; 620: 54-65, 2017 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-28400268

RESUMO

Traditional subspecies call attention to differences between geographic populations with research potential, but their value is often in need of revision. Genetic data can be useful for evaluating the taxonomic validity of historical species and subspecies designations or for identifying morphologically cryptic divergent lineages worthy of further in-depth taxonomic study. The desert hedgehog (Paraechinus aethiopicus) has a wide but fragmented distribution in arid and semi-arid habitats from the northwest to the northeast of Africa and southwestern Asia, and its taxonomy is still unclear. We used mitochondrial (cytochrome b, Cyt b, and 12S ribosomal RNA, 12S) and nuclear (breast cancer type 1 susceptibility protein, BRCA1, and apolipoprotein B, Apob) DNA sequence data to assess the degree of genetic divergence between two of its three major proposed subspecies: Arabian (P. a. dorsalis) and Northwest African (P. a. deserti); this is the first molecular evaluation of the taxonomy of P. aethiopicus. Phylogenetic analyses, comparison of interspecific and intraspecific genetic distances observed across hedgehog species, and molecular species delimitation methods (distance-based clustering and tree-based), all indicate a level of genetic differentiation between dorsalis and deserti that is compatible with their taxonomic separation. Their divergence in the studied genes were consistently comparable to, or greater than, several intrageneric and a few intergeneric distances in hedgehogs. The Cyt b net Kimura 2-parameter distance between dorsalis and deserti was 10.8±1.3%, which is about the mean between congeneric species in reviews of Cyt b distances for mammals. This study, as a test of the genetic distinctiveness of dorsalis and deserti, suggests that they represent evolutionarily significant units and flags them for future phylogeographic and taxonomic investigations.


Assuntos
Especiação Genética , Ouriços-Cacheiros/genética , África , Animais , Apolipoproteínas B/genética , Proteína BRCA1/genética , Citocromos b/genética , Evolução Molecular , Ouriços-Cacheiros/classificação , Filogenia , RNA Ribossômico/genética
9.
Genet Mol Res ; 16(1)2017 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-28198504

RESUMO

We sequenced and characterized the complete mitogenome (KX964606) of the Amur hedgehog Erinaceus amurensis to provide more data for comparative mitogenomics of the genus Erinaceus (Erinaceidae). The mitogenome of E. amurensis is a circular molecule 16,941 bp long, consisting of a control region and a conserved set of 37 genes containing 13 protein-coding genes, 22 tRNA genes, and two rRNA genes (12S rRNA and 16S rRNA). The mitogenome of E. amurensis is AT-biased, with a nucleotide composition of 33.9% A, 21.1% C, 32.6% T, and 12.4% G. The mitogenomes of E. amurensis and the closely related hedgehog species E. europaeus, excluding the control region (66.7%), share over 90% sequence similarity. According to the inter-generic relationship based on six mitogenomes described from five genera of Erinaceidae, the subfamilies Erinaceinae and Galericinae are strongly supported as monophyletic groups, with each genus well placed within its own subfamily. Within the subfamily Erinaceinae, E. amurensis is a sister species to E. europaeus, and the relationship between Hemiechinus and Erinaceus is strongly supported. Within the subfamily Galericinae, the clade of Hylomys + Neotetracus was sister to that of Echinosorex, with clades supported by high values. Our findings will help to understand the codon usage pattern and molecular evolution of E. amurensis, and provide insight into inter-generic relationships within the family Erinaceidae. In future studies, the inclusion of mitogenomes from other genera would greatly enhance our understanding of higher phylogeny within the Erinaceidae.


Assuntos
Genoma Mitocondrial , Ouriços-Cacheiros/classificação , Ouriços-Cacheiros/genética , Filogenia , Animais , Fases de Leitura Aberta , RNA Ribossômico/genética , RNA Ribossômico 16S/genética , RNA de Transferência/genética
10.
PLoS Negl Trop Dis ; 11(2): e0005137, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28231240

RESUMO

Schistosomiasis affects approximately 240 million people in the world. Schistosoma mansoni eggs in the liver induce periportal fibrosis and hepatic failure driven by monocyte recruitment and macrophage activation, resulting in robust Th2 response. Here, we suggested a possible involvement of Galectin-3 (Gal-3), histone deacetylases (HDACs), and Hedgehog (Hh) signaling with macrophage activation during Th1/Th2 immune responses, fibrogranuloma reaction, and tissue repair during schistosomiasis. Gal-3 is highly expressed by liver macrophages (Kupffer cells) around Schistosoma eggs. HDACs and Hh regulate macrophage polarization and hepatic stellate cell activation during schistosomiasis-associated fibrogenesis. Previously, we demonstrated an abnormal extracellular matrix distribution in the liver that correlated with atypical monocyte-macrophage differentiation in S. mansoni-infected, Gal-3-deficient (Lgals3-/-) mice. New findings explored in this review focus on the chronic phase, when wild-type (Lgals3+/+) and Lgals3-/- mice were analyzed 90 days after cercariae infection. In Lgals3-/- infected mice, there was significant inflammatory infiltration with myeloid cells associated with egg destruction (hematoxylin and eosin staining), phagocytes (specifically Kupffer cells), numerically reduced and diffuse matrix extracellular deposition in fibrotic areas (Gomori trichrome staining), and severe disorganization of collagen fibers surrounding the S. mansoni eggs (reticulin staining). Granuloma-derived stromal cells (GR cells) of Lgals3-/- infected mice expressed lower levels of alpha smooth muscle actin (α-SMA) and eotaxin and higher levels of IL-4 than Lgals3+/+ mice (real-time PCR). The relevant participation of macrophages in these events led us to suggest distinct mechanisms of activation that culminate in defective fibrosis in the liver of Lgals3-/- infected mice. These aspects were discussed in this review, as well as the possible interference between Gal-3, HDACs, and Hh signaling during progressive liver fibrosis in S. mansoni-infected mice. Further studies focused on macrophage roles could elucidate these questions and clear the potential utility of these molecules as antifibrotic targets.


Assuntos
Galectina 3/metabolismo , Ouriços-Cacheiros/metabolismo , Histona Desacetilases/metabolismo , Cirrose Hepática/metabolismo , Esquistossomose/complicações , Animais , Galectina 3/genética , Ouriços-Cacheiros/genética , Histona Desacetilases/genética , Humanos , Cirrose Hepática/etiologia , Cirrose Hepática/genética , Esquistossomose/parasitologia , Esquistossomose Japônica/parasitologia , Transdução de Sinais
11.
Nat Prod Commun ; 11(6): 747-8, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27534107

RESUMO

Two new 13, 14/14, 15-disecopregnane-type skeleton C21 steroidal aglycones, neocynapanogenin G (1) and neocynapanogenin H (2), were isolated from the hydrolyzed extract of the CHCl3 soluble extract of the roots of Cynanchun paniculatum. Their structures were determined on the basis of chemical evidence and extensive spectroscopic methods, including 1D and 2D NMR spectroscopy. Compound 1 displayed signifidant inhibition of the Hedgehog signaling pathway in vitro.


Assuntos
Cynanchum/química , Medicamentos de Ervas Chinesas/química , Iridoides/química , Raízes de Plantas/química , Esteroides/química , Animais , Linhagem Celular , Ouriços-Cacheiros/genética , Ouriços-Cacheiros/metabolismo , Humanos , Estrutura Molecular , Transdução de Sinais/efeitos dos fármacos
12.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(6): 4712-4714, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-26678715

RESUMO

The complete mitochondrial genome of the small-toothed forest hedgehog Mesechinus miodon (Eulipotyphla: Erinaceidae) has been reconstructed from Illumina sequencing data. The circular genome is 16 842 bp long, comprising 22 transfer RNAs (tRNAs), 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), and one control region. All PCGs are initiated with ATR (ATA/ATG) codons, except for nad4 with GTG as its initiation codon. Two PCGs (cox3 and nad4) harbor an incomplete termination codon T, while the others are terminated with TAA (atp6, cox1, cox2, nad1, nad2, nad3, nad4l, nad5, and nad6), TAG (atp8) or AGA (cytb). The base composition is highly biased (34.9% A, 19.7% C, 11.9% G, and 33.5% T for the light strand) with an overall A + T content of 68.4%. Phylogenetic analysis indicated that M. miodon is more closely related to the consubfamilial long-eared hedgehog Hemiechinus auritus than to those within the order Eulipotyphla.


Assuntos
Genes Mitocondriais , Genoma Mitocondrial , Ouriços-Cacheiros/genética , Animais , Composição de Bases , Códon de Iniciação , Códon de Terminação , Evolução Molecular , Ouriços-Cacheiros/classificação , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Análise de Sequência de DNA
13.
Artigo em Inglês | MEDLINE | ID: mdl-26393434

RESUMO

Bats exhibit higher paracellular absorption of glucose-sized molecules than non-flying mammals, a phenomenon that may be driven by higher permeability of the intestinal tight junctions. The various claudins, occludin, and other proteins making up the tight junctions are thought to determine their permeability properties. Here we show that absorption of the paracellular probe l-arabinose is higher in a bat (Eptesicus fuscus) than in a vole (Microtus pennsylvanicus) or a hedgehog (Atelerix albiventris). Furthermore, histological measurements demonstrated that hedgehogs have many more enterocytes in their intestines, suggesting that bats cannot have higher absorption of arabinose simply by having more tight junctions. We therefore investigated the mRNA levels of several claudins and occludin, because these proteins may affect permeability of tight junctions to macronutrients. To assess the expression levels of claudins per tight junction, we normalized the mRNA levels of the claudins to the constitutively expressed tight junction protein ZO-1, and combined these with measurements previously made in a bat and a rodent to determine if there were among-species differences. Although expression ratios of several genes varied among species, there was not a consistent difference between bats and non-flyers in the expression ratio of any particular gene. Protein expression patterns may differ from mRNA expression patterns, and might better explain differences among species in arabinose absorption.


Assuntos
Claudinas/genética , Regulação da Expressão Gênica , Absorção Intestinal , Mucosa Intestinal/metabolismo , Intestinos/citologia , Animais , Arvicolinae/genética , Arvicolinae/metabolismo , Quirópteros/genética , Quirópteros/metabolismo , Ouriços-Cacheiros/genética , Ouriços-Cacheiros/metabolismo , Especificidade da Espécie
14.
PLoS One ; 10(5): e0126562, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25978455

RESUMO

Comprehensive analysis of alterations in gene expression along with neoplastic transformation in human cells provides valuable information about the molecular mechanisms underlying transformation. To further address these questions, we performed whole transcriptome analysis to the human mesenchymal stem cell line, UE6E7T-3, which was immortalized with hTERT and human papillomavirus type 16 E6/E7 genes, in association with progress of transformation in these cells. At early stages of culture, UE6E7T-3 cells preferentially lost one copy of chromosome 13, as previously described; in addition, tumor suppressor genes, DNA repair genes, and apoptosis-activating genes were overexpressed. After the loss of chromosome 13, additional aneuploidy and genetic alterations that drove progressive transformation, were observed. At this stage, the cell line expressed oncogenes as well as genes related to anti-apoptotic functions, cell-cycle progression, and chromosome instability (CIN); these pro-tumorigenic changes were concomitant with a decrease in tumor suppressor gene expression. At later stages after prolong culture, the cells exhibited chromosome translocations, acquired anchorage-independent growth and tumorigenicity in nude mice, (sarcoma) and exhibited increased expression of genes encoding growth factor and DNA repair genes, and decreased expression of adhesion genes. In particular, glypican-5 (GPC5), which encodes a cell-surface proteoglycan that might be a biomarker for sarcoma, was expressed at high levels in association with transformation. Patched (Ptc1), the cell surface receptor for hedgehog (Hh) signaling, was also significantly overexpressed and co-localized with GPC5. Knockdown of GPC5 expression decreased cell proliferation, suggesting that it plays a key role in growth in U3-DT cells (transformants derived from UE6E7T-3 cells) through the Hh signaling pathway. Thus, the UE6E7T-3 cell culture model is a useful tool for assessing the functional contribution of genes showed by expression profiling to the neoplastic transformation of human fibroblasts and human mesenchymal stem cells (hMSC).


Assuntos
Transformação Celular Neoplásica/genética , Transformação Celular Viral/genética , Células-Tronco Mesenquimais/metabolismo , Transcrição Genética/genética , Aneuploidia , Animais , Técnicas de Cultura de Células , Ciclo Celular/genética , Proliferação de Células/genética , Instabilidade Cromossômica/genética , Cromossomos Humanos Par 13/genética , Reparo do DNA/genética , Fibroblastos/metabolismo , Glipicanas/genética , Ouriços-Cacheiros/genética , Papillomavirus Humano 6/genética , Humanos , Camundongos , Camundongos Nus , Oncogenes/genética , Transdução de Sinais/genética , Telomerase/genética , Ativação Transcricional/imunologia
15.
J Agric Food Chem ; 62(17): 3759-67, 2014 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-24724627

RESUMO

In breast cancer, the cytokine tumor necrosis factor-α (TNF-α) induces cell invasion, although the molecular basis of it has not been clearly elucidated. In this study, we investigated the role of daidzein in regulating TNF-α induced cell invasion and the underlying molecular mechanisms. Daidzein inhibited TNF-α induced cellular migration and invasion in estrogen receptor (ER) negative MCF10DCIS.com human breast cancer cells. TNF-α activated Hedgehog (Hh) signaling by enhancing Gli1 nuclear translocation and transcriptional activity, which resulted in increased invasiveness; these effects were blocked by daidzein and the Hh signaling inhibitors, cyclopamine and vismodegib. Moreover, these compounds suppressed TNF-α induced matrix metalloproteinase (MMP)-9 mRNA expression and activity. Taken together, mammary tumor cell invasiveness was stimulated by TNF-α induced activation of Hh signaling; these effects were abrogated by daidzein, which suppressed Gli1 activation, thereby inhibiting migration and invasion.


Assuntos
Neoplasias da Mama/fisiopatologia , Ouriços-Cacheiros/metabolismo , Isoflavonas/farmacologia , Proteínas Oncogênicas/metabolismo , Extratos Vegetais/farmacologia , Transdução de Sinais/efeitos dos fármacos , Soja/química , Transativadores/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Nucléolo Celular/genética , Nucléolo Celular/metabolismo , Regulação para Baixo/efeitos dos fármacos , Feminino , Ouriços-Cacheiros/genética , Humanos , Invasividade Neoplásica/genética , Proteínas Oncogênicas/genética , Transativadores/genética , Fator de Necrose Tumoral alfa/genética , Proteína GLI1 em Dedos de Zinco
16.
Mol Ecol ; 22(14): 3709-20, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23711046

RESUMO

The crucial steps in biological invasions, related to the shaping of genetic architecture and the current evolution of adaptations to a novel environment, usually occur in small populations during the phases of introduction and establishment. However, these processes are difficult to track in nature due to invasion lag, large geographic and temporal scales compared with human observation capabilities, the frequent depletion of genetic variance, admixture and other phenomena. In this study, we compared genetic and historical evidence related to the invasion of the West European hedgehog to New Zealand to infer details about the introduction and establishment. Historical information indicates that the species was initially established on the South Island. A molecular assay of populations from Great Britain and New Zealand using mitochondrial sequences and nuclear microsatellite loci was performed based on a set of analyses including approximate Bayesian computation, a powerful approach for disentangling complex population demographies. According to these analyses, the population of the North Island was most similar to that of the native area and showed greatest reduction in genetic variation caused by founder demography and/or drift. This evidence indicated the location of the establishment phase. The hypothesis was corroborated by data on climate and urbanization. We discuss the contrasting results obtained by the molecular and historical approaches in the light of their different explanatory power and the possible biases influencing the description of particular aspects of invasions, and we advocate the integration of the two types of approaches in invasion biology.


Assuntos
DNA Mitocondrial/genética , Genética Populacional , Ouriços-Cacheiros/fisiologia , Animais , Teorema de Bayes , Ouriços-Cacheiros/genética , Humanos , Espécies Introduzidas , Repetições de Microssatélites/genética , Dados de Sequência Molecular , Nova Zelândia , Densidade Demográfica
17.
Prostate Cancer Prostatic Dis ; 16(3): 233-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23670255

RESUMO

BACKGROUND: Loss or mutations of the BRCA1 gene are associated with increased risk of breast and ovarian cancers and with prostate cancer (PCa) aggressiveness. Previously, we identified GADD153 as a target of BRCA1 protein, which increases doxorubicin sensitivity in human p53 -/- PCa cells (PC3). Considering that p53 is a crucial target in cancer therapy, in this work we investigated p53 role in the regulation of transcription of GADD153. METHODS: We performed reverse transcription quantitative PCR (RT-qPCR), western blot and luciferase assays to analyze GADD153 and/or BRCA1 expression in response to ultraviolet or doxorubicin exposure in PC3 p53 stable-transfected cells and LNCaP (p53+/+) cells. BRCA1 protein recruitment to GADD153 promoter was studied by chromatin immunoprecipitation-qPCR. To assess expression of BRCA1 and/or p53 target genes, we used a panel of stable-transfected PCa cell lines. We finally analyzed these genes in vivo using BRCA1-depleted PCa xenograft models. RESULTS: We found that GADD153 was highly induced by doxorubicin in PC3 cells; however, this response was totally abolished in LNCaP (p53wt) and in p53-restituted PC3 cells. Furthermore, BRCA1 protein associates to GADD153 promoter after DNA damage in the presence of p53. Additionally, we demonstrated that BRCA1 and/or p53 modulate genes involved in DNA damage and cell cycle regulation (cyclin D1, BLM, BRCA2, DDB2, p21(WAF1/CIP1), H3F3B, GADD153, GADD45A, FEN1, CCNB2), EMT (E-cadherin, ß-catenin, vimentin, fibronectin, slug, snail) and Hedgehog pathways (SHH, IHH, DHH, Gli1, PATCH1). Furthermore, xenograft studies demonstrated that BRCA1 knockdown in PC3 cells increased tumor growth and modulated these genes in vivo. CONCLUSIONS: Although BRCA1 induces GADD153 in a p53 independent manner, p53 abolished GADD153 induction in response to DNA damage. In addition, several important PCa targets are modulated by BRCA1 and p53. Altogether, these data might be important to understand the therapy response of PCa patients.


Assuntos
Proteína BRCA1/metabolismo , Neoplasias da Próstata/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Proteína BRCA1/genética , Ciclo Celular/genética , Linhagem Celular Tumoral , Dano ao DNA , Ouriços-Cacheiros/genética , Ouriços-Cacheiros/metabolismo , Xenoenxertos , Humanos , Masculino , Camundongos , Neoplasias da Próstata/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fator de Transcrição CHOP/metabolismo , Transcrição Genética , Proteína Supressora de Tumor p53/genética
18.
PLoS One ; 7(6): e39304, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22745729

RESUMO

BACKGROUND: Erinaceidae is a family of small mammals that include the spiny hedgehogs (Erinaceinae) and the silky-furred moonrats and gymnures (Galericinae). These animals are widely distributed across Eurasia and Africa, from the tundra to the tropics and the deserts to damp forests. The importance of these animals lies in the fact that they are the oldest known living placental mammals, which are well represented in the fossil record, a rarity fact given their size and vulnerability to destruction during fossilization. Although the Family has been well studied, their phylogenetic relationships remain controversial. To test previous phylogenetic hypotheses, we combined molecular and morphological data sets, including representatives of all the genera. METHODOLOGY AND PRINCIPAL FINDINGS: We included in the analyses 3,218 bp mitochondrial genes, one hundred and thirty-five morphological characters, twenty-two extant erinaceid taxa, and five outgroup taxa. Phylogenetic relationships were reconstructed using both partitioned and combined data sets. As in previous analyses, our results strongly support the monophyly of both subfamilies (Galericinae and Erinaceinae), the Hylomys group (to include Neotetracus and Neohylomys), and a sister-relationship of Atelerix and Erinaceus. As well, we verified that the extremely long branch lengths within the Galericinae are consistent with their fossil records. Not surprisingly, we found significant incongruence between the phylogenetic signals of the genes and the morphological characters, specifically in the case of Hylomys parvus, Mesechinus, and relationships between Hemiechinus and Paraechinus. CONCLUSIONS: Although we discovered new clues to understanding the evolutionary relationships within the Erinaceidae, our results nonetheless, strongly suggest that more robust analyses employing more complete taxon sampling (to include fossils) and multiple unlinked genes would greatly enhance our understanding of the Erinaceidae. Until then, we have left the nomenclature of the taxa unchanged; hence it does not yet precisely reflect their phylogenetic relationships or the depth of their genetic diversity.


Assuntos
Ouriços-Cacheiros/genética , Filogenia , Animais , Variação Genética/genética , Ouriços-Cacheiros/classificação
19.
Mol Biol Rep ; 39(5): 6123-32, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22246941

RESUMO

Here we describe the identification of the hedgehog Erinaceus europaeus homologue of a proliferation-inducing ligand (APRIL) of the TNF family (designated heAPRIL). Hedgehog APRIL contains two cysteine residues (Cys(196) and Cys(211)), a furin protease cleavage site and a conserved putative N-glycosylation site (Asn(124)). Real-time quantitative PCR (qPCR) analysis revealed that heAPRIL could be detected in various tissues. MTT assays and flow cytometric analysis revealed that Nus-hesAPRIL and hesAPRIL could promote the survival/proliferation of splenic B cells. Laser scanning confocal microscopy analysis showed GFP-hesAPRIL could successfully bind to the APRIL receptors of lymphocytes.


Assuntos
Regulação da Expressão Gênica , Ouriços-Cacheiros/genética , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/genética , Aminoácidos/metabolismo , Animais , Linfócitos B/citologia , Proliferação de Células , Sobrevivência Celular , Clonagem Molecular , Perfilação da Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos ICR , Microscopia Confocal , Microscopia de Fluorescência , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Homologia de Sequência de Aminoácidos , Baço/citologia , Homologia Estrutural de Proteína , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/química , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo
20.
Heredity (Edinb) ; 108(3): 248-55, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21863052

RESUMO

We used the mitochondrial control region and nuclear microsatellites to assess the distribution patterns, population structure, demography and landscape genetics for the hedgehogs Erinaceus europaeus and Erinaceus roumanicus in a transect of the mid-European zone of sympatry. E. roumanicus was less frequent and restricted to regions with lower altitudes. Demographic analyses suggested recent population growth in this species. A comparison of patterns in the spatial variability of mitochondrial and nuclear DNA indicated less sex-biased dispersal and higher levels of gene flow in E. roumanicus. No evidence of recent hybridisation or introgression was detected. We interpreted these results by comparing with phylogeographic and palaeontological studies as well as with the occurrence of hybridisation in the Russian contact zone. We propose that Central Europe was colonised by E. roumanicus by the beginning of the Neolithic period and that there was a subsequent reinforcement stage as well as the formation of a zone of sympatry after the complete reproductive isolation of both species.


Assuntos
Genética Populacional , Ouriços-Cacheiros/genética , Simpatria , Animais , DNA Mitocondrial , Demografia , Europa (Continente) , Feminino , Fluxo Gênico , Variação Genética , Haplótipos , Masculino , Repetições de Microssatélites , Filogeografia , Isolamento Reprodutivo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA