Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 54
Filtrar
Mais filtros

Base de dados
Tipo de documento
Intervalo de ano de publicação
1.
Mol Cell ; 84(19): 3826-3842.e8, 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39303722

RESUMO

RNA interactome studies have revealed that hundreds of zinc-finger proteins (ZFPs) are candidate RNA-binding proteins (RBPs), yet their RNA substrates and functional significance remain largely uncharacterized. Here, we present a systematic multi-omics analysis of the DNA- and RNA-binding targets and regulatory roles of more than 100 ZFPs representing 37 zinc-finger families. We show that multiple ZFPs are previously unknown regulators of RNA splicing, alternative polyadenylation, stability, or translation. The examined ZFPs show widespread sequence-specific RNA binding and preferentially bind proximal to transcription start sites. Additionally, several ZFPs associate with their targets at both the DNA and RNA levels. We highlight ZNF277, a C2H2 ZFP that binds thousands of RNA targets and acts as a multi-functional RBP. We also show that ZNF473 is a DNA/RNA-associated protein that regulates the expression and splicing of cell cycle genes. Our results reveal diverse roles for ZFPs in transcriptional and post-transcriptional gene regulation.


Assuntos
Splicing de RNA , Proteínas de Ligação a RNA , Dedos de Zinco , Humanos , Proteínas de Ligação a RNA/metabolismo , Proteínas de Ligação a RNA/genética , Regulação da Expressão Gênica , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/genética , RNA/metabolismo , RNA/genética , Ligação Proteica , Células HEK293 , Células HeLa , Processamento Alternativo , Sítios de Ligação , Multiômica
2.
Nature ; 583(7818): 711-719, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32728246

RESUMO

Many proteins regulate the expression of genes by binding to specific regions encoded in the genome1. Here we introduce a new data set of RNA elements in the human genome that are recognized by RNA-binding proteins (RBPs), generated as part of the Encyclopedia of DNA Elements (ENCODE) project phase III. This class of regulatory elements functions only when transcribed into RNA, as they serve as the binding sites for RBPs that control post-transcriptional processes such as splicing, cleavage and polyadenylation, and the editing, localization, stability and translation of mRNAs. We describe the mapping and characterization of RNA elements recognized by a large collection of human RBPs in K562 and HepG2 cells. Integrative analyses using five assays identify RBP binding sites on RNA and chromatin in vivo, the in vitro binding preferences of RBPs, the function of RBP binding sites and the subcellular localization of RBPs, producing 1,223 replicated data sets for 356 RBPs. We describe the spectrum of RBP binding throughout the transcriptome and the connections between these interactions and various aspects of RNA biology, including RNA stability, splicing regulation and RNA localization. These data expand the catalogue of functional elements encoded in the human genome by the addition of a large set of elements that function at the RNA level by interacting with RBPs.


Assuntos
Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/metabolismo , Transcriptoma/genética , Processamento Alternativo/genética , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Cromatina/genética , Cromatina/metabolismo , Bases de Dados Genéticas , Feminino , Técnicas de Silenciamento de Genes , Humanos , Espaço Intracelular/genética , Masculino , Ligação Proteica , RNA Mensageiro/química , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/genética , Especificidade por Substrato
3.
Nucleic Acids Res ; 51(D1): D1549-D1557, 2023 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-36321651

RESUMO

RNA binding proteins (RBPs) are central regulators of gene expression implicated in all facets of RNA metabolism. As such, they play key roles in cellular physiology and disease etiology. Since different steps of post-transcriptional gene expression tend to occur in specific regions of the cell, including nuclear or cytoplasmic locations, defining the subcellular distribution properties of RBPs is an important step in assessing their potential functions. Here, we present the RBP Image Database, a resource that details the subcellular localization features of 301 RBPs in the human HepG2 and HeLa cell lines, based on the results of systematic immuno-fluorescence studies conducted using a highly validated collection of RBP antibodies and a panel of 12 markers for specific organelles and subcellular structures. The unique features of the RBP Image Database include: (i) hosting of comprehensive representative images for each RBP-marker pair, with ∼250,000 microscopy images; (ii) a manually curated controlled vocabulary of annotation terms detailing the localization features of each factor; and (iii) a user-friendly interface allowing the rapid querying of the data by target or annotation. The RBP Image Database is freely available at https://rnabiology.ircm.qc.ca/RBPImage/.


Assuntos
Bases de Dados Factuais , Imagem Óptica , Proteínas de Ligação a RNA , Humanos , Anticorpos/metabolismo , Células HeLa , RNA/química , Proteínas de Ligação a RNA/metabolismo , Células Hep G2
4.
Mol Cell ; 61(6): 821-33, 2016 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-26907613

RESUMO

Spatial restriction of mRNA to distinct subcellular locations enables local regulation and synthesis of proteins. However, the organizing principles of mRNA localization remain poorly understood. Here we analyzed subcellular transcriptomes of neural projections and soma of primary mouse cortical neurons and two neuronal cell lines and found that alternative last exons (ALEs) often confer isoform-specific localization. Surprisingly, gene-distal ALE isoforms were four times more often localized to neurites than gene-proximal isoforms. Localized isoforms were induced during neuronal differentiation and enriched for motifs associated with muscleblind-like (Mbnl) family RNA-binding proteins. Depletion of Mbnl1 and/or Mbnl2 reduced localization of hundreds of transcripts, implicating Mbnls in localization of mRNAs to neurites. We provide evidence supporting a model in which the linkage between genomic position of ALEs and subcellular localization enables coordinated induction of localization-competent mRNA isoforms through a post-transcriptional regulatory program that is induced during differentiation and reversed in cellular reprogramming and cancer.


Assuntos
Proteínas de Ligação a DNA/genética , Neuritos/metabolismo , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genética , Processamento Alternativo/genética , Animais , Diferenciação Celular/genética , Reprogramação Celular/genética , Proteínas de Ligação a DNA/antagonistas & inibidores , Éxons , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Camundongos , Isoformas de Proteínas , Estrutura Terciária de Proteína , Processamento Pós-Transcricional do RNA/genética , Proteínas de Ligação a RNA/antagonistas & inibidores , Transcriptoma/genética
5.
Mol Cell ; 61(6): 903-13, 2016 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-26990993

RESUMO

Transcriptome-wide maps of RNA binding protein (RBP)-RNA interactions by immunoprecipitation (IP)-based methods such as RNA IP (RIP) and crosslinking and IP (CLIP) are key starting points for evaluating the molecular roles of the thousands of human RBPs. A significant bottleneck to the application of these methods in diverse cell lines, tissues, and developmental stages is the availability of validated IP-quality antibodies. Using IP followed by immunoblot assays, we have developed a validated repository of 438 commercially available antibodies that interrogate 365 unique RBPs. In parallel, 362 short-hairpin RNA (shRNA) constructs against 276 unique RBPs were also used to confirm specificity of these antibodies. These antibodies can characterize subcellular RBP localization. With the burgeoning interest in the roles of RBPs in cancer, neurobiology, and development, these resources are invaluable to the broad scientific community. Detailed information about these resources is publicly available at the ENCODE portal (https://www.encodeproject.org/).


Assuntos
Bases de Dados Genéticas , Proteínas de Ligação a RNA/genética , RNA/metabolismo , Transcriptoma/genética , Sítios de Ligação , Humanos , Ligação Proteica , RNA/genética , RNA Interferente Pequeno/classificação , RNA Interferente Pequeno/genética , Proteínas de Ligação a RNA/metabolismo
7.
J Cell Physiol ; 232(7): 1689-1695, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27591397

RESUMO

Neutrophils from NOD (Non-Obese Diabetic) mice exhibited reduced migration speed, decreased frequency of directional changes, and loss of directionality during chemotaxis (compared to wild-type [WT] C57BL/6 mice). Additionally, F-actin of chemotaxing NOD neutrophils failed to orient toward the chemoattractant gradient and NOD neutrophil adhesion was impaired. A point mutation near the autophosphorylation site of Lyn in NOD mice was identified. Point mutations of G to A (G1412 in LynA and G1199 in LynB) cause a change of amino acid E393 (glutamic acid) to K (lysine) in LynA (E393 →K) (E372 of LynB), affecting fMLP-induced tyrosine phosphorylation. These data indicate that the Lyn mutation in NOD neutrophils is likely responsible for dysregulation of neutrophil adhesion and directed migration, implying the role of Lyn in modulating diabetic patient's susceptibility to bacterial and fungal infections. J. Cell. Physiol. 232: 1689-1695, 2017. © 2016 Wiley Periodicals, Inc.


Assuntos
Quimiotaxia , Mutação/genética , Neutrófilos/citologia , Neutrófilos/enzimologia , Quinases da Família src/metabolismo , Actinas/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Fibrinogênio/farmacologia , Fibronectinas/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Proteínas Mutantes/metabolismo , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Neutrófilos/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Fosfotirosina/metabolismo , Polimerização/efeitos dos fármacos
8.
Funct Integr Genomics ; 12(3): 501-14, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22038056

RESUMO

Several potential sources of reactive oxygen species (ROS) in cells exist. One source is NADPH oxidase, which is especially important for superoxide radical production. Nox2 is a primary regulatory subunit of NADPH oxidase. In the present study, we examined the role of ROS and NADPH oxidase in ischemic preconditioning (IP)-mediated cardioprotection by using Nox2(-/-) mice. Both wild-type (WT) and Nox2(-/-) mice were subjected to either 30 min of ischemia followed by 2 h of reperfusion (IR) or IP prior to 30 min ischemia and 2 h of reperfusion. Reduction in left ventricular developed pressure (60.1 versus 63 mmHg), dp/dt (max) (893 versus 1,027 mmHg/s), and aortic flow (0.9 versus 1.8 ml/min) was observed in Nox2(-/-)IPIR compared to WTIPIR along with increased infarct size (33% versus 22%) and apoptosis after 120 min of reperfusion. Differentially regulated genes were demonstrated by comparing gene expression in WTIPIR versus Nox2(-/-) IPIR hearts. Selected differentially regulated genes such as ß-catenin, SRPK3, ERDR1, ACIN1, Syntaxin-8, and STC1 were validated by real-time PCR. Taken together, this is the first report identifying important, differentially expressed genes during ischemic preconditioning in Nox2(-/-) mice by using microarray analysis.


Assuntos
Precondicionamento Isquêmico Miocárdico/métodos , Glicoproteínas de Membrana/metabolismo , Isquemia Miocárdica/enzimologia , NADPH Oxidases/metabolismo , Animais , Apoptose , Pressão Arterial , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Técnicas In Vitro , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Knockout , Isquemia Miocárdica/genética , Isquemia Miocárdica/patologia , Reperfusão Miocárdica/métodos , Miócitos Cardíacos/patologia , NADPH Oxidase 2 , NADPH Oxidases/genética , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Fatores de Tempo , Análise Serial de Tecidos , beta Catenina/genética , beta Catenina/metabolismo
9.
Toxicol Mech Methods ; 22(7): 555-9, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22694591

RESUMO

Oligonol (OLG), derived from lychee fruit, is a novel compound produced from the oligomerization of polyphenols. In this study, the acute effect of OLG treatment was investigated on heart, liver and kidney in rats. OLG treatment at two different doses (15 or 30 mg/kg body weight) and two different time points (1 day or 7 days of treatment) demonstrated that no toxic effects were observed on heart, liver and renal functions. Moreover, OLG did not induce any DNA damage or oxidative stress as measured by 8-hydroxy-2'-deoxyguanosine levels in plasma. OLG supplementation increased the phosphorylation of myocardial endothelial nitric oxide (NO) level (p-eNOS) in both the treatment groups. Even the low dose OLG treatment (15mg/kg b.w) demonstrated an increase in p-eNOS/eNOS ratio after normalization of p-eNOS values with eNOS on day 1 (1.5-fold) and day 7 (2.2-fold) groups as compared to control. The above results suggest that OLG treatment increases endothelial NO levels and may play a role in NO-mediated vasodilatory effects without adverse side effects on cardiovascular function. This endothelial NO production may underlie the beneficial effect of OLG in cardiovascular health.


Assuntos
Antioxidantes/efeitos adversos , Catequina/análogos & derivados , Coração/efeitos dos fármacos , Rim/efeitos dos fármacos , Litchi/química , Fígado/efeitos dos fármacos , Fenóis/efeitos adversos , Polifenóis/efeitos adversos , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Western Blotting , Catequina/efeitos adversos , Catequina/isolamento & purificação , Catequina/farmacologia , GMP Cíclico/sangue , Dano ao DNA , Relação Dose-Resposta a Droga , Ecocardiografia , Rim/enzimologia , Rim/metabolismo , Testes de Função Renal , Fígado/enzimologia , Fígado/metabolismo , Testes de Função Hepática , Masculino , Miocárdio/enzimologia , Miocárdio/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Fenóis/isolamento & purificação , Fenóis/farmacologia , Fosforilação , Polifenóis/isolamento & purificação , Polifenóis/farmacologia , Ratos , Ratos Wistar
10.
Ann Med Surg (Lond) ; 84: 104849, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36582861

RESUMO

In the treatment of some gynecological diseases, ureteral stents (double J stents) have been generally utilized in the prevention of ureteral injury. Nevertheless, if the ureteral stent is retained as a protective reminder during gynecological surgery, severe ureteral injury can be avoided. Hence, it is very essential to be familiar with the anatomy of ureter in gynecological surgery to prevent complications and morbidity. We demonstrate a case of a 49-year-old woman who presented with an electric burn broken ureteral stent in the gynecological surgery, but the ureter is only burned but not broken. This resulted in no abnormality being found during surgery. So, ureteroscopy is necessary to extract the ureteral stent and the patient is inserted with another new ureteral stent to repair the ureteral injury. The electric burn broken ureteral stent is difficult for discovering during the operation. And ureteroscopy is very crucial for the special situation of the ureteral injury.

11.
Urol Case Rep ; 44: 102148, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35846518

RESUMO

The stone inside the Allium bulbar urethral stent for treatment of urethral stenosis is an exceedingly rare disease. Herein, we report a case of the stone inside the Allium bulbar urethral stent(BUS) for treating urethral stricture in a 48-year-old Chinese male patient. The patient underwent a cystoscopy and URS for the stone inside BUS. The patient had only a symptom of urodynia. Urination of the patient is unobstructed after removing BUS and the urethral stricture of the patient was cured.

12.
J Mol Cell Cardiol ; 50(1): 239-47, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21074540

RESUMO

Oxidative stress plays a crucial role in disruption of neovascularization by alterations in thioredoxin 1 (Trx1) expression and its interaction with other proteins after myocardial infarction (MI). We previously showed that Trx1 has angiogenic properties, but the possible therapeutic significance of overexpressing Trx1 in chronic MI has not been elucidated. Therefore, we explored the angiogenic and cardioprotective potential of Trx1 in an in vivo MI model using transgenic mice overexpressing Trx1. Wild-type (W) and Trx1 transgenic (Trx1(Tg/+)) mice were randomized into W sham (WS), Trx1(Tg/+) sham (TS), WMI, and TMI. MI was induced by permanent occlusion of LAD coronary artery. Hearts from mice overexpressing Trx1 exhibited reduced fibrosis and oxidative stress and attenuated cardiomyocyte apoptosis along with increased vessel formation compared to WMI. We found significant inhibition of Trx1 regulating proteins, TXNIP and AKAP 12, and increased p-Akt, p-eNOS, p-GSK-3ß, HIF-1α, ß-catenin, VEGF, Bcl-2, and survivin expression in TMI compared to WMI. Echocardiography performed 30days after MI revealed significant improvement in myocardial functions in TMI compared to WMI. Our study identifies a potential role for Trx1 overexpression and its association with its regulatory proteins TXNIP, AKAP12, and subsequent activation of Akt/GSK-3ß/ß-catenin/HIF-1α-mediated VEGF and eNOS expression in inducing angiogenesis and reduced ventricular remodeling. Hence, Trx1 and other proteins identified in our study may prove to be potential therapeutic targets in the treatment of ischemic heart disease.


Assuntos
Infarto do Miocárdio/metabolismo , Neovascularização Patológica/metabolismo , Tiorredoxinas/metabolismo , Remodelação Ventricular/fisiologia , Proteínas de Ancoragem à Quinase A/metabolismo , Animais , Western Blotting , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/metabolismo , Ecocardiografia , Ensaio de Desvio de Mobilidade Eletroforética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Infarto do Miocárdio/genética , Neovascularização Patológica/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/fisiologia , Distribuição Aleatória , Tiorredoxinas/genética , Remodelação Ventricular/genética
13.
Circulation ; 121(10): 1244-55, 2010 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-20194885

RESUMO

BACKGROUND: The present study evaluated the reversal of diabetes-mediated impairment of angiogenesis in a myocardial infarction model of type 1 diabetic rats by intramyocardial administration of an adenoviral vector encoding thioredoxin-1 (Ad.Trx1). Various studies have linked diabetes-mediated impairment of angiogenesis to dysfunctional antioxidant systems in which thioredoxin-1 plays a central role. METHODS AND RESULTS: Ad.Trx1 was administered intramyocardially in nondiabetic and diabetic rats immediately after myocardial infarction. Ad.LacZ was similarly administered to the respective control groups. The hearts were excised for molecular and immunohistochemical analysis at predetermined time points. Myocardial function was measured by echocardiography 30 days after the intervention. The Ad.Trx1-administered group exhibited reduced fibrosis, oxidative stress, and cardiomyocyte and endothelial cell apoptosis compared with the diabetic myocardial infarction group, along with increased capillary and arteriolar density. Western blot and immunohistochemical analysis demonstrated myocardial overexpression of thioredoxin-1, heme oxygenase-1, vascular endothelial growth factor, and p38 mitogen-activated protein kinase-beta, as well as decreased phosphorylated JNK and p38 mitogen-activated protein kinase-alpha, in the Ad.Trx1-treated diabetic group. Conversely, we observed a significant reduction in the expression of vascular endothelial growth factor in nondiabetic and diabetic animals treated with tin protoporphyrin (SnPP, a heme oxygenase-1 enzyme inhibitor), even after Ad.Trx1 therapy. Echocardiographic analysis after 4 weeks of myocardial infarction revealed significant improvement in myocardial functional parameters such as ejection fraction, fractional shortening, and E/A ratio in the Ad.Trx1-administered group compared with the diabetic myocardial infarction group. CONCLUSIONS: This study demonstrates for the first time that impairment of angiogenesis and myocardial dysfunction can be regulated by Ad.Trx1 gene therapy in streptozotocin-induced diabetic rats subjected to infarction.


Assuntos
Diabetes Mellitus Experimental/complicações , Terapia Genética , Infarto do Miocárdio/terapia , Neovascularização Fisiológica , Transdução de Sinais , Tiorredoxinas/genética , Remodelação Ventricular , Animais , Apoptose , Células Cultivadas , Células Endoteliais/metabolismo , Fibrose , Heme Oxigenase-1/metabolismo , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/análise , Masculino , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Estreptozocina , Tiorredoxinas/análise , Transfecção , Fator A de Crescimento do Endotélio Vascular/análise , Proteínas Quinases p38 Ativadas por Mitógeno/análise
14.
Biochim Biophys Acta ; 1792(1): 39-48, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19027847

RESUMO

Diabetes, one of the major risk factors of metabolic syndrome culminates in the development of Ischemic Heart Disease (IHD). Refined diets that lack micronutrients, mainly trivalent chromium (Cr(3+)) have been identified as the contributor in the rising incidence of diabetes. We investigated the effect of niacin-bound chromium (NBC) during ischemia/reperfusion (IR) injury in streptozotocin induced diabetic rats. Rats were randomized into: Control (Con); Diabetic (Dia) and Diabetic rats fed with NBC (Dia+NBC). After 30 days of treatment, the isolated hearts were subjected to 30 min of global ischemia followed by 2 h of reperfusion. NBC treatment demonstrated significant increase in left ventricular functions and significant reduction in infarct size and cardiomyocyte apoptosis in Dia+NBC compared with Dia. Increased Glut-4 translocation to the lipid raft fractions was also observed in Dia+NBC compared to Dia. Reduced Cav-1 and increased Cav-3 expression along with phosphorylation of Akt, eNOS and AMPK might have resulted in increased Glut-4 translocation in Dia+NBC. Our results indicate that the cardioprotective effect of NBC is mediated by increased activation of AMPK, Akt and eNOS resulting in increased translocation of Glut-4 to the caveolar raft fractions thereby alleviating the effects of IR injury in the diabetic myocardium.


Assuntos
Cardiotônicos/farmacologia , Cromo/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Transportador de Glucose Tipo 4/metabolismo , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/metabolismo , Niacina/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Apoptose/efeitos dos fármacos , Transporte Biológico Ativo/efeitos dos fármacos , Cavéolas/efeitos dos fármacos , Cavéolas/metabolismo , Caveolinas/metabolismo , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/metabolismo , Técnicas In Vitro , Masculino , Traumatismo por Reperfusão Miocárdica/complicações , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/patologia , Óxido Nítrico Sintase Tipo III/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley
15.
Mol Cell Biochem ; 340(1-2): 73-80, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20213262

RESUMO

Calcium is an essential mineral to support bone health and serves as a major therapeutic intervention to prevent and delay the incidence of osteoporosis. Many individuals do not obtain the optimum amount of calcium from diets and depend on bioavailable calcium supplements. The present study was conducted to examine the effect of a novel plant-based calcium supplement, derived from marine algae, and contains high levels of calcium, magnesium, and other bone supporting minerals [commercially known as AlgaeCal (AC)], on proliferation, mineralization, and oxidative stress in cultured human osteoblast cells, and compared with inorganic calcium carbonate and calcium citrate salts. Cultured human fetal osteoblast cells (hFOB 1.19) were treated with AC (0.5 mg/ml, fixed by MTT assay), calcium carbonate, or calcium citrate. These cells were harvested after 4 days of treatment for ALP activity, PCNA expression, and DNA synthesis, and 2 days for Ca(2+) deposition in the presence and absence of vitamin D3 (5 nM). The ability of AC to reduce H(2)O(2) (0.3 mM)-induced oxidative stress was assessed after 24 h of treatment. ALP activity was significantly increased with AC treatment when compared to control, calcium carbonate, or calcium citrate (4.0-, 2.0-, and 2.5-fold, respectively). PCNA expression (immunocytochemical analysis), DNA synthesis (4.0-, 3.0-, and 4.0-fold, respectively), and Ca(2+) deposition (2.0-, 1.0-, and 4.0-fold, respectively) were significantly increased in AC-treated cells when compared with control, calcium carbonate, or calcium citrate treatment. These markers were further enhanced following additional supplementation of vitamin D3 in the AC-treated group cells. AC treatment significantly reduced the H(2)O(2)-induced oxidative stress when compared to calcium carbonate or calcium citrate (1.5- and 1.4-fold, respectively). These findings suggest that AC may serve as a superior calcium supplement as compared to other calcium salts tested in the present study. Hence, AC may be developed as a novel anti-osteoporotic supplement in the near future.


Assuntos
Calcificação Fisiológica/efeitos dos fármacos , Carbonato de Cálcio/farmacologia , Citrato de Cálcio/farmacologia , Proliferação de Células/efeitos dos fármacos , Suplementos Nutricionais , Eucariotos/química , Osteoblastos/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Linhagem Celular , Colecalciferol/farmacologia , Replicação do DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Peróxido de Hidrogênio/farmacologia , Osteoblastos/metabolismo , Oxidantes/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Fatores de Tempo
16.
Exp Ther Med ; 17(4): 2457-2464, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30906433

RESUMO

Kidney renal clear cell carcinoma (KIRC) is the most common type of renal cell carcinoma. While a number of treatments have been developed over the past few decades, the prognosis of patients with KIRC remains poor due to tumor metastasis and recurrence. Therefore, the molecular mechanisms of KIRC require to be elucidated in order to identify novel biomarkers. MicroRNAs (miRNAs/miRs) have been studied as important regulators of gene expression in a variety of cancer types. In the present study, a bioinformatics analysis of differentially expressed miRNAs in KIRC vs. normal tissues was performed based on raw miRNA expression data and patient information downloaded from the The Cancer Genome Atlas database. Furthermore, the clinical significance of differentially expressed miRNAs was evaluated, and their target genes and biological effects were further predicted. After applying the cut-off criteria of an absolute fold change of ≥2 and P<0.05, 127 differentially expressed miRNAs between KIRC and normal tissues were identified. The product of the miR-183/182/96 gene cluster, namely miR-183, miR-96 and miR-182, was revealed to be associated with multiple clinicopathological features of KIRC and to have a significant predictive and prognostic value. Subsequent functional enrichment analysis indicated that the target genes of the three miRNAs are associated with various Panther pathways, including the α-adrenergic receptor signaling pathway, metabotropic glutamate receptor group I pathway, histamine H1 receptor-mediated signaling pathway and thyrotropin-releasing hormone receptor signaling pathway. In addition, major enriched gene ontology terms in the category biological process included the intracellular signaling cascade, cellular macromolecule catabolic process and response to DNA damage stimulus. Taken together, the present study suggested that miR-183, miR-96 and miR-182 may function as potential carcinogenic factors in KIRC and may be utilized as prognostic predictors.

17.
Commun Biol ; 2: 19, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30652130

RESUMO

Adenosine-to-inosine (A-to-I) editing, mediated by the ADAR enzymes, diversifies the transcriptome by altering RNA sequences. Recent studies reported global changes in RNA editing in disease and development. Such widespread editing variations necessitate an improved understanding of the regulatory mechanisms of RNA editing. Here, we study the roles of >200 RNA-binding proteins (RBPs) in mediating RNA editing in two human cell lines. Using RNA-sequencing and global protein-RNA binding data, we identify a number of RBPs as key regulators of A-to-I editing. These RBPs, such as TDP-43, DROSHA, NF45/90 and Ro60, mediate editing through various mechanisms including regulation of ADAR1 expression, interaction with ADAR1, and binding to Alu elements. We highlight that editing regulation by Ro60 is consistent with the global up-regulation of RNA editing in systemic lupus erythematosus. Additionally, most key editing regulators act in a cell type-specific manner. Together, our work provides insights for the regulatory mechanisms of RNA editing.


Assuntos
Adenosina Desaminase/genética , Adenosina Desaminase/metabolismo , Regulação Neoplásica da Expressão Gênica , Edição de RNA/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Adenosina/genética , Elementos Alu , Autoantígenos/genética , Técnicas de Silenciamento de Genes , Células Hep G2 , Humanos , Inosina/genética , Células K562 , Lúpus Eritematoso Sistêmico/genética , RNA Citoplasmático Pequeno/genética , Ribonucleoproteínas/genética , Análise de Sequência de RNA , Transcrição Gênica , Transfecção
19.
J Mol Cell Cardiol ; 44(1): 170-9, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18001768

RESUMO

Hypercholesterolemia (HC) induced endothelial cell dysfunction and decreased endothelial nitric oxide formation results in impaired angiogenesis and subsequent cardiovascular disorders. Therapeutic angiogenesis is known to be a novel strategy for treatment of patients with ischemic heart disease. We have shown that secoisolariciresinol diglucoside (SDG) is angiogenic as well as cardioprotective against myocardial ischemia. In the present study, we examined the efficacy of SDG in a hypercholesterolemic myocardial infarction (MI) model. The rats were maintained on a normal and high cholesterol diet (2%) for 8 weeks followed by oral administration of SDG (20 mg/kg) for 2 weeks. The rats were divided into four groups (n=24 in each): Control (C); SDG control (SDG); HC; and HC+SDG (HSDG). Isolated hearts subjected to 30 min of global ischemia followed by 120 min of reperfusion were used to measure the cardiac functions, infarct size and to examine the protein expression profile. After treatment, MI was induced by ligating the left anterior descending artery. Echocardiographic parameters were examined 30 days after MI. Significant reduction in total cholesterol, LDL-cholesterol, triglycerides and an increase in HDL-cholesterol levels were observed in HSDG as compared to the HC. Decreased infarct size was observed in the HSDG group (43%) compared to the HC (54%). Increased phosphorylation of endothelial nitric oxide synthase (p-eNOS) (3.1-fold), vascular endothelial growth factor (1.9-fold) and heme oxygenase-1 (2.3-fold) was observed in the HSDG group as compared to the HC group. Significant improvement in left ventricular functions was also observed in the HSDG group as evidenced by increased ejection fraction (55% vs. 45%), fractional shortening (28% vs. 22%) and decreased left ventricular inner diameter in systole (8 vs. 6 mm) in HSDG compared to HC. Moreover, MI model has shown increased capillary density (2531 vs. 1901) and arteriolar density (2.6 vs. 1.8) in SDG-treated rats as compared to the HC. The increased capillary and arteriolar density along with increased left ventricular functions on SDG treatment might be due to increased HO-1, VEGF and p-eNOS expression. In conclusion, our study demonstrates for the first time that SDG treatment reduces ventricular remodeling by neovascularization of the infarcted HC myocardium.


Assuntos
Butileno Glicóis/farmacologia , Cardiotônicos/farmacologia , Glucosídeos/farmacologia , Hipercolesterolemia/complicações , Traumatismo por Reperfusão Miocárdica/complicações , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/patologia , Neovascularização Fisiológica/efeitos dos fármacos , Animais , Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/metabolismo , Western Blotting , Vasos Coronários/efeitos dos fármacos , Testes de Função Cardíaca , Heme Oxigenase-1/metabolismo , Hipercolesterolemia/enzimologia , Hipercolesterolemia/fisiopatologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Infarto do Miocárdio/diagnóstico por imagem , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/enzimologia , Miocárdio/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Fosfoproteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Ultrassonografia , Fator A de Crescimento do Endotélio Vascular/metabolismo
20.
Genome Biol ; 19(1): 71, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29859120

RESUMO

Understanding the functional impact of genomic variants is a major goal of modern genetics and personalized medicine. Although many synonymous and non-coding variants act through altering the efficiency of pre-mRNA splicing, it is difficult to predict how these variants impact pre-mRNA splicing. Here, we describe a massively parallel approach we use to test the impact on pre-mRNA splicing of 2059 human genetic variants spanning 110 alternative exons. This method, called variant exon sequencing (Vex-seq), yields data that reinforce known mechanisms of pre-mRNA splicing, identifies variants that impact pre-mRNA splicing, and will be useful for increasing our understanding of genome function.


Assuntos
Processamento Alternativo , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Precursores de RNA/metabolismo , RNA Mensageiro/metabolismo , Análise de Sequência de RNA/métodos , Éxons , Humanos , Íntrons , Sítios de Splice de RNA
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA