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1.
Chinese Journal of Biotechnology ; (12): 2595-2602, 2021.
Article in Chinese | WPRIM | ID: wpr-887825

ABSTRACT

Nuclear receptor subfamily 2, group F, member 6 (NR2F6) is a member of orphan nuclear receptors, which is expressed in major tissues and organs of the human body, and plays an important role in the regulation of various biological functions and gene expressions. Recent studies have shown that the expression of NR2F6 was up-regulated in a variety of malignant tumors and showed significant correlations with cancer progression. These findings triggered the widespread interest in understanding the relationship between NR2F6 and cancer development and progression. In addition, the latest studies have underscored that NR2F6 was involved in enhancing antitumor immune responses that could serve as a potential target for immune regulation. This review summarizes the biological functions of NR2F6 and its role in tumors, with the aim to provide new insights into effective cancer therapies.


Subject(s)
Gene Expression Regulation , Humans , Neoplasms/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Repressor Proteins/genetics , Transcription Factors/genetics
2.
Acta Physiologica Sinica ; (6): 795-804, 2021.
Article in English | WPRIM | ID: wpr-921282

ABSTRACT

Farnesoid X receptor (FXR) has been identified as an inhibitor of platelet function and an inducer of fibrinogen protein complex. However, the regulatory mechanism of FXR in hemostatic system remains incompletely understood. In this study, we aimed to investigate the functions of FXR in regulating antithrombin III (AT III). C57BL/6 mice and FXR knockout (FXR KO) mice were treated with or without GW4064 (30 mg/kg per day). FXR activation significantly prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT), lowered activity of activated factor X (FXa) and concentrations of thrombin-antithrombin complex (TAT) and activated factor II (FIIa), and increased level of AT III, whereas all of these effects were markedly reversed in FXR KO mice. In vivo, hepatic AT III mRNA and protein expression levels were up-regulated in wild-type mice after FXR activation, but down-regulated in FXR KO mice. In vitro study showed that FXR activation induced, while FXR knockdown inhibited, AT III expression in mouse primary hepatocytes. The luciferase assay and ChIP assay revealed that FXR can bind to the promoter region of AT III gene where FXR activation increased AT III transcription. These results suggest FXR activation inhibits coagulation process via inducing hepatic AT III expression in mice. The present study reveals a new role of FXR in hemostatic homeostasis and indicates that FXR might act as a potential therapeutic target for diseases related to hypercoagulation.


Subject(s)
Animals , Antithrombin III , Blood Coagulation , Hepatocytes , Liver , Mice , Mice, Inbred C57BL , Mice, Knockout , Receptors, Cytoplasmic and Nuclear/genetics
3.
Braz. j. med. biol. res ; 51(7): e7312, 2018. tab, graf
Article in English | LILACS | ID: biblio-951734

ABSTRACT

Farnesoid X receptor (FXR) and related pathways are involved in the therapeutic effect of sleeve gastrectomy for overweight or obese patients with diabetes mellitus. This study aimed to investigate the mechanism of FXR expression regulation during the surgical treatment of obese diabetes mellitus by sleeve gastrectomy. Diabetic rats were established by combined streptozotocin and high-fat diet induction. Data collection included body weight, chemical indexes of glucose and lipid metabolism, liver function, and the expression levels of musculoaponeurotic fibrosarcoma oncogene family B (MAFB), FXR, and related genes induced by sleeve gastrectomy. Chang liver cells overexpressing MAFB gene were established to confirm the expression of related genes. The binding and activation of FXR gene by MAFB were tested by Chip and luciferase reporter gene assays. Vertical sleeve gastrectomy induced significant weight loss and decreased blood glucose and lipids in diabetic rat livers, as well as decreased lipid deposition and recovered lipid function. The expression of MAFB, FXR, and FXR-regulated genes in diabetic rat livers were also restored by sleeve gastrectomy. Overexpression of MAFB in Chang liver cells led to FXR gene expression activation and the alteration of multiple FXR-regulated genes. Chip assay showed that MAFB could directly bind with FXR promoter, and the activation of FXR expression was confirmed by luciferase reporter gene analysis. The therapeutic effect of sleeve gastrectomy for overweight or obese patients with diabetes mellitus was mediated by activation of FXR expression through the binding of MAFB transcription factor.


Subject(s)
Animals , Male , Rats , Oncogene Proteins/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , Diabetes Mellitus, Experimental/metabolism , MafB Transcription Factor/metabolism , Gastrectomy/methods , Obesity/surgery , Gene Expression Regulation , Blotting, Western , Rats, Sprague-Dawley , Oncogene Proteins/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Reverse Transcriptase Polymerase Chain Reaction , MafB Transcription Factor/genetics , Lipid Metabolism , Liver/metabolism , Obesity/metabolism
5.
Arq. bras. endocrinol. metab ; 57(7): 513-519, out. 2013. tab
Article in English | LILACS | ID: lil-690588

ABSTRACT

OBJECTIVE: The aim of the present study was investigate the association between six genetic variants in the nuclear receptor genes PPARA, RXRA, NR1I2 and NR1I3 and the lipid-lowering efficacy and safety of statin therapy. SUBJECTS AND METHODS: The study was carried out on 240 Brazilian hypercholesterolemic patients on simvastatin and atorvastatin therapy. The polymorphisms were analyzed by PCR-based methods. RESULTS: The NR1I3 rs2307424 genotype distribution was different between subjects with and without adverse drug reactions. Among subjects in the ADR group, no T/T homozygotes were observed for this polymorphism, while in the non-ADR group the frequency of this genotype was 19.4% (P = 0.007, after multiple testing corrections P = 0.042). CONCLUSION: The polymorphisms investigated in PPARA (rs1800206), RXRA (rs11381416), and NR1I2 (rs1523130) did not influence the lipid-lowering efficacy and safety of statin. Our results show the possible influence of NR1I3 genetic variant on the safety of statin.


OBJETIVO: O objetivo deste estudo foi investigar a associação de seis variantes genéticas nos genes de receptores nucleares PPARA, RXRA, NR1I2 e NR1I3 na eficácia hipolipemiante e na segurança da terapia com estatinas. SUJEITOS E MÉTODOS: O estudo foi realizado com 240 pacientes hipercolesterolêmicos em terapia com sinvastina e atorvastatina. Os polimorfismos foram analisados por meio de métodos baseados em PCR. RESULTADOS: A distribuição da frequência genotípica do polimorfismo NR1I3 rs2307424 foi diferente entre os pacientes com e sem efeito adverso à medicação; entre os sujeitos do grupo com efeitos adversos, nenhum homozigoto T/T foi observado, enquanto no grupo de indivíduos sem efeitos adversos a frequência desse genótipo foi 19,4% (P = 0,007, após correção para múltiplos testes P = 0,042). CONCLUSÃO: Os polimorfismos investigados nos genes PPARA (rs1800206), RXRA (rs11381416) e NR1I2 (rs1523130) não foram associados com eficácia hipolipemiante e segurança da terapia com estatinas. Nossos resultados mostram uma possível influência de variantes do gene NR1I3 (rs2307424) no desenvolvimento de efeitos adversos à terapia com estatinas.


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Anticholesteremic Agents/therapeutic use , Dyslipidemias/drug therapy , Polymorphism, Genetic , PPAR alpha/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Steroid/genetics , Retinoid X Receptor alpha/genetics , Alleles , Anticholesteremic Agents/adverse effects , Dyslipidemias/genetics , Genotype , Heptanoic Acids/adverse effects , Heptanoic Acids/therapeutic use , Lipids/blood , Polymerase Chain Reaction , Pyrroles/adverse effects , Pyrroles/therapeutic use , Risk Factors , Simvastatin/adverse effects , Simvastatin/therapeutic use , Treatment Outcome
6.
Braz. j. med. biol. res ; 44(7): 642-646, July 2011. ilus, tab
Article in English | LILACS | ID: lil-595708

ABSTRACT

To explore how cytohesin-1 (CYTH-1) small interfering RNA (siRNA) influences the insulin-like growth factor receptor (IGFR)-associated signal transduction in prostate cancer, we transfected human prostate cancer PC-3 cell lines with liposome-encapsulatedCYTH-1 siRNA in serum-free medium and exposed the cells to 100 nM IGF-1. The mRNA and protein levels of the signal molecules involved in the IGFR signaling pathways were determined by real-time PCR and detected by Western blotting. The relative mRNA levels of CYTH-1, c-Myc, cyclinD1 and IGF-1R (CYTH-1 siRNA group vs scrambled siRNA group) were 0.26 vs 0.97, 0.34 vs 1.06, 0.10 vs 0.95, and 0.27 vs 0.41 (P < 0.05 for all), respectively. The relative protein levels of CYTH-1, pIGF-1R, pIRS1, pAkt1, pErk1, c-Myc, and cyclinD1 (CYTH-1 siRNA group vsscrambled siRNA group) were 0.10 vs 1.00 (30 min), 0.10 vs 0.98 (30 min), 0.04 vs 0.50 (30 min), 0.10 vs 1.00 (30 min), 0.10 vs 1.00 (30 min), 0.13 vs 0.85 (5 h), and 0.08 vs 0.80 (7 h), respectively. The tyrosine kinase activity of IGF-1R was associated with CYTH-1. The proliferative activity of PC-3 cells transfected with CYTH-1 siRNA was significantly lower than that of cells transfected with scrambled siRNA at 48 h (40.5 vs87.6 percent, P < 0.05) and at 72 h (34.5 vs 93.5 percent, P < 0.05). In conclusion, the interference of siRNA with cytohesin-1 leads to reduced IGFR signaling in prostate cancer; therefore, CYTH-1 might serve as a new molecular target for the treatment of prostate cancer.


Subject(s)
Humans , Male , Guanine Nucleotide Exchange Factors/antagonists & inhibitors , Prostatic Neoplasms/metabolism , RNA, Small Interfering/pharmacology , Receptors, Somatomedin/metabolism , Signal Transduction/drug effects , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , GTPase-Activating Proteins/genetics , GTPase-Activating Proteins/metabolism , Guanine Nucleotide Exchange Factors/genetics , Insulin-Like Growth Factor I/metabolism , Phosphorylation , Prostatic Neoplasms/pathology , Real-Time Polymerase Chain Reaction , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism
7.
Article in English | WPRIM | ID: wpr-131296

ABSTRACT

Cholesterol 7alpha-hydroxylase (CYP7A1) regulates the balance between cholesterol supply and metabolism by catalyzing the rate-limiting step of bile acid biosynthesis. The transcriptional activity of CYP7A1 is tightly controlled by various nuclear receptors. A forkhead transcription factor O1 (FOXO1) plays a critical role in metabolism, and insulin inactivates FOXO1 through Akt-dependent phosphorylation and nuclear exclusion. We investigated the role of insulin-Akt-FOXO1 signaling pathway in CYP7A1 transcriptional regulation since we found putative insulin-response elements, FOXO1 binding sequences, in both rat and human CYP7A1 promoters. However, ectopic expression of FOXO1 increased the rat CYP7A1-, but mildly reduced human CYP7A1-promoter activities in a dose-dependent manner. Similarly to bile acids, insulin treatment increased small heterodimer partner (SHP) mRNA rapidly and transiently, leading to the suppression of CYP7A1 transcription in both human and rodents. Chromatin immunoprecipitation showed that FOXO1 directly bound to rat CYP1A1 promoter in the absence of insulin. FOXO1 binding to the rat promoter was diminished by insulin treatment as well as by expression of SHP. Our results suggest that the stimulation of insulin- signaling pathway of Akt-FOXO1 and SHP expression may regulate cholesterol/bile acid metabolisms in liver, linking carbohydrate and cholesterol metabolic pathways. A prolonged exposure of insulin in hyperinsulinemic insulin resistance or diabetic status represses CYP7A1 transcription and bile acid biosynthesis through SHP induction and FOXO1 inactivation, leading to impairment of the hepatic cholesterol/bile acid metabolisms.


Subject(s)
Animals , Bile Acids and Salts/metabolism , Cholesterol/metabolism , Cholesterol 7-alpha-Hydroxylase/genetics , Forkhead Transcription Factors/genetics , Gene Expression Regulation/drug effects , Glucose/metabolism , Hep G2 Cells , Humans , Insulin/pharmacology , Lipid Metabolism/drug effects , Liver/metabolism , Mice , Mice, Inbred C57BL , Mutagenesis, Site-Directed , Nerve Tissue Proteins/genetics , Protein Binding/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Rats , Receptors, Cytoplasmic and Nuclear/genetics , Sequence Deletion/genetics , Signal Transduction/drug effects , Transcriptional Activation/drug effects
8.
Article in English | WPRIM | ID: wpr-131293

ABSTRACT

Cholesterol 7alpha-hydroxylase (CYP7A1) regulates the balance between cholesterol supply and metabolism by catalyzing the rate-limiting step of bile acid biosynthesis. The transcriptional activity of CYP7A1 is tightly controlled by various nuclear receptors. A forkhead transcription factor O1 (FOXO1) plays a critical role in metabolism, and insulin inactivates FOXO1 through Akt-dependent phosphorylation and nuclear exclusion. We investigated the role of insulin-Akt-FOXO1 signaling pathway in CYP7A1 transcriptional regulation since we found putative insulin-response elements, FOXO1 binding sequences, in both rat and human CYP7A1 promoters. However, ectopic expression of FOXO1 increased the rat CYP7A1-, but mildly reduced human CYP7A1-promoter activities in a dose-dependent manner. Similarly to bile acids, insulin treatment increased small heterodimer partner (SHP) mRNA rapidly and transiently, leading to the suppression of CYP7A1 transcription in both human and rodents. Chromatin immunoprecipitation showed that FOXO1 directly bound to rat CYP1A1 promoter in the absence of insulin. FOXO1 binding to the rat promoter was diminished by insulin treatment as well as by expression of SHP. Our results suggest that the stimulation of insulin- signaling pathway of Akt-FOXO1 and SHP expression may regulate cholesterol/bile acid metabolisms in liver, linking carbohydrate and cholesterol metabolic pathways. A prolonged exposure of insulin in hyperinsulinemic insulin resistance or diabetic status represses CYP7A1 transcription and bile acid biosynthesis through SHP induction and FOXO1 inactivation, leading to impairment of the hepatic cholesterol/bile acid metabolisms.


Subject(s)
Animals , Bile Acids and Salts/metabolism , Cholesterol/metabolism , Cholesterol 7-alpha-Hydroxylase/genetics , Forkhead Transcription Factors/genetics , Gene Expression Regulation/drug effects , Glucose/metabolism , Hep G2 Cells , Humans , Insulin/pharmacology , Lipid Metabolism/drug effects , Liver/metabolism , Mice , Mice, Inbred C57BL , Mutagenesis, Site-Directed , Nerve Tissue Proteins/genetics , Protein Binding/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Rats , Receptors, Cytoplasmic and Nuclear/genetics , Sequence Deletion/genetics , Signal Transduction/drug effects , Transcriptional Activation/drug effects
9.
Article in English | WPRIM | ID: wpr-104282

ABSTRACT

Angiotensin II is a major effector molecule in the development of cardiovascular disease. In vascular smooth muscle cells (VSMCs), angiotensin II promotes cellular proliferation and extracellular matrix accumulation through the upregulation of plasminogen activator inhibitor-1 (PAI-1) expression. Previously, we demonstrated that small heterodimer partner (SHP) represses PAI-1 expression in the liver through the inhibition of TGF-beta signaling pathways. Here, we investigated whether SHP inhibited angiotensin II-stimulated PAI-1 expression in VSMCs. Adenovirus-mediated overexpression of SHP (Ad-SHP) in VSMCs inhibited angiotensin II- and TGF-beta-stimulated PAI-1 expression. Ad-SHP also inhibited angiotensin II-, TGF-beta- and Smad3-stimulated PAI-1 promoter activity, and angiotensin II-stimulated AP-1 activity. The level of PAI-1 expression was significantly higher in VSMCs of SHP-/- mice than wild type mice. Moreover, loss of SHP increased PAI-1 mRNA expression after angiotensin II treatment. These results suggest that SHP inhibits PAI-1 expression in VSMCs through the suppression of TGF-beta/Smad3 and AP-1 activity. Thus, agents that target the induction of SHP expression in VSMCs might help prevent the development and progression of atherosclerosis.


Subject(s)
Adenoviridae/genetics , Angiotensin II/pharmacology , Animals , Blotting, Northern , Cells, Cultured , Electrophoretic Mobility Shift Assay , Genetic Vectors/genetics , Humans , Mice , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/drug effects , Plasminogen Activator Inhibitor 1/genetics , Promoter Regions, Genetic/genetics , Rats , Receptors, Cytoplasmic and Nuclear/genetics , Reverse Transcriptase Polymerase Chain Reaction , Smad3 Protein/genetics , Transforming Growth Factor beta/pharmacology
10.
Article in English | WPRIM | ID: wpr-196694

ABSTRACT

Small heterodimer partner (SHP) is an atypical member of nuclear receptor superfamily that lacks a DNA-binding domain. In previous study, we showed that SHP, c-jun, p65 of NF-kappaB subunits, and p21WAF1 expression was increased during monocytic differentiaton with the exposure of human leukemia cells to a differentiation agent, PMA. In this study, c-Jun and p65 were shown to mediate the transcriptional activation of the SHP promoter. In addition, SHP induced the cell cycle regulatory protein levels and cooperatively increased an induction of p21WAF1 expression with p65. Furthermore, SHP protected differentiated cells from etoposide-induced cellular apoptosis through the induction and cytoplasmic sequestration of p21WAF1. Complex formation between SHP and p21WAF1 was demonstrated by means of coimmunoprecipitation. These results suggest that SHP prolongs a cellular survival of differentiating monocytes through the transcriptional regulation of target genes of cell survival and differentiation.


Subject(s)
Apoptosis , Cell Differentiation , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p21/genetics , Gene Expression Regulation , Humans , Monocytes/cytology , Promoter Regions, Genetic , Proto-Oncogene Proteins c-jun/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Transcription Factor RelA/genetics
11.
Article in English | WPRIM | ID: wpr-96569

ABSTRACT

Oxidative stress plays critical roles in airway inflammation that is usually accompanied by increased vascular permeability and plasma exudation. VEGF increases vascular permeability and leads to airway inflammation. In addition, VEGF has been shown to enhance receptor activator of NF-kappaB (RANK) expression in endothelial cells. An aim of the study was to determine the potential role of antioxidant in the regulation of RANK expression in murine model of asthma. We have used a C57BL/6 mouse model of allergic asthma to evaluate the effect of L-2-oxothiazolidine-4-carboxylic acid (OTC), a prodrug of cysteine, which acts as an antioxidant, and VEGF receptor inhibitor on RANK mRNA expression. The mice develop the following pathophysiological features of asthma in the lungs: increased expression of RANK mRNA, increased number of inflammatory cells of the airways, increased vascular permeability, and increased levels of VEGF. Administration of OTC and VEGF receptor inhibitor markedly reduced plasma extravasation and VEGF levels in allergen-induced asthmatic lungs. We also showed that the increased RANK mRNA expression at 72 h after ovalbumin inhalation were reduced by the administration of OTC or VEGF receptor inhibitor. The results indicate that OTC and VEGF receptor inhibitor which inhibit up-regulation of VEGF expression modulate RANK expression that may be in association with the regulation of vascular permeability, and suggest that VEGF may regulate the RANK expression. These findings provide a crucial molecular mechanism for the potential use of antioxidants to prevent and/or treat asthma and other airway inflammatory disorders.


Subject(s)
Vascular Endothelial Growth Factor A/analysis , Thiazolidines , Thiazoles/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors , Receptors, Tumor Necrosis Factor/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Reactive Oxygen Species/metabolism , RNA, Messenger/genetics , Pyrrolidonecarboxylic Acid , Proto-Oncogene Proteins c-akt/metabolism , Protein Kinase Inhibitors/pharmacology , Prodrugs/pharmacology , Phosphorylation/drug effects , Ovalbumin/immunology , Osteoprotegerin , Mice, Inbred C57BL , Mice , Immunohistochemistry , Glycoproteins/genetics , Gene Expression/drug effects , Female , Capillary Permeability/drug effects , Bronchoalveolar Lavage Fluid/chemistry , Blotting, Western , Asthma/drug therapy , Antioxidants/pharmacology , Animals
12.
Article in English | WPRIM | ID: wpr-53152

ABSTRACT

To develop an inducible expression system, the enhanced artificial nuclear receptors and target reporters were constructed. Artificial nuclear receptors were generated by fusing three domains, consisting of DNA-binding domain (DBD) of GAL4, ligand binding domain (LBD) of progesterone or estrogen receptor, and activation domain (AD) of VP16, sterol regulatory element binding protein (SREBP)-1a, or SREBP-2. The activation domain of SREBP-1a showed most potent transcriptional activity. The maximal level of target reporter gene expression was extremely elevated by the usage of ATP citrate-lyase (ACL) minimal promoter -60/+67 in place of artificial TATA promoter, while the SV40 enhancer severely increased the basal transcription in the absence of ligand. The induction system, developed in the present study, was applied to cell therapy, resulting in successful induction of single-chain insulin analogue (SIA) gene expression to correct the hyperglycemia in diabetic animals. By means of subcutaneous cell therapy, the SIA gene expression rapidly occurred after the local topical application of ligand. These results suggest that our system represents a powerful tool for transcriptional regulation of target gene that can be used for diverse applications, ranging from basic research to gene therapy.


Subject(s)
Transfection , Transcriptional Activation , Receptors, Cytoplasmic and Nuclear/genetics , Mice, Inbred BALB C , Mice , Male , Ligands , Genetic Vectors/chemical synthesis , Genes, Reporter , Genetic Therapy/methods , Gene Expression Regulation , Diabetes Mellitus, Type 1/therapy , Diabetes Mellitus, Experimental/blood , Blood Glucose/analysis , Animals
13.
Article in English | WPRIM | ID: wpr-207073

ABSTRACT

TCR signaling leading to thymocyte apoptosis is mediated through the expression of the Nur77 family of orphan nuclear receptors. It has been shown that the Nur77 promoter is activated by at least two signaling pathways, one mediated by calcium and the other by protein kinase C (PKC). MEF2D has been known to regulate Nur77 expression in a calcium- dependent manner. The mechanism by which calcium regulates MEF2D is through dissociation of calcium-sensitive MEF2 corepressors (Cabin1/ HDACs, HDAC4/5) and the association with calcineurin-activated transcription factor NF-AT and the coactivator p300. However, little is known about how PKC activates the Nur77 promoter. Herein, we report that PKC theta targets AP-1 like response element in the Nur77 promoter where JunD constitutively binds. PKC theta triggers mitogen-activated protein kinase- inediated phosphorylation of JunD, and increases transcriptional activity of JunD, cooperatively with p300. Menin is identified as the transcriptional corepressor for JunD via recruitment of mSin3-istone deacetylases. In fact, Menin represses PKC theta/ p300-mediated transcriptional activity of JunD in T cell. Its dynamic regulation of histone modifiers with JunD is responsible for PKCq-synergistic effect on Nur77 expression in T cell.


Subject(s)
Cell Line, Tumor , DNA-Binding Proteins/genetics , Enzyme Activation , Gene Expression Regulation , Humans , Isoenzymes/metabolism , Mitogen-Activated Protein Kinases/metabolism , Multiple Endocrine Neoplasia Type 1 , Promoter Regions, Genetic/genetics , Protein Kinase C/metabolism , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins c-jun/antagonists & inhibitors , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Steroid/genetics , Response Elements , Transcription Factors/genetics , Transcription, Genetic/genetics
15.
São Paulo; s.n; 2004. [117] p. ilus, tab, graf.
Thesis in Portuguese | LILACS | ID: lil-397838

ABSTRACT

Com o objetivo de estudar a participação dos genes SF1 e DAX1 na etiologia dos distúrbios da determinação gonadal, analisamos uma casuística de 35 pacientes com sexo reverso 46,XY e 12 pacientes com sexo reverso 46,XX.. Estes pacientes foram estudados do ponto de vista clínico, hormonal, radiológico, anatomopatológico e molecular. Identificamos o primeiro paciente com uma mutação no gene SF1 associada a sexo reverso 46,XY sem insuficiência adrenal, e descrevemos o seu comportamento funcional in vitro. Identificamos também 9 variantes alélicas neste gene altamente conservado. /The aim of this study is to elucidate the participation of SF1 and DAX1 genes in the etiology of abnormal gonadal determination by analyzing 35 patients with 46,XY sex reversal and twelve 46,XX sex-reversed patients. All patients underwent clinical hormonal, radiological, anatomopathological and molecular evaluation. We identified the first case with a SF1 mutation associated to 46, XY sex reversal without adrenal insufficiency and characterized its transcriptional activity in vitro. We also identified 9 allelic variants in this highly conservative gene...


Subject(s)
Humans , Disorders of Sex Development , Sex Determination Processes , Disorders of Sex Development/diagnosis , Disorders of Sex Development/genetics , Receptors, Cytoplasmic and Nuclear/physiology , Receptors, Cytoplasmic and Nuclear/genetics
16.
Article in English | WPRIM | ID: wpr-171357

ABSTRACT

TNF-alpha, a trimeric cytokine, was known to inhibit differention of preadipocytes to adipocytes. In the present study, we investigated signal mediators working downstream of TNF-alpha using murine 3T3-L1 cells. TNF-alpha induced activation of both c-jun NH2-terminal kinase (JNK) and nuclear transcription factor-kappaB (NF-kappaB) in 3T3-L1 cells. Blockage of these two mediators activities by specific inhibitors, SP600125 and Ad-IkappaBalpha-SR restored adipogenesis differentiation suggesting their involvement in the inhibited differentiation of 3T3-L1 cells by TNF-alpha. Consistent with previous studies, peroxisome proliferator-activated receptor gamma (PPARgamma) a key transcriptional regulator was remarkably reduced by TNF-alpha treatment. Compared with adipogenesis, however, SP600125, a chemical JNK inhibitor hardly relieved TNF-alpha effect on PPARgamma expression whereas S32A/S36A mutant of IkappaBalpha considerably recovered PPARgamma expression, indicating that two signal mediators exploit separable main routes to achieve reduced adipogenesis. These results suggest that inhibition of 3T3-L1 cells differentiation by TNF-alpha is partly implemented through NF-kappaB and one of its downstream effectors be PPARgamma.


Subject(s)
3T3-L1 Cells , Adipocytes/cytology , Animals , Cell Differentiation/drug effects , Gene Expression Regulation/drug effects , Mice , NF-kappa B/metabolism , Promoter Regions, Genetic/genetics , RNA, Messenger/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Transcription Factors/genetics , Tumor Necrosis Factor-alpha/pharmacology
17.
Article in English | WPRIM | ID: wpr-127187

ABSTRACT

Information on precise effects of deflazacort on bone cell function, especially osteoclasts, is quite limited. Therefore, the present study was undertaken to test effects of deflazacort on osteoclast-like cell formation in mouse bone marrow cultures and on the regulation of osteoprotegerin (OPG) and its ligand (RANKL) mRNA expressions by RT-PCR in the ST2 marrow stromal cells. TRAP-positive mononuclear cells increased after the treatment of deflazacort at 10(-9) to 10(-7) M alone for 6 days in a dose-dependent manner. Number of TRAP-positive multi-nucleated cells (MNCs) increased significantly with combined treatment of deflazacort at 10(-7) M and 1,25-(OH)2D3 at 10(-9) M compared to that of cultures treated with 1,25-(OH)2D3 alone (p<0.05). Exposure to deflazacort at 10(-7) M in the presence of 1,25-(OH)2D3 at 10(-9) M in the last 3-day culture had greater stimulatory effect on osteoclast-like cell formation than that of the first 3-day culture did. Deflazacort at 10(-10) -10(-6) M downregulated OPG and upregulated RANKL in mRNA levels in a dose-dependent manner. These observations suggest that deflazacort stimulate osteoclast precursor in the absence of 1,25-(OH)2D3 and enhance differentiation of osteoclasts in the presence of 1,25-(OH)2D3. These effects are, in part, thought to be mediated by the regulation of the expression of OPG and RANKL mRNA in marrow stromal cells.


Subject(s)
Animals , Bone Marrow Cells/cytology , Calcitriol/pharmacology , Calcium Channel Agonists/pharmacology , Carrier Proteins/genetics , Cell Differentiation/drug effects , Cells, Cultured , Dexamethasone/pharmacology , Gene Expression/drug effects , Glucocorticoids/pharmacology , Glycoproteins/genetics , Immunosuppressive Agents/pharmacology , Male , Membrane Glycoproteins/genetics , Mice , Mice, Inbred ICR , Osteoclasts/cytology , Pregnenediones/pharmacology , RNA, Messenger/analysis , Receptors, Cytoplasmic and Nuclear/genetics , Stromal Cells/cytology
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