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1.
Toxicol Lett ; 318: 1-11, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31618665

RESUMO

Triptolide (TP), a principal bioactive component extracted from traditional Chinese medicine Tripterygium wilfordii Hook. F. (TWHF), has attracted wide attention of its therapeutic effects on inflammation and autoimmune diseases. However, the therapeutic application of TP is hindered by severe cardiomyocyte toxicity and narrow therapeutic window. We previously identified that the p53 was an indispensable contributor in TP-induced myocardial injury. p53 has an inhibitory effect on IKKß-NF-κB pathway that regulates glucose transporters (GLUT) expression. Based on these evidences, we speculate that p53 mediates TP-disturbed glucose uptake by blocking IKKß-NF-κB signaling. This study focused on the effect of TP on cardiac glucose uptake and the role of p53 in glucose metabolism in cardiomyocytes, and p53 -/- mice. TP treatment depressed glucose consumption and ATP production resulting in myocardial damage. Incubation with ATP (5 mM) remarkably decreased the cellular damage. Immunoblotting and immunofluorescence identified that TP suppressed glucose uptake by restricting IKKß-NF-κB signaling activation, GLUT1 and GLUT4 expression. p53 inhibition alleviated the cell damage and the compromise of glucose uptake. Mechanistically, p53 antagonist PFTα abolished TP-induced the inhibition of IKKß, IκBα phosphorylation, p65 nuclear translocation, and GLUT1, GLUT4 expression. Consistently, in acute heart injury models, p53 deficiency upregulated IKKß-NF-κB activation and GLUT1, GLUT4 protein levels which was also indicated as amelioration of heart histological injury after 1.2 mg kg-1 TP administration. The present findings indicate that TP-induced p53 overactivation suppresses glucose uptake by inhibiting IKKß-NF-κB pathway and downregulating NF-κB-dependent GLUT1 and GLUT4 expression.


Assuntos
Diterpenos/toxicidade , Glucose/metabolismo , Cardiopatias/induzido quimicamente , Quinase I-kappa B/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , NF-kappa B/metabolismo , Fenantrenos/toxicidade , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/efeitos dos fármacos , Cardiotoxicidade , Linhagem Celular , Metabolismo Energético/efeitos dos fármacos , Compostos de Epóxi/toxicidade , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 4/genética , Transportador de Glucose Tipo 4/metabolismo , Cardiopatias/genética , Cardiopatias/metabolismo , Cardiopatias/patologia , Camundongos Knockout , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/deficiência , Proteína Supressora de Tumor p53/genética
2.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(3): 328-333, 2019 May.
Artigo em Chinês | MEDLINE | ID: mdl-31631598

RESUMO

Objective: To investigate the expression of miRNA-148b-3p and its target gene in the placenta between normal pregnant women and pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to explore the possible mechanism of glucose metabolism of offspring with maternal cholestasis. Methods: There were 30 cases of normal pregnant women and 30 cases of pregnant women with ICP recruited in the study, all of whom underwent cesarean delivery from Mar. 2017 to Jan. 2018. Placenta tissues, maternal blood and cord blood were collected in each case. Maternal blood and cord blood were sent for biochemical detection. miRNA of placenta tissues was extracted and qRT-PCR was used to measure the expression of miR-148b-3p in the placenta. Normal HTR-8 cells were transfected with miR-148b-3p inhibitor/mimics wrapped with lipofectaine3000. qRT-PCR was used to measure the expression of miR-148b-3p, and Western blot was used to measure the expression of glucose transporter 1 (GLUT1) after transfection. Results: Maternal fasting blood glucose (FPG) and its fetal cord blood insulin levels in the ICP group were significantly higher than those of control. The expression of miR-148b-3p in the placenta of ICP group was lower than that of control group ( P<0.05). Compared with inhibitor control group, the expression of miR-148b-3p was decreased in HTR-8 cells transfected with miR-148b-3p inhibitor ( P<0.05), while the expression of GLUT1 was increased ( P<0.05). Compared with mimics control group, the expression of miR-148b-3p was increased in HTR-8 cells transfected with miR-148b-3p mimics ( P<0.05), while the expression of GLUT1 was decreased ( P<0.05). Conclusion: miR-148b-3p might participate in glucose metabolism of offspring with maternal cholestasis through the negative regulation of GLUT1 expression in placental trophoblast cells.


Assuntos
Colestase Intra-Hepática/genética , Transportador de Glucose Tipo 1/genética , Glucose/metabolismo , MicroRNAs/genética , Placenta/citologia , Complicações na Gravidez/genética , Trofoblastos/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Gravidez
3.
Life Sci ; 232: 116588, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31226418

RESUMO

AIMS: Retinopathy is a neurodegenerative complication associating diabetes mellitus. Diabetic retinopathy (DR) is the primary reason of visual loss during early adulthood. DR has a complicated multifactorial pathophysiology initiated by hyperglycaemia-induced ischaemic neurodegenerative retinal changes, followed by vision-threatening consequences. The main therapeutic modalities for DR involve invasive delivery of intravitreal antiangiogenic agents as well as surgical interventions. The current work aimed to explore the potential anti-inflammatory and retinal neuroprotective effects of levetiracetam. MAIN METHODS: This study was performed on alloxan-induced diabetes in mice (n: 21). After 10 weeks, a group of diabetic animals (n: 7) was treated with levetiracetam (25 mg/kg) for six weeks. Retinal tissues were dissected and paraffin-fixed for examination using (1) morphometric analysis with haematoxylin and eosin (HE), (2) immunohistochemistry (GLUT1, GFAP and GAP43), and (3) RT-PCR-detected expression of retinal inflammatory and apoptotic mediators (TNF-α, IL6, iNOS, NF-κB and Tp53). KEY FINDINGS: Diabetic mice developed disorganized and debilitated retinal layers with upregulation of the gliosis marker GFAP and downregulation of the neuronal plasticity marker GAP43. Additionally, diabetic retinae showed increased transcription of NF-κB, TNF-α, IL6, iNOS and Tp53. Levetiracetam-treated mice showed downregulation of retinal GLUT1 with relief and regression of retinal inflammation and improved retinal structural organization. SIGNIFICANCE: Levetiracetam may represent a potential neuroprotective agent in DR. The data presented herein supported an anti-inflammatory role of levetiracetam. However, further clinical studies may be warranted to confirm the effectiveness and safety of levetiracetam in DR patients.


Assuntos
Retinopatia Diabética/tratamento farmacológico , Retinopatia Diabética/metabolismo , Proteína GAP-43/biossíntese , Transportador de Glucose Tipo 1/biossíntese , Levetiracetam/farmacologia , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Retinopatia Diabética/sangue , Retinopatia Diabética/genética , Modelos Animais de Doenças , Proteína GAP-43/genética , Transportador de Glucose Tipo 1/genética , Imuno-Histoquímica , Inflamação/metabolismo , Interleucina-6/metabolismo , Masculino , Camundongos , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Óxido Nítrico Sintase Tipo II/metabolismo , Retina/metabolismo , Doenças Retinianas/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
4.
Medicine (Baltimore) ; 98(18): e15428, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31045803

RESUMO

RATIONALE: The SLC2A1 gene encodes glucose transporter 1 on blood-brain barrier, which plays an important role in the energy supply for neurons. Mutations in SLC2A1 gene can cause many clinical syndromes, including glucose transporter type 1 deficiency syndrome and many types of epilepsy syndromes such as childhood absence epilepsy and myoclonic-atonic epilepsy, etc. Ketogenic diet has been proved to be very effective on those cases. Clinically, SLC2A1 gene mutations are quite rare. PATIENT CONCERNS: Repeated unconsciousness and bilateral limb weakness lasted for 3 years. DIAGNOSES: Myoclonic-atonic epilepsy. LESSONS: After taking whole exome sequencing, we found out that there is a de novo insertion mutation in the patient's SLC2A1 gene, leading to frameshift. As a result, ketogenic diet (2:1, 4 times a day) was used as the treatment. As for the patient, total calories intake per day was controlled at 1190 kcal. The calories per kg per day were 66.11 kcal/kg. The amount of ketone bodies was controlled at 2 to 3 mmol/L and the concentration of plasma glucose was controlled at 4 to 5 mmol/L. OUTCOMES: After the launch of ketogenic diet, the patient has been seizure free for nearly a year and stopped all his antiepileptic drugs. CONCLUSION: Our case suggests that gene examination is very important part of the diagnosis of epilepsy etiology and epilepsy syndromes. Ketogenic diet should be considered as the first line therapy with SLC2A1 gene mutations.


Assuntos
Dieta Cetogênica/métodos , Epilepsias Mioclônicas/dietoterapia , Epilepsias Mioclônicas/genética , Transportador de Glucose Tipo 1/genética , Pré-Escolar , Humanos , Masculino , Mutação
5.
Anticancer Res ; 39(5): 2351-2360, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31092427

RESUMO

BACKGROUND/AIM: Hepatic hemangiomas (HH) can show an aggressive course with significant complications. Prognostic markers that identify an aggressive course are entirely absent. Since we have showed that Hedgehog signaling is overexpressed in aggressive hemangiomas of the skin. Here, we hypothesize that it is also altered in aggressive HH. MATERIALS AND METHODS: Immunohistological staining for GLUT1 and quantitative PCR was performed in seven specimens with aggressive HH. For comparison, we included specimens of kaposiform hemangioendothelioma (KHE), skin hemangioma and normal liver tissue. RESULTS: Overexpression of the Hedgehog signaling components SHH and GLI2 and its target gene FOXA2 in HH were similar to those found in aggressive skin hemangioma and KHE, their expression being significantly higher than in mild skin hemangioma. High expression levels of SHH and FOXA2 positively correlated with HH, but not with normal liver tissue. CONCLUSION: Hedgehog signaling is up-regulated in aggressive HH. This finding may lead to a biomarker allowing early intervention.


Assuntos
Proteínas Hedgehog/genética , Hemangioma/genética , Fator 3-beta Nuclear de Hepatócito/genética , Neoplasias Hepáticas/genética , Proteínas Nucleares/genética , Proteína Gli2 com Dedos de Zinco/genética , Biomarcadores Tumorais/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Transportador de Glucose Tipo 1/genética , Hemangioendotelioma/genética , Hemangioendotelioma/patologia , Hemangioma/patologia , Humanos , Lactente , Recém-Nascido , Síndrome de Kasabach-Merritt/genética , Síndrome de Kasabach-Merritt/patologia , Fígado/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Sarcoma de Kaposi/genética , Sarcoma de Kaposi/patologia , Transdução de Sinais/genética
6.
J Exp Clin Cancer Res ; 38(1): 157, 2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30975171

RESUMO

BACKGROUND: Prostate cancer (PCa) remains a challenge worldwide. Due to the development of castration-resistance, traditional first-line androgen deprivation therapy (ADT) became powerlessness. Epidermal growth factor receptor (EGFR) is a well characterized therapeutic target to treat colorectal carcinoma and non-small cell lung cancer. Increasing studies have unraveled the significance of EGFR and its downstream signaling in the progression of castration-resistant PCa. METHOD: MTS, colony formation and Edu staining assays were used to analyze the cell proliferation of PCa cells. Flow cytometry was used to analyze PCa cell cycle distribution and cell apoptosis. Western blot was used to measure the expression of key proteins associated with cell cycle progression, apoptosis and EGFR signaling pathways. Transfection of exogenous small interfering RNA (siRNA) or plasmid was used to intervene specific gene expression. Nude mouse model was employed to test the in vivo effect of Spautin-1. RESULTS: The current study reveals that Spautin-1, a known inhibitor of ubiquitin-specific peptidase 10 (USP10) and USP13, inhibits EGFR phosphorylation and the activation of its downstream signaling. Inhibition of EGFR signaling induced by Spautin-1 leads to cell cycle arrest and apoptosis of PCa in a USP10/USP13 independent manner. The application of Spautin-1 reduces the expression of glucose transporter 1 (Glut1) and dramatically induces cell death under glucose deprivation condition. In vivo experiments show a potent anti-tumor effect of Spautin-1 alone and in combination with Enzalutamide. CONCLUSION: This study demonstrates the therapeutic potential of EGFR signaling inhibition by the use of Spautin-1 for PCa treatment.


Assuntos
Benzilaminas/farmacologia , Neoplasias da Próstata/metabolismo , Quinazolinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Biomarcadores Tumorais , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclina D1/genética , Modelos Animais de Doenças , Receptores ErbB/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glucose/metabolismo , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Masculino , Camundongos , Modelos Biológicos , Terapia de Alvo Molecular , Feniltioidantoína/análogos & derivados , Feniltioidantoína/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/patologia , Ensaios Antitumorais Modelo de Xenoenxerto
7.
Nat Commun ; 10(1): 1915, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-31015424

RESUMO

Bromodomains (BRDs) are conserved protein interaction modules which recognize (read) acetyl-lysine modifications, however their role(s) in regulating cellular states and their potential as targets for the development of targeted treatment strategies is poorly understood. Here we present a set of 25 chemical probes, selective small molecule inhibitors, covering 29 human bromodomain targets. We comprehensively evaluate the selectivity of this probe-set using BROMOscan and demonstrate the utility of the set identifying roles of BRDs in cellular processes and potential translational applications. For instance, we discovered crosstalk between histone acetylation and the glycolytic pathway resulting in a vulnerability of breast cancer cell lines under conditions of glucose deprivation or GLUT1 inhibition to inhibition of BRPF2/3 BRDs. This chemical probe-set will serve as a resource for future applications in the discovery of new physiological roles of bromodomain proteins in normal and disease states, and as a toolset for bromodomain target validation.


Assuntos
Antineoplásicos/farmacologia , Células Epiteliais/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/farmacologia , Acetilação , Sequência de Aminoácidos , Antineoplásicos/química , Linhagem Celular Tumoral , Epigênese Genética , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Glucose/deficiência , Transportador de Glucose Tipo 1/antagonistas & inibidores , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Glicólise/efeitos dos fármacos , Glicólise/genética , Ensaios de Triagem em Larga Escala , Histonas/genética , Histonas/metabolismo , Humanos , Glândulas Mamárias Humanas/metabolismo , Glândulas Mamárias Humanas/patologia , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Transdução de Sinais , Bibliotecas de Moléculas Pequenas/química , Relação Estrutura-Atividade
8.
Toxicol Lett ; 310: 23-30, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30980912

RESUMO

The aim of this study was to determine whether Pb affects glucose metabolism in the hippocampus of rats. Male Sprague-Dawley rats aged 21 days were orally administered a 0.1%, 0.2%, or 0.3% lead acetate solution in deionized water for 65 days. Then, the weight of the rats; brain Pb content; brain glucose levels; activities of hexokinase, fructose-6-phosphate kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase; expression of genes related to the insulin signaling pathway; as well as the gene and protein expression of glucose transporter (GLUT)-1 and GLUT-3 in the hippocampus were evaluated. The results showed that Pb content in the brain tissue of rats in the dose groups significantly increased, whereas the body weight gain, activities of glucose metabolism-related enzymes, and expression of the insulin signaling pathway-related genes significantly decreased compared to the corresponding values in the control group. In comparison with the control group, the brain glucose levels increased significantly in the low-dose group, but there were no significant differences with the middle- and high-dose groups. Furthermore, the mRNA of GLUT-1 in the three dose groups and the GLUT-3 in the middle- and high-dose groups rose markedly, while the GLUT-1 and GLUT-3 protein expression significantly increased in the middle- and high-dose groups and in the high-dose group, respectively. Taken together, the results showed that Pb exposure resulted in a lower body weight gain, higher brain Pb content and also affected brain glucose metabolism and the insulin signaling pathway.


Assuntos
Metabolismo Energético/efeitos dos fármacos , Glucose/metabolismo , Hipocampo/efeitos dos fármacos , Insulina/metabolismo , Compostos Organometálicos/toxicidade , Transdução de Sinais/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 3/genética , Transportador de Glucose Tipo 3/metabolismo , Glucosefosfato Desidrogenase/genética , Glucosefosfato Desidrogenase/metabolismo , Hexoquinase/genética , Hexoquinase/metabolismo , Hipocampo/metabolismo , Hipocampo/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Fosfofrutoquinase-2/genética , Fosfofrutoquinase-2/metabolismo , Piruvato Quinase/genética , Piruvato Quinase/metabolismo , Ratos Sprague-Dawley , Medição de Risco , Ganho de Peso/efeitos dos fármacos
9.
Kaohsiung J Med Sci ; 35(1): 24-32, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30844141

RESUMO

Several studies have reported the beneficial effects of Lawsonia inermis on wound healing, but the mechanism of action is still unknown. This study aimed to investigate the effectiveness of a new ointment formulation of hydroethanolic extract leaves of L. inermis on wound healing by gene expression of glucose transporter-1 (Glut-1) and insulin-like growth factor I (Igf-1) in Wistar rats. The animals were topically treated with different doses of L. inermis. An experimentally induced circular excisional wound model of 314 mm2 surface area was surgically created. The percentage of wound contraction and histopathological changes was assessed at different time points following wound induction. The expression of Glut-1 and Igf-1 was evaluated by reverse-transcription PCR. Topical administration of L. inermis, dose dependently, shortened inflammatory phase, accelerated cellular proliferation, and enhanced wound contraction ratio. It also improved revascularization, collagen deposition, and re-epithelialization rate and promoted intracytoplasmic carbohydrate storage (P < 0.05). Moreover, the mRNA levels of Igf-1 and Glut-1 were significantly higher in the L. inermis-treated groups than the control group (P < 0.05). Topical administration of L. inermis promoted the healing process by reducing tissue inflammation and increasing glucose uptake, which was mediated by up-regulating the expression of Igf-1 and Glut-1.


Assuntos
Etanol/química , Glucose/metabolismo , Inflamação/patologia , Lawsonia (Planta)/química , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Água/química , Cicatrização/efeitos dos fármacos , Administração Tópica , Fosfatase Alcalina/metabolismo , Antioxidantes/metabolismo , Carboidratos/análise , Epiderme/efeitos dos fármacos , Epiderme/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Flavonoides/análise , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Fenóis/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reepitelização/efeitos dos fármacos
10.
Hum Cell ; 32(2): 193-201, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30868406

RESUMO

Bladder cancer (BC) is one of the most common tumors. Metabolic reprogramming is a feature of neoplasia and tumor growth. Understanding the metabolic alterations in bladder cancer may provide new directions for bladder cancer treatment. Sirtuin 1 (SIRT1) is a lysine deacetylase of multiple targets including metabolic regulators. In pancreatic cancer, the loss of SIRT1 is accompanied by a decreased expression of proteins in the glycolysis pathway, such as GLUT1, and cancer cell proliferation. Thus, we hypothesize that SIRT1 may interact with GLUT1 to modulate the proliferation and glycolysis phenotype in bladder cancer. In the present study, the expression of SIRT1 and GLUT1 was upregulated in BC tissues and cell lines and positively correlated in tissue samples. SIRT1 overexpression or GLUT1 overexpression alone was sufficient to promote cell proliferation and glucose uptake in BC cells. EX527, a specific inhibitor of SIRT1, exerted an opposing effect on bladder cancer proliferation and glucose uptake. The effect of EX527 could be partially reversed by GLUT1 overexpression. More importantly, SIRT1 overexpression significantly promoted the transcriptional activity and expression of GLUT1, indicating that SIRT1 increases the transcription activity and expression of GLUT1, therefore, promoting the cell proliferation and glycolysis in BC cells. Our study first reported that SIRT1/GLUT1 axis promotes bladder cancer progression via regulation of glucose uptake.


Assuntos
Expressão Gênica/genética , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Glucose/metabolismo , Sirtuína 1/genética , Sirtuína 1/fisiologia , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/patologia , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Glicólise/genética , Glicólise/fisiologia , Humanos , Regulação para Cima , Neoplasias da Bexiga Urinária/metabolismo
11.
Cancer Sci ; 110(5): 1705-1714, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30861255

RESUMO

Glucose transporter 1 (GLUT1) expression is a prognostic marker for esophageal squamous cell carcinoma (ESCC). Recent work on GLUT1 and development of specific inhibitors supports the feasibility of GLUT1 inhibition as a treatment for various cancers. The anti-proliferative effects of GLUT1-specific small interfering RNA (siRNA) and a GLUT1 inhibitor were evaluated in ESCC cell lines. Expression of pro-proliferative and anti-proliferative signaling and effector molecules was examined by western blotting and quantitative RT-PCR. GLUT1 expression in pretreatment clinical biopsy samples was measured by immunohistochemistry and correlated with various clinicopathological parameters and response to chemotherapy. The reduction in standardized uptake value (SUV) of 18 F-fluoro-deoxyglucose was calculated using the formula: ([pretreatment SUVmax  - posttreatment SUVmax ]/pretreatment SUVmax ) × 100. GLUT1-specific siRNA expression in ESCC cells inhibited their proliferation, increased expression of p27kip, and decreased expression of cyclin-dependent kinase 6, pyruvate kinase muscle isozyme M2, lactate dehydrogenase A and phospho-ERK1/2. Suppression of GLUT1 by siRNA increased low-dose cisplatin-induced inhibition of proliferation of TE-11 ESCC cells, which express high GLUT1 levels. Similarly, BAY-876, a GLUT1 inhibitor, enhanced cisplatin-mediated inhibition of ESCC cell proliferation. GLUT1 expression in pretreatment biopsy samples was associated with the response to chemotherapy as well as the pathological tumor stage and histological response grade after esophagectomy. Finally, GLUT1-negative tumors showed a significantly larger reduction in SUVmax (61.2% ± 4.5%) compared with GLUT1-positive tumors (46.2% ± 4.4%). GLUT1 expression may be a surrogate marker of response to chemotherapy, and inhibition of GLUT1 may be a potential novel therapy for ESCC patients.


Assuntos
Resistencia a Medicamentos Antineoplásicos , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago/metabolismo , Transportador de Glucose Tipo 1/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/cirurgia , Carcinoma de Células Escamosas do Esôfago/tratamento farmacológico , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/cirurgia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Transportador de Glucose Tipo 1/genética , Humanos , Masculino , Pirazóis/farmacologia , Quinolinas/farmacologia , RNA Interferente Pequeno/farmacologia , Transdução de Sinais/efeitos dos fármacos
12.
J Biosci ; 44(1)2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30837370

RESUMO

Among the last consequences of metabolic syndrome are cardiovascular complications such as infarcts. The hypoxic heart switches its lipid-based metabolism to carbohydrates, and a glucose-insulin-potassium (GIK) solution can be the metabolic support to protect the organ. Due to the physiology and cardiac risks associated with the metabolic syndrome, we studied the effect of GIK solution during hypoxia in a metabolic syndrome model by observing the participation of glucose transporters (GLUTs). The metabolic syndrome characteristics were established by giving a 30% sucrose drinking solution to Wistar rats for 24 weeks. The GIK solution's effect on myocyte glucose uptake during hypoxia and oxygenation was observed using a colorimetric method, and Western blot technique visualized the GLUT participation. Oxygenated control myocytes consumed 1.7 +/- 0.2 µg of glucose per gram of fresh tissue per hour using the GLUT1, and during hypoxia, they incorporated 41.1% more glucose by GLUT1 and GLUT4. The GIK solution improved glucose uptake in oxygenation by 70.5% through GLUT1. In hypoxia, the uptake was 21% more than the hypoxic control group and by both GLUTs too. Oxygenated metabolic syndrome myocytes uptake was similar to control cells but achieved by both carriers in oxygenation and hypoxia. Also, the GIK solution had a better response in both oxygenation (113%) and hypoxia (71%). Despite the metabolic energy disorders of this syndrome, the GIK solution protects cardiomyocytes, in conditions of hypoxia, through the modulation of both GLUTs. So, this solution can be considered a useful resource during a heart attack in cases of metabolic syndrome.


Assuntos
Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 4/genética , Síndrome Metabólica/prevenção & controle , Infarto do Miocárdio/prevenção & controle , Animais , Hipóxia Celular/efeitos dos fármacos , Modelos Animais de Doenças , Metabolismo Energético/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Glucose/administração & dosagem , Glucose/metabolismo , Humanos , Insulina/administração & dosagem , Síndrome Metabólica/complicações , Síndrome Metabólica/genética , Síndrome Metabólica/metabolismo , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Potássio/administração & dosagem , Ratos
13.
Mol Biol Rep ; 46(2): 1727-1736, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30725350

RESUMO

Metabolic syndrome is an agglomeration of disorders including obesity, diabetes and cardiovascular diseases and characterized as chronic mild inflammation which elevates the circulatory inflammatory markers. This could be due to mitochondrial dysfunction, oxidative stress and hypoxia as a consequence of high fat diet (HFD) intake. The present study focuses on the effects of HFD on lactate and mitochondrial metabolism as well as tissue dependent changes in glucose transporter (GLUT) expression in liver, skeletal muscles and adipose tissue of mouse. Lactate dehydrogenase (LDH) and mitochondrial dysfunction established the link between the occurrences of metabolic stress due to HFD. In this work, it was observed that chronic HFD administration aggravated the metabolic alterations by causing reduced ATP production, imbalanced oxidative stress and altered class 1 GLUTs expression. Chronic HFD significantly reduced (p < 0.001) the superoxide dismutase (SOD), catalase (CAT) activities alongside elevated liver injury markers AST and ALT. This in turn causes decreased ATP/ADP ratio, mitochondrial dysfunction and exacerbated LDH levels. This imbalance further led to altered GLUT expression in hepatic cells, adipose tissue and skeletal muscles. HFD significantly (p < 0.001) upregulated the GLUT 1 and 3 expressions while significant downregulated (p < 0.001) GLUT 2 and 4 expression in liver, skeletal muscles and white adipose tissue. These results revealed the link between class 1 GLUTs, mitochondrial dysfunction and HFD-induced metabolic disorder. It can be concluded that HFD impacts mitochondrial metabolism and reprograms tissue-dependent glucose transporter.


Assuntos
Transportador de Glucose Tipo 1/biossíntese , Transportador de Glucose Tipo 1/genética , Mitocôndrias/metabolismo , Tecido Adiposo/metabolismo , Animais , Peso Corporal , Dieta Hiperlipídica , Regulação da Expressão Gênica , Transportador de Glucose Tipo 1/metabolismo , Hepatócitos/metabolismo , Inflamação/metabolismo , Resistência à Insulina/fisiologia , Ácido Láctico/metabolismo , Fígado/metabolismo , Masculino , Síndrome Metabólica/genética , Síndrome Metabólica/metabolismo , Camundongos , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Estresse Oxidativo
14.
EBioMedicine ; 41: 370-383, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30799198

RESUMO

BACKGROUND: Cancer-associated fibroblasts (CAFs) are the predominant residents in the breast tumor microenvironment. In our work, we found activation of DNA damage-independent ATM (oxidized ATM), enhanced glycolysis and aberrant metabolism-associated gene expressions in breast CAFs. Nevertheless, whether and how oxidized ATM regulates the glycolytic activity of CAFs keep in unveil. Recently, a reverse Warburg effect was observed in tumor tissues, in which host cells (such as CAFs, PSCs) in the tumor microenvironment have been found to "fuel" the cancer cells via metabolites transfer. However, the molecular mechanisms of the metabolites from stromal cells playing a role to the progression of cancer cells remain to be determined. METHODS: Oxidized ATM activation in stromal CAFs was assessed by western blotting and immunofluorescence. The increased glycolytic ability of CAFs was validated by measurements of OCR and ECAR and detections of glucose consumption and lactate production. Kinase assay and western blotting were performed to confirm the phosphorylation of GLUT1. The membrane location of phosphorylated GLUT1 was determined by biotin pull-down assay and immunofluorescence staining. The regulation of PKM2 through oxidized ATM was evaluated by western blots. In addition, the impact of lactate derived from hypoxic CAFs on cancer cell invasion was investigated both in vitro (transwell assays, western blots) and in vivo (orthotopic xenografts). FINDINGS: Hypoxia-induced oxidized ATM promotes glycolytic activity of CAFs by phosphorylating GLUT1 at S490 and increasing PKM2 expression. Moreover, lactate derived from hypoxic CAFs, acting as a metabolic coupling between CAFs and breast cancer cells, promotes breast cancer cell invasion by activating the TGFß1/p38 MAPK/MMP2/9 signaling axis and fueling the mitochondrial activity in cancer cells. INTERPRETATION: Our work shows that oxidized ATM-mediated glycolysis enhancement in hypoxic stromal fibroblasts plays an essential role in cancer cell invasion and metastasis and may implicate oxidized ATM as a target for breast tumor treatment. FUND: This research was supported by National Natural Science Foundation of China.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Ácido Láctico/metabolismo , Animais , Proteínas Mutadas de Ataxia Telangiectasia/antagonistas & inibidores , Proteínas Mutadas de Ataxia Telangiectasia/genética , Neoplasias da Mama/patologia , Fibroblastos Associados a Câncer/citologia , Fibroblastos Associados a Câncer/metabolismo , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Hipóxia Celular , Movimento Celular , Células Cultivadas , Feminino , Glucose/metabolismo , Transportador de Glucose Tipo 1/antagonistas & inibidores , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Glicólise , Humanos , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Mitocôndrias/metabolismo , Fosforilação Oxidativa , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Hormônios Tireóideos/genética , Hormônios Tireóideos/metabolismo
15.
Int J Biol Macromol ; 129: 601-607, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30738168

RESUMO

After spinal cord injury, microglial cells are activated and converted to an M1 phenotype. Emerging evidence supports the hypothesis that glucose reprogramming accompanies microglial activation. What contributes to the activation of microglia and glucose reprogramming, however, remains unclear. In the current study, we investigated the role and underlying mechanism of a-synuclein in regulating the aerobic glycolysis in microglia. We found that a-synuclein contributed to the reprogramming of glucose metabolism in microglia by promoting glycolysis and inhibiting mitochondrial biogenesis and oxidative phosphorylation. Further studies demonstrated that pyruvate kinase M2 (PKM2), a rate-limiting enzyme in glycolysis, mediated glucose reprogramming regulated by a-synuclein. A co-immunoprecipitation assay and Western blot assay demonstrated that a-synuclein interacted with PKM2. Further studies demonstrated that knockdown of PKM2 in a-synuclein-exposed microglia markedly reduced glycolysis and lactate production. Additionally, a-synuclein exposure promoted migration abilities in glucose-cultured microglia, whereas migration ability was suppressed in PKM2 knockdown microglia. Additionally, the PKM2 activator TEPP-46 promoted migration ability in a-synuclein-treated microglia, compared to treatment with a-synuclein alone. In conclusion, we demonstrate a PKM2-dependent glycolysis of a-synuclein in microglial.


Assuntos
Movimento Celular/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Microglia/citologia , Microglia/efeitos dos fármacos , Piruvato Quinase/metabolismo , alfa-Sinucleína/farmacologia , Animais , Glucose/metabolismo , Transportador de Glucose Tipo 1/genética , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/genética , Humanos , Microglia/metabolismo , Piruvato Quinase/deficiência , Piruvato Quinase/genética , Ratos , Ratos Sprague-Dawley , Regulação para Cima/efeitos dos fármacos
16.
Diabetes ; 68(5): 932-938, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30765335

RESUMO

Patients with type 1 diabetes mellitus (T1DM) have increased thrombosis and platelet activation. The mechanisms for platelet hyperactivation in diabetes are incompletely understood. T1DM is accompanied by hyperglycemia, dyslipidemia, and increased inflammation in addition to an altered hormonal milieu. In vitro analysis of platelets revealed that normal glucose reduces platelet activation whereas hyperglycemic conditions increase platelet activation. We therefore hypothesized that hyperglycemia increases platelet glucose utilization, which increases platelet activation to promote thrombosis. Glucose uptake and glycolysis were increased in platelets isolated from mice given streptozotocin (STZ) to induce T1DM in concert with induction of GLUT3. Platelets from STZ-induced diabetic mice exhibited increased activation after administration of protease-activated receptor 4 peptide and convulxin. In contrast, platelets isolated from GLUT1 and GLUT3 double-knockout (DKO) mice, which lack the ability to use glucose, failed to increase activation in hyperglycemic mice. Diabetic mice displayed decreased survival in a collagen/epinephrine-induced pulmonary embolism model of in vivo platelet activation relative to nondiabetic controls. Survival after pulmonary embolism was increased in diabetic DKO mice relative to nondiabetic controls. These data reveal that increased platelet glucose metabolism in vivo contributes to increased platelet activation and thrombosis in a model of T1DM.


Assuntos
Plaquetas/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Glucose/metabolismo , Animais , Diabetes Mellitus Tipo 1/genética , Modelos Animais de Doenças , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 3/genética , Transportador de Glucose Tipo 3/metabolismo , Masculino , Camundongos , Camundongos Knockout , Ativação Plaquetária/genética , Ativação Plaquetária/fisiologia , Embolia Pulmonar/metabolismo , Embolia Pulmonar/mortalidade
17.
Eur J Paediatr Neurol ; 23(2): 329-332, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30616884

RESUMO

SLC2A1 mutations cause glucose transporter type 1 deficiency syndrome, whose phenotypic spectrum is a continuum, ranging from classic to variant phenotypes, the latter accounting for about 10% of cases. Very few SLC2A1-mutated patients with a spastic paraplegia phenotype have been reported so far, and they are associated with paroxysmal choreo-athetosis (i.e., DYT9). The authors describe two sporadic children with pure and complex hereditary spastic paraplegia (HSP) without paroxysmal non-epileptic movement disorders harboring heterozygous de novo SLC2A1 pathogenic variants. These patients have been identified by a targeted panel for HSP among 140 pediatric- and adult-onset unrelated cases with pure and complex HSP, thus indicating an overall prevalence of 1.4% of SLC2A1 mutations, which increases to 3% if only pediatric-onset patients are considered. The implications of these findings in the diagnostic work-up of HSP patients are discussed.


Assuntos
Transportador de Glucose Tipo 1/genética , Paraplegia Espástica Hereditária/genética , Erros Inatos do Metabolismo dos Carboidratos/genética , Criança , Feminino , Humanos , Masculino , Proteínas de Transporte de Monossacarídeos/deficiência , Proteínas de Transporte de Monossacarídeos/genética , Mutação , Linhagem , Fenótipo
18.
J Biol Chem ; 294(10): 3760-3771, 2019 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-30617181

RESUMO

Most clear cell renal cell carcinomas (ccRCCs) have inactivation of the von Hippel-Lindau tumor suppressor protein (pVHL), resulting in the accumulation of hypoxia-inducible factor α-subunits (HIF-α) and their downstream targets. HIF-2α expression is particularly high in ccRCC and is associated with increased ccRCC growth and aggressiveness. In the canonical HIF signaling pathway, HIF-prolyl hydroxylase 3 (PHD3) suppresses HIF-2α protein by post-translational hydroxylation under sufficient oxygen availability. Here, using immunoblotting and immunofluorescence staining, qRT-PCR, and siRNA-mediated gene silencing, we show that unlike in the canonical pathway, PHD3 silencing in ccRCC cells leads to down-regulation of HIF-2α protein and mRNA. Depletion of other PHD family members had no effect on HIF-2α expression, and PHD3 knockdown in non-RCC cells resulted in the expected increase in HIF-2α protein expression. Accordingly, PHD3 knockdown decreased HIF-2α target gene expression in ccRCC cells and expression was restored upon forced HIF-2α expression. The effect of PHD3 depletion was pinpointed to HIF2A mRNA stability. In line with these in vitro results, a strong positive correlation of PHD3 and HIF2A mRNA expression in ccRCC tumors was detected. Our results suggest that in contrast to the known negative regulation of HIF-2α in most cell types, high PHD3 expression in ccRCC cells maintains elevated HIF-2α expression and that of its target genes, which may enhance kidney cancer aggressiveness.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Carcinoma de Células Renais/patologia , Prolina Dioxigenases do Fator Induzível por Hipóxia/metabolismo , Neoplasias Renais/patologia , Linhagem Celular Tumoral , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Inativação Gênica , Transportador de Glucose Tipo 1/genética , Humanos , Prolina Dioxigenases do Fator Induzível por Hipóxia/deficiência , Prolina Dioxigenases do Fator Induzível por Hipóxia/genética , Estabilidade Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
19.
Epilepsy Res ; 150: 70-77, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30660939

RESUMO

Epilepsy with myoclonic-atonic seizures (EMAS) accounts for 1-2% of all childhood-onset epilepsies. EMAS has been shown to have an underlying genetic component, however the genetics of this disorder is not yet well understood. The purpose of this study was to review genetic testing results for a cohort of EMAS patients. A retrospective chart review was conducted for 77 patients evaluated at Children's Hospital Colorado with a potential diagnosis of EMAS. Genetic testing and biochemical testing was reviewed. Family history data was also collected. Seventy-seven percent of the cohort had at least one genetic test performed, and a molecular diagnosis was reached for six patients. Thirty-seven patients had a microarray, six of which identified a copy number variant. Only one was felt to contribute to the phenotype (2p16.3 deletion including NRXN1). Fifty-one patients had an epilepsy panel, two of which were positive (likely pathogenic variant in SCN1A, pathogenic variant in GABRG2). Of the six patients who had whole exome sequencing, two were negative, three were positive or likely positive, and one had multiple variants not felt to explain the phenotype. While EMAS is widely accepted to have a strong genetic component, the diagnostic yield of genetic testing remains low. This may be because several genes now thought to be associated with EMAS are not included on the more commonly ordered epilepsy panels, or have only recently been added to them.


Assuntos
Epilepsia/complicações , Epilepsia/genética , Convulsões/complicações , Convulsões/genética , Moléculas de Adesão Celular Neuronais/genética , Estudos de Coortes , Polimerase do DNA Mitocondrial/genética , Proteínas de Ligação a DNA/genética , Eletroencefalografia , Saúde da Família , Feminino , Testes Genéticos , Transportador de Glucose Tipo 1/genética , Humanos , Masculino , Mutação/genética , Canal de Sódio Disparado por Voltagem NAV1.1/genética , Proteínas do Tecido Nervoso/genética , Receptores de GABA-A/genética , Sequenciamento Completo do Exoma
20.
Phytomedicine ; 54: 120-131, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30668361

RESUMO

BACKGROUND: Emerging hallmark of cancer is reprogrammed cellular metabolism, increased glycolytic metabolism is physiological characteristic of human malignant neoplasms. Saponin monomer 13 of the dwarf lilyturf tuber (DT-13) is the main steroidal saponin from Liriopes Radix, which has been reported to exert anti-inflammation and anti-tumor activities but low toxicity to normal tissue. However, the effect of DT-13 on metabolism process is still unclear. PURPOSE: This study aims to characterize the role of DT-13 in glucose metabolism in colorectal cancer cells, and investigate whether the metabolism process is involved in the anti-cancer response of DT-13. METHODS: Colony formation assay was employed to determine anti-proliferative effect induced by DT-13 at 2.5, 5, 10 µM. Apoptosis and cell cycle arrest were detected by Annexin V/PI staining and PI staining, respectively. Genetic inhibition of glycolytic metabolism was carried out by knockdown of GLUT1. Orthotopic implantation mouse model of colorectal cancer was used to assess in vivo antitumor effect of DT-13 (0.625, 1.25, 2.5 mg/kg). The chemoprevention effect of DT-13 (10mg/kg) was evaluated by using C57BL/6J APCmin mice model. Glycolytic-related key enzymes and AMPK pathway were detected by using quantitative real-time PCR, western blotting, and immunohistochemical staining. RESULTS: Our results showed that cell proliferation was significantly inhibited by DT-13 in a dose-dependent manner. DT-13 inhibited glucose uptake, ATP generation, and reduced lactate production. Furthermore, DT-13 remarkably inhibited GLUT1 expression in both mRNA and protein levels. Knocking down of GLUT1 led to reduced inhibition of glucose uptake after DT-13 treatment. Moreover, deletion of GLUT1 decreased inhibitory ratio of DT-13 on cancer growth. Orthotopic implantation mouse model of colorectal cancer further confirmed that DT-13 inhibited colorectal cancer growth via blocking GLUT1 in vivo. In addition, C57BL/6J APCmin mice model revealed that DT-13 dramatically reduced the total number of spontaneous adenomas in intestinal, which further confirmed the anti-tumor activity of DT-13 in colorectal cancer. Furthermore, the mechanistically investigation showed DT-13 activated AMPK and inhibited m-TOR to block cancer growth in vitro. CONCLUSION: DT-13 is a potent anticancer agent for colorectal cancer.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Saponinas/farmacologia , Serina-Treonina Quinases TOR/metabolismo , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Transportador de Glucose Tipo 1/antagonistas & inibidores , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Glicólise , Humanos , Liriope (Planta)/química , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
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