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1.
Adv Exp Med Biol ; 1217: 363-372, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31898238

RESUMO

MLN4924, also known as pevonedistat, is a highly selective small-molecule inhibitor of NEDD8 (neuronal precursor cell-expressed developmentally downregulated protein 8)-activating enzyme (NAE) to block the entire neddylation modification cascade, leading to inactivation of cullin-RING ligases (CRLs), since activation of CRLs requires cullin neddylation. MLN4924 showed impressive anticancer activity in many preclinical studies and is currently in several Phase I/II clinical trials for anticancer therapy as a single agent or in combination with chemotherapeutic drugs.In addition to well-characterized anti-neddylation activity, recent studies showed that MLN4924 has several neddylation-independent activities. First, MLN4924 triggers EGFR dimerization to activate EGFR and its downstream RAS/MAPK and PI3K/AKT1 signals, leading to enhanced tumor sphere formation, accelerated EGF-mediated wound healing, and inhibited ciliogenesis. Second, MLN4924 induces PKM2 tetramerization to promote glycolysis, thus affecting energy metabolism. Third, MLN4924 inhibits the interaction between ACT1 (NF-κB activator 1) and TRAF6 (tumor necrosis factor receptor-associated factor 6) and attenuates IL-17A-mediated activation of NF-κB to reduce pulmonary inflammation. Fourth, MLN4924 inhibits IRF3 binding to the IFN-ß promoter to inhibit IFN-ß production. And finally, MLN4924 activates the JNK signaling pathway to reduce c-FLIP levels, thus enhancing TRAIL-induced apoptosis. This chapter will summarize these neddylation-independent activities of MLN4924 and discuss the underlying mechanisms and potential therapeutic applications.


Assuntos
Ciclopentanos/farmacologia , Ciclopentanos/uso terapêutico , Pirimidinas/farmacologia , Pirimidinas/uso terapêutico , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Proteínas de Transporte/metabolismo , Receptores ErbB/metabolismo , Glicólise/efeitos dos fármacos , Humanos , Interferon beta/biossíntese , Proteínas de Membrana/metabolismo , Proteína NEDD8/metabolismo , Pneumonia/tratamento farmacológico , Hormônios Tireóideos/metabolismo
2.
Toxicol Lett ; 323: 19-24, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31962156

RESUMO

Cultured kidney cells maintained in conventional growth media with high glucose levels exhibit increased glycolytic activity compared to the cells in vivo. In contrast, renal proximal tubules utilize substrates such as ketone bodies and rely on mitochondrial oxidative phosphorylation. LLC-PK1 cells maintain many features of the proximal tubule but are exposed to glucose concentrations ranging from 17 to 25 mM. This may impact their reliability in predicting mitochondrial toxicity. This study is designed to test the impact of the ketone body acetoacetate on metabolic characteristics of LLC-PK1 cells. Basal respiration, maximal respiration, spare respiratory capacity and ATP-linked respiration were significantly increased in cells grown in growth medium supplemented with 5 mM acetoacetate. In contrast, glycolytic capacity, as well as glycolytic reserve were significantly reduced in the acetoacetate group. There was an increased expression in biomarkers of mitochondrial biogenesis, and an increase in mitochondrial protein expression. Cells grown in medium complemented with acetoacetate displayed a significantly lower LC50 when treated with clotrimazole and diclofenac. There was a marked increase in uncoupled respiration in the presence of diclofenac, while clotrimazole and ciprofibrate significantly decreased respiration in the acetoacetate. The results indicate that acetoacetate complemented media can alter cellular metabolism and increase sensitization to toxicants.


Assuntos
Acetoacetatos/farmacologia , Rim/efeitos dos fármacos , Animais , Células Cultivadas , Clotrimazol/toxicidade , Diclofenaco/toxicidade , Ácidos Fíbricos/toxicidade , Glicólise/efeitos dos fármacos , Rim/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Oxirredução , Suínos
3.
Toxicol Lett ; 318: 74-85, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31654802

RESUMO

Metabolic flexibility defines the capacity of cells to respond to changes in nutrient status. Mitochondria are important mediators of metabolic flexibility and dysfunction is associated with metabolic inflexibility and pathology. Foodborne toxins are often overlooked as potential factors contributing to metabolic toxicity. Fusaric acid (FA), a neglected mycotoxin, is known to disrupt mitochondrial function. The aim of this study was to investigate the molecular mechanisms underlying a metabolic switch in response to FA. This study investigated the effects of FA on energy homeostasis in cultured human liver (HepG2) cells. HepG2 cells poised to undergo oxidative and glycolytic metabolism were exposed to a range of FA concentrations (4, 63 and 250 µg/mL) for 6 h. We determined mitochondrial toxicity, acetyl CoA levels and cell viability using luminometric, fluorometric and spectrophotometric methods. Expression of metabolic proteins (PDK1, PKM2, phosphorylated-PDH E1α and HIF-1α) and mRNAs (HIF-1α, PKM2, LDHa and PDK1) were determined using western blot and qPCR respectively. Our data connects a constitutive expression of HIF-1α in response to FA, to the inhibition of pyruvate decarboxylation through up-regulation of PDK-1 and phosphorylation of Pyruvate Dehydrogenase E1α subunit. Moreover, we highlight the potential of FA to induce a glucose "addiction" and phenotype reminiscent of the Warburg effect. The findings provide novel insights into the impact of this neglected foodborne mycotoxin in the dysregulation of energy metabolism.


Assuntos
Plasticidade Celular/efeitos dos fármacos , Microbiologia de Alimentos , Ácido Fusárico/toxicidade , Glicólise/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Células Hep G2 , Hepatócitos/metabolismo , Hepatócitos/patologia , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Hepáticas/patologia , Fenótipo , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Piruvato Desidrogenase (Lipoamida)/metabolismo
4.
Food Chem ; 305: 125439, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31499287

RESUMO

Compared to the control longans, hydrogen peroxide (H2O2)-treated longans exhibited higher index of pulp breakdown, higher fruit respiration rate, higher activities of pulp phosphohexose isomerase (PGI), succinate dehydrogenase (SDH), cytochrome C oxidase (CCO), ascorbic acid oxidase (AAO) and polyphenol oxidase (PPO), but lower activity of pulp nicotinamide adenine dinucleotide kinase (NADK). H2O2-treated longans also exhibited lower total activities of pulp glucose-6-phosphate dehydrogenase (G-6-PDH) and 6-phosphogluconate dehydrogenase (6-PGDH), lower levels of pulp NADP(H), but higher levels of pulp NAD(H). These data indicated that H2O2-stimulated longan pulp breakdown was owing to a decreased proportion of pentose phosphate pathway (PPP), the increased proportions of Embden-Meyerhof-Parnas pathway (EMP), tricarboxylic acid (TCA) cycle and cytochrome pathway (CCP) in total respiratory pathways. These findings further revealed that H2O2 could enhance respiration rate, and thus accelerate pulp breakdown occurrence and shorten the shelf life of longan fruit.


Assuntos
Peróxido de Hidrogênio/farmacologia , Sapindaceae/efeitos dos fármacos , Aldeído Oxidase/metabolismo , Ciclo do Ácido Cítrico/efeitos dos fármacos , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Armazenamento de Alimentos , Frutas/metabolismo , Glucosefosfato Desidrogenase/metabolismo , Glicólise/efeitos dos fármacos , NAD/metabolismo , Via de Pentose Fosfato/efeitos dos fármacos , Sapindaceae/metabolismo
5.
Braz J Med Biol Res ; 53(1): e8389, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31859908

RESUMO

Photodynamic therapy (PDT) promotes cell death, and it has been successfully employed as a treatment resource for neuropathic complications of diabetes mellitus (T1DM) and hepatocellular carcinoma. The liver is the major organ involved in the regulation of energy homeostasis, and in pathological conditions such as T1DM, changes in liver metabolic pathways result in hyperglycemia, which is associated with multiple organic dysfunctions. In this context, it has been suggested that chlorophyll-a and its derivatives have anti-diabetic actions, such as reducing hyperglycemia, hyperinsulinemia, and hypertriglyceridemia, but these effects have not yet been proven. Thus, the biological action of PDT with chlorophyll-a on hepatic parameters related to energy metabolism and oxidative stress in T1DM Wistar rats was investigated. Evaluation of the acute effects of this pigment was performed by incubation of isolated hepatocytes with chlorophyll-a and the chronic effects were evaluated by oral treatment with chlorophyll-based extract, with post-analysis of the intact liver by in situ perfusion. In both experimental protocols, chlorophyll-a decreased hepatic glucose release and glycogenolysis rate and stimulated the glycolytic pathway in DM/PDT. In addition, there was a reduction in hepatic oxidative stress, noticeable by decreased lipoperoxidation, reactive oxygen species, and carbonylated proteins in livers of chlorophyll-treated T1DM rats. These are indicators of the potential capacity of chlorophyll-a in improving the status of the diabetic liver.


Assuntos
Clorofila/análogos & derivados , Diabetes Mellitus Experimental/tratamento farmacológico , Glicólise/efeitos dos fármacos , Fígado/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Fármacos Fotossensibilizantes/administração & dosagem , Animais , Clorofila/administração & dosagem , Diabetes Mellitus Experimental/patologia , Quimioterapia Combinada , Metabolismo Energético/efeitos dos fármacos , Glicólise/fisiologia , Fígado/patologia , Masculino , Estresse Oxidativo/fisiologia , Fotoquimioterapia , Ratos , Ratos Wistar
6.
Nat Neurosci ; 22(11): 1806-1819, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31636448

RESUMO

Prediabetes and Alzheimer's disease both increase in prevalence with age. The former is a risk factor for the latter, but a mechanistic linkage between them remains elusive. We show that prediabetic serum hyperinsulinemia is reflected in the cerebrospinal fluid and that this chronically elevated insulin renders neurons resistant to insulin. This leads to abnormal electrophysiological activity and other defects. In addition, neuronal insulin resistance reduces hexokinase 2, thus impairing glycolysis. This hampers the ubiquitination and degradation of p35, favoring its cleavage to p25, which hyperactivates CDK5 and interferes with the GSK3ß-induced degradation of ß-catenin. CDK5 contributes to neuronal cell death while ß-catenin enters the neuronal nucleus and re-activates the cell cycle machinery. Unable to successfully divide, the neuron instead enters a senescent-like state. These findings offer a direct connection between peripheral hyperinsulinemia, as found in prediabetes, age-related neurodegeneration and cognitive decline. The implications for neurodegenerative conditions such as Alzheimer's disease are described.


Assuntos
Envelhecimento/fisiologia , Ciclo Celular/fisiologia , Senescência Celular/fisiologia , Hiperinsulinismo/fisiopatologia , Resistência à Insulina/fisiologia , Neurônios/fisiologia , Animais , Morte Celular/fisiologia , Senescência Celular/efeitos dos fármacos , Quinase 5 Dependente de Ciclina/metabolismo , Potenciais Pós-Sinápticos Excitadores/fisiologia , Expressão Gênica/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Hexoquinase/metabolismo , Hiperinsulinismo/líquido cefalorraquidiano , Potenciais Pós-Sinápticos Inibidores/fisiologia , Insulina/farmacologia , Liraglutida/farmacologia , Masculino , Aprendizagem em Labirinto/fisiologia , Metformina/farmacologia , Camundongos , Neurônios/metabolismo , Fosfotransferases/metabolismo , Cultura Primária de Células , Proteínas Serina-Treonina Quinases/metabolismo , Ubiquitinação/fisiologia , beta Catenina/metabolismo
7.
Fish Shellfish Immunol ; 94: 730-738, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31580934

RESUMO

Atrazine (ATR) causes environmental problems and damages the health of fish and aquatic animals. MicroRNAs (miRNAs) play important roles in immune regulation. However, the immunotoxicity mechanism of ATR in fish lymphocytes and the role of miRNA in this process remain unclear. To further study these mechanisms, spleen lymphocytes were exposed to 20, 40 and 60 µg/ml ATR for 18 h. Fluorescence staining and flow cytometry showed that the number of necrotic lymphocytes increased after ATR exposure. Compared with the control group, the mRNA expression of miR-181-5p was inhibited and the mRNA levels of TNF-α and HK2 were increased after ATR exposure. Additionally, the NF-κB inflammatory pathway and the levels of glycometabolism-related genes were upregulated. These results suggest that ATR induces inflammation and elevates glycometabolism in lymphocytes. We further found that the mRNA levels of receptor-interacting serine-threonine kinase 1 (RIP1), receptor-interacting serine-threonine kinase 3 (RIP3), mixed lineage kinase domain-like pseudokinase (MLKL), cylindromatosis (CYLD) and Fas-Associated protein with Death Domain (FADD) and the protein levels of RIP3 and MLKL in the treatment groups were significantly increased compared to those in control group, suggesting that ATR causes lymphocyte necroptosis. We conclude that miR-181-5p plays a key role in necroptosis in carp lymphocytes exposed to ATR by downregulating the expression of HK and TNF-α, which increases the level of glycometabolism and induces the inflammatory response, respectively.


Assuntos
Atrazina/toxicidade , Carpas/imunologia , Doenças dos Peixes/imunologia , Glicólise/imunologia , Inflamação/veterinária , MicroRNAs/metabolismo , /imunologia , Animais , Relação Dose-Resposta a Droga , Doenças dos Peixes/induzido quimicamente , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Glicólise/efeitos dos fármacos , Herbicidas/toxicidade , Inflamação/induzido quimicamente , Inflamação/imunologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Baço/efeitos dos fármacos , Baço/imunologia
8.
Acta Biochim Biophys Sin (Shanghai) ; 51(11): 1114-1122, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31650167

RESUMO

Propofol is one of the most commonly used intravenous anesthetics and plays an important role in tumor suppression. In the present study, we aimed to investigate the mechanism by which propofol attenuates tumor endothelial cells (TECs) and tumor cell adhesion to inhibit tumor metastasis in vitro. Human umbilical vein endothelial cells (HUVECs) cultured in Dulbecco's modified Eagle's medium were treated with tumor conditioned medium for 24 h, followed by 4 h of treatment with or without 25 µM of propofol, 10 µM of KN93, 500 µM of MK801, or 20 µM of rapastinel. It was found that propofol inhibited TEC adhesion and the glycolysis level of TECs. Consistently, propofol inhibited the expressions of adhesion molecules (E-selectin, ICAM-1, and VCAM-1) and glycolysis proteins (GLUT1, HK2, and LDHA) in TECs. Moreover, propofol attenuated the expression of HIF-1α, the phosphorylation of AKT and Ca2+/calmodulin-dependent protein kinase II (CaMKII), and the Ca2+ concentration in TECs. MK801, an inhibitor of NMDA receptor, and KN93, an inhibitor of CaMKII, both inhibited the expressions of adhesion molecules and glycolysis proteins, in a manner similar to propofol. Additionally, rapastine, an activator of NMDA receptor, could counteract the effects of propofol. Our results indicated that propofol attenuates intracellular Ca2+ concentration, CaMKII and AKT phosphorylation, and HIF-1α expression, probably via inhibiting the NMDA receptor, thus inhibiting glycolysis and adhesion of tumor and endothelial cells.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Adesão Celular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Neoplasias Hepáticas/tratamento farmacológico , Propofol/farmacologia , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Glicólise/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo
9.
Life Sci ; 239: 116966, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31626790

RESUMO

AIMS: Enhanced aerobic glycolysis is an essential hallmark of malignant cancer. Blocking the glycolytic pathway has been suggested as a therapeutic strategy to impair the proliferation of tumor cells. Metformin, a widely used anti-diabetes drug, exhibits anti-tumor properties. However, the underlying molecular mechanism of its action linking glucose metabolism with the suppression of proliferation has not been fully clarified. MAIN METHODS: Stable isotope tracing technology and gas chromatography-mass spectrometry method were utilized to analyze the effect of metformin on glycolytic flux in HCC cells. Western blot and immunohistochemistry were utilized to analyze the expression of phosphofructokinase-1 (PFK1) and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) in HCC cells or xenograft tumor tissues. Lactate measurement and glucose uptake assay were used to analyze the level of lactate and glucose in the presence of frucose-2,6-diphosphate (F2,6BP) in HCC cells treated with metformin. KEY FINDINGS: We found that metformin significantly impaired hepatoma cell proliferation by inhibiting the glycolytic flux via PFK1 blockade. Interestingly, activation of PFK1 by F2,6BP reverses the inhibitory effect of metformin on hepatoma cell proliferation and glycolysis. Mechanistically, PFKFB3,a potent allosteric activator of PFK1, was markedly suppressed through inhibiting hypoxia-induced factor 1 (HIF-1α) accumulation mediated by metformin. SIGNIFICANCE: Taken together these data indicate that HIF-1α/PFKFB3/PFK1 regulatory axis is a vital determinant of glucose metabolic reprogramming in hepatocellular carcinoma, which gives new insights into the action of metformin in combatting liver cancer.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Glicólise/efeitos dos fármacos , Metformina/farmacologia , Animais , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclo do Ácido Cítrico , Glucose/metabolismo , Células Hep G2 , Humanos , Hipoglicemiantes , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Metformina/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosfofrutoquinase-1/metabolismo , Fosfofrutoquinase-2/metabolismo , Fosforilação , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
10.
Oxid Med Cell Longev ; 2019: 8781690, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31531187

RESUMO

Targeting aberrant metabolism is a promising strategy for inhibiting cancer growth and metastasis. Research is now geared towards investigating the inhibition of glycolysis for anticancer drug development. Betulinic acid (BA) has demonstrated potent anticancer activities in multiple malignancies. However, its regulatory effects on glycolysis and the underlying molecular mechanisms are still unclear. BA inhibited invasion and migration of highly aggressive breast cancer cells. Moreover, BA could suppress aerobic glycolysis of breast cancer cells presenting as a reduction of lactate production, quiescent energy phenotype transition, and downregulation of aerobic glycolysis-related proteins. In this study, glucose-regulated protein 78 (GRP78) was also identified as the molecular target of BA in inhibiting aerobic glycolysis. BA treatment led to GRP78 overexpression, and GRP78 knockdown abrogated the inhibitory effect of BA on glycolysis. Further studies demonstrated that overexpressed GRP78 activated the endoplasmic reticulum (ER) stress sensor PERK. Subsequent phosphorylation of eIF2α led to the inhibition of ß-catenin expression, which resulted in the inhibition of c-Myc-mediated glycolysis. Coimmunoprecipitation assay revealed that BA interrupted the binding between GRP78 and PERK, thereby initiating the glycolysis inhibition cascade. Finally, the lung colonization model validated that BA inhibited breast cancer metastasis in vivo, as well as suppressed the expression of aerobic glycolysis-related proteins. In conclusion, our study not only provided a promising drug for aerobic glycolysis inhibition but also revealed that GRP78 is a novel molecular link between glycolytic metabolism and ER stress during tumor metastasis.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Proteínas de Choque Térmico , Proteínas de Neoplasias/metabolismo , Triterpenos/farmacologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Feminino , Proteínas de Choque Térmico/antagonistas & inibidores , Proteínas de Choque Térmico/metabolismo , Humanos , Metástase Neoplásica
11.
J Agric Food Chem ; 67(38): 10637-10645, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31513389

RESUMO

Previous studies have shown that selenite, a representative of inorganic form selenium, exerts its anticancer effect by inducing apoptosis in androgen-dependent LNCaP prostate cancer cells, but few studies have determined the nature of cell death induced by selenite in metastatic androgen-refractory PC-3 cells. Our study showed that necrosis-like cell death rather than apoptosis, pyroptosis, or autophagic cell death was caused by selenite in PC-3 cells. Mechanistically, this type of cell death was caused by ATP depletion (26.28 ± 3.39 nmol/mg of control versus 9.12 ± 2.44 nmol/mg of 10 µM selenite treatment) that resulted from phosphofructokinase activity reduction (100.17 ± 0.17% of control versus 21.74 ± 6.65% of 10 µM selenite treatment). Our study also showed that ROS production is necessary for the decrease in cellular ATP levels and in phosphofructokinase activity. To our knowledge, this is the first study showing that selenite can induce necrosis-like cell death in PC-3 cells. Our findings support selenite as an effective compound for the therapy of apoptosis-resistant prostate cancer.


Assuntos
Morte Celular/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Neoplasias da Próstata/fisiopatologia , Ácido Selenioso/farmacologia , Trifosfato de Adenosina/metabolismo , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Masculino , Fosfofrutoquinases/metabolismo , Neoplasias da Próstata/metabolismo
12.
Oncol Rep ; 42(5): 2149-2158, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31545464

RESUMO

Primary refractory acute myeloid leukemia (AML) and early recurrence of leukemic cells are among the most difficult hurdles to overcome in the treatment of AML. Moreover, uncertainties surrounding the molecular mechanism underlying refractory AML pose a challenge when it comes to developing novel therapeutic drugs. However, accumulating evidence suggests a contribution of phosphatase and tensin homolog (PTEN)/protein kinase B (AKT) signaling to the development of refractory AML. To assess PTEN/AKT signaling in AML, two types of AML cell lines were evaluated, namely control HL60 cells and KG1α cells, a refractory AML cell line that is resistant to idarubicin and cytarabine (AraC) treatment. Changes in the expression level of glycolysis­ and mitochondrial oxidative phosphorylation­related genes and proteins were evaluated by reverse transcription­quantitative polymerase chain reaction and western blot analyses, respectively. The mitochondrial oxygen consumption and extracellular acidification rates were measured using an XF24 analyzer. CCK8 assay and Annexin V/PI staining were used to analyze cell viability and cellular apoptosis, respectively. The PTEN protein was found to be depleted, whereas AKT phosphorylation levels were elevated in KG1α cells compared with HL60 cells. These changes were associated with increased expression of glucose transporter 1 and hexokinase 2, and increased lactate production. AKT inhibition decreased the proliferation of KG1α cells and decreased extracellular acidification without affecting HL60 cells. Notably, AKT inhibition increased the susceptibility of KG1α cells to chemotherapy with idarubicin and AraC. Taken together, the findings of the present study indicate that activation of AKT by PTEN deficiency sustains the refractory AML status through enhancement of glycolysis and mitochondrial respiration, effects that may be rescued by inhibiting AKT activity.


Assuntos
Resistencia a Medicamentos Antineoplásicos , Leucemia Mieloide Aguda/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Linhagem Celular Tumoral , Citarabina/farmacologia , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Glicólise/efeitos dos fármacos , Células HL-60 , Humanos , Idarubicina/farmacologia , Leucemia Mieloide Aguda/tratamento farmacológico , Fosforilação Oxidativa , Fosforilação , Transdução de Sinais
13.
J Steroid Biochem Mol Biol ; 195: 105468, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31536768

RESUMO

Clinical glucocorticoid use, and diseases that produce elevated circulating glucocorticoids, promote drastic changes in body composition and reduction in whole body insulin sensitivity. Because steroid-induced diabetes is the most common form of drug-induced hyperglycemia, we investigated mechanisms underlying the recognized phenotypes associated with glucocorticoid excess. Male C57BL/6 J mice were exposed to either 100ug/mL corticosterone (cort) or vehicle in their drinking water. Body composition measurements revealed an increase in fat mass with drastically reduced lean mass during the first week (i.e., seven days) of cort exposure. Relative to the vehicle control group, mice receiving cort had a significant reduction in insulin sensitivity (measured by insulin tolerance test) five days after drug intervention. The increase in insulin resistance significantly correlated with an increase in the number of Ki-67 positive ß-cells. Moreover, the ability to switch between fuel sources in liver tissue homogenate substrate oxidation assays revealed reduced metabolic flexibility. Furthermore, metabolomics analyses revealed a decrease in liver glycolytic metabolites, suggesting reduced glucose utilization, a finding consistent with onset of systemic insulin resistance. Physical activity was reduced, while respiratory quotient was increased, in mice receiving corticosterone. The majority of metabolic changes were reversed upon cessation of the drug regimen. Collectively, we conclude that changes in body composition and tissue level substrate metabolism are key components influencing the reductions in whole body insulin sensitivity observed during glucocorticoid administration.


Assuntos
Corticosterona/farmacologia , Glucocorticoides/farmacologia , Células Secretoras de Insulina/efeitos dos fármacos , Fígado/efeitos dos fármacos , Locomoção/efeitos dos fármacos , Animais , Composição Corporal/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Dieta Hiperlipídica , Glucose/metabolismo , Glicólise/efeitos dos fármacos , Resistência à Insulina , Fígado/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Peritonite/induzido quimicamente , Peritonite/metabolismo , Tioglicolatos
14.
Exp Hematol ; 78: 46-55.e3, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31560931

RESUMO

Hexokinase II (HXKII) is a key regulator of glucose metabolism that converts glucose to glucose 6-phosphate. Furthermore, HXKII blocks mitochondria-dependent apoptosis by inhibiting the release of cytochrome c. HXKII overexpression is frequently observed in several types of cancer and confers chemoresistance to cancer cells. In the present study, we found that compared with cell lines generated from diffuse large-B-cell lymphoma (DLBCL) patients, cell lines with features of Burkitt lymphoma have higher levels of HXKII because of the activation of both c-MYC and HIF-1. Under normoxia, HXKII levels were correlated with the growth ability of each B-cell lymphoma cell line. HXKII levels were further enhanced when the B-cell lymphoma cells were cultured under hypoxia. The high levels of HXKII induced by hypoxia conferred cisplatin resistance in all tested B-cell lymphoma cell lines. The HDAC inhibitor panobinostat significantly suppressed HXKII expression under both normoxic and hypoxic conditions. Importantly, panobinostat reversed the anti-lymphoma action of cisplatin, and this effect was diminished by hypoxia. These data suggest that HXKII plays different roles, including in the regulation of glycolysis and inhibition of apoptosis, depending on its expression levels. Furthermore, inhibition of HXKII expression by panobinostat may represent a new and attractive strategy to overcome cisplatin resistance.


Assuntos
Cisplatino/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Hexoquinase/biossíntese , Linfoma Difuso de Grandes Células B , Panobinostat/farmacologia , Idoso , Idoso de 80 Anos ou mais , Hipóxia Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Glicólise/efeitos dos fármacos , Glicólise/genética , Hexoquinase/genética , Humanos , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/enzimologia , Linfoma Difuso de Grandes Células B/genética , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo
15.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 48(3): 296-302, 2019 May 25.
Artigo em Chinês | MEDLINE | ID: mdl-31496162

RESUMO

OBJECTIVE: To investigate the effects of high dose vitamin C (VC) on proliferation of breast cancer cells and to explore its mechanisms. METHODS: Human breast cancer cells Bcap37 and MDA-MB-453 were treated with VC at low dose (0.01 mmol/L), medium dose (0.10 mmol/L) and high dose (2.00 mmol/L). Cell proliferation was determined with CCK-8 assay, protein expression was evaluated by Western blot, and the secretion of lactic acid in tumor cells was detected by colorimetric method. Bcap37 cells were inoculated in nude mice, and tumor baring nude mice were intraperitoneally injected with high VC(4 g/kg, VC group, n=5)or normal saline (control group, n=5) for 24 d. Tumor weight and body weight were calculated. RESULTS: In vitro experiments demonstrated that high dose VC significantly inhibited cell proliferation in Bcap37 and MDA-MB-453 cells (all P<0.01); the expressions of Glut1 and mTOR signaling pathway-related proteins were decreased (all P<0.05); and the secretion of lactic acid was also markedly reduced (all P<0.05). In vivo experiment showed that the tumor weight was decreased in mice treated with high-dose VC as compared with control group (P<0.05), but no difference in body weights between two groups was observed. CONCLUSIONS: High dose VC may inhibit proliferation of breast cancer cells both in vitro and in vivo through reducing glycolysis and protein synthesis.


Assuntos
Ácido Ascórbico , Neoplasias da Mama , Glicólise , Biossíntese de Proteínas , Animais , Ácido Ascórbico/farmacologia , Neoplasias da Mama/tratamento farmacológico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Humanos , Camundongos , Camundongos Nus , Biossíntese de Proteínas/efeitos dos fármacos
16.
Life Sci ; 233: 116730, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31390552

RESUMO

AIMS: Dihydroartemisinin (DHA) exhibits potential anticancer activity. However, the biological functions of DHA in prostate cancer remain largely unexplored. In this study, we aim to investigate the anti-proliferative effect and glycolysis regulation of DHA on prostate cancer cell LNCaP. MAIN METHODS: Cell proliferative activity and apoptosis inducing were detected. The gene expression was detected by mRNA microarray and results were analyzed by GO and KEGG pathway database. Expressions of glycolysis key enzymes and PI3K/AKT/HIF-1α were detected by Western blot. KEY FINDINGS: Results indicated that DHA could inhibit the LNCaP cell proliferation considerably and induce cell apoptosis. mRNA microarray showed 1293 genes were upregulated and 2322 genes were downregulated. GO and KEGG enrichment analysis suggested that glycolysis pathway was correlated with DHA inhibited the proliferation on the LNCaP cell. Western blot results showed that DHA can decrease GLUT1 and regulatory enzymes of glycolytic pathway expression probably by suppressing the activity of the intracellular Akt/mTOR and HIF-1 α. SIGNIFICANCE: Experimental validation results indicate that DHA treatment can inhibit the LNCaP cell proliferation and induce apoptosis, which may be related to glycolysis inhibition.


Assuntos
Artemisininas/farmacologia , Biomarcadores Tumorais/metabolismo , Glicólise/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Neoplasias da Próstata/tratamento farmacológico , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Antimaláricos/farmacologia , Apoptose , Biomarcadores Tumorais/genética , Proliferação de Células , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Células Tumorais Cultivadas
17.
Mol Med Rep ; 20(2): 1857-1865, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31257519

RESUMO

Triple negative breast cancer (TNBC) is one of the most aggressive types of breast cancer and has a poor prognosis. Therefore, the development of novel drugs and understanding the molecular mechanisms that may contribute to the initiation and development of TNBC are urgently required. Chidamide, a histone deacetylase inhibitor, has been reported as possessing anti­cancer properties in several cancers, however, the function of chidamide in TNBC remains to be elucidated. The present study revealed that chidamide inhibited the proliferation, colony formation and migration of TNBC cells. Experiments investigating the underlying mechanism revealed that chidamide upregulated the expression of microRNA (miR)­33a­5p in TNBC cells via RT­qPCR. Luciferase reporter assay demonstrated that miR­33a­5p was bound to the 3'­untranslated region of lactate dehydrogenase A (LDHA) and decreased the expression of LDHA in TNBC cells. In addition, chidamide suppressed the expression of LDHA and significantly decreased the glycolysis of TNBC cells. Collectively, the results of the present study demonstrated that chidamide reprogramed glucose metabolism, partially by targeting the miR­33a­5p/LDHA pathway, in TNBC. These findings indicate that chidamide may be a promising novel drug in the treatment of patients with TNBC.


Assuntos
Aminopiridinas/farmacologia , Benzamidas/farmacologia , L-Lactato Desidrogenase/genética , MicroRNAs/genética , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Humanos , Transdução de Sinais/efeitos dos fármacos , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologia
18.
Plant Physiol Biochem ; 142: 143-150, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31284138

RESUMO

The current study illustrates the systemic damages caused by increasing concentration of fluoride in non-aromatic rice variety, IR-64 and aromatic rice Gobindobhog (GB). Analysis of the physiological parameters like shoot length, root length and electrolyte leakage along with crucial damage indices like chlorophyll, malondialdehyde, H2O2 and protease activity indicated higher fluoride adaptation in GB compared to IR-64. IR-64 exhibited unregulated fluoride bioaccumulation when exposed to 25 mg L-1 NaF stress, whereas fluoride uptake in GB was much regulated. Gene expression studies proposed that CLC2 rather than CLC1 mediated the fluoride import. Fluoride also triggered higher P-H+/ATPase accumulation in GB compared to IR-64, thus highlighting efficient homeostasis in stressed GB. Unlike IR-64, GB could maintain photosynthesis (RuBisCo expression), sugar metabolism (α-amylase expression and activity), glycolysis and Krebs cycle even under high concentration of fluoride stress. Fluoride also inhibited nitrate reductase activity in both the cultivars. The present research illustrates differential phytotoxicity emerging out of fluoride accumulation in rice seedlings, highlighting that IR-64 is a highly susceptible variety, whereas GB exhibits physiological plasticity and is better adapted to higher concentrations of fluoride.


Assuntos
Fluoretos/farmacocinética , Fluoretos/toxicidade , Oryza/efeitos dos fármacos , Oryza/fisiologia , Ciclo do Ácido Cítrico/fisiologia , Enzimas/genética , Enzimas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Germinação/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Nitrogênio/metabolismo , Fotossíntese/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Açúcares/metabolismo , Distribuição Tecidual
19.
Redox Biol ; 26: 101237, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31276937

RESUMO

Extracellular vesicles (EVs) generated from redox active anticancer drugs are released into the extracellular environment. These EVs contain oxidized molecules and trigger inflammatory responses by macrophages. Using a mouse model of doxorubicin (DOX)-induced tissue injury, we previously found that the major sources of circulating EVs are from heart and liver, organs that are differentially affected by DOX. Here, we investigated the effects of EVs from cardiomyocytes and those from hepatocytes on macrophage activation. EVs from H9c2 rat cardiomyocytes (H9c2 EVs) and EVs from FL83b mouse hepatocytes (FL83 b EVs) have different levels of protein-bound 4-hydroxynonenal and thus different immunostimulatory effects on mouse RAW264.7 macrophages. H9c2 EVs but not FL83 b EVs induced both pro-inflammatory and anti-inflammatory macrophage activation, mediated by NFκB and Nrf-2 pathways, respectively. DOX enhanced the effects of H9c2 EVs but not FL83 b EVs. While EVs from DOX-treated H9c2 cells (H9c2 DOXEVs) suppressed mitochondrial respiration and increased glycolysis of macrophages, EVs from DOX-treated FL83b cells (FL83b DOXEVs) enhanced mitochondrial reserve capacity. Mechanistically, the different immunostimulatory functions of H9c2 EVs and FL83 b EVs are regulated, in part, by the redox status of the cytoplasmic thioredoxin 1 (Trx1) of macrophages. H9c2 DOXEVs lowered the level of reduced Trx1 in cytoplasm while FL83b DOXEVs did the opposite. Trx1 overexpression alleviated the effect of H9c2 DOXEVs on NFκB and Nrf-2 activation and prevented the upregulation of their target genes. Our findings identify EVs as a novel Trx1-mediated redox mediator of immune response, which greatly enhances our understanding of innate immune responses during cancer therapy.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Vesículas Extracelulares/imunologia , Hepatócitos/química , Miócitos Cardíacos/química , Tiorredoxinas/imunologia , Aldeídos/imunologia , Aldeídos/metabolismo , Aldeídos/farmacologia , Animais , Linhagem Celular , Meios de Cultivo Condicionados/química , Vesículas Extracelulares/química , Regulação da Expressão Gênica , Glicólise/efeitos dos fármacos , Hepatócitos/metabolismo , Ativação de Macrófagos/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/imunologia , Mitocôndrias/metabolismo , Miócitos Cardíacos/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Oxirredução , Células RAW 264.7 , Ratos , Tiorredoxinas/genética
20.
Cell Oncol (Dordr) ; 42(5): 679-690, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31325096

RESUMO

BACKGROUND: Apatinib is a tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor-2 (VEGFR2), and has shown encouraging therapeutic effects in various malignant tumors. As yet, however, the role of apatinib in ovarian cancer has remained unknown. Here, we sought to elucidate the role of apatinib in the in vitro and in vivo viability and proliferation of ovarian cancer cells, as well as in glucose metabolism in these cells. METHODS: The effects of apatinib on ovarian cancer cell viability and proliferation were assessed using Cell Counting Kit-8 (CCK-8) and colony formation assays, respectively. The expression of VEGFR2/AKT1/SOX5/GLUT4 pathway proteins was assessed using Western blotting, and glucose uptake and lactate production assays were used to detect glycolysis in ovarian cancer cells. SOX5 was exogenously over-expressed and silenced in ovarian cancer cells using expression vector and shRNA-based methods, respectively. RNA expression analyses were performed using RNA-seq and gene-chip-based methods. GLUT4 promoter activity was assessed using a dual-luciferase reporter assay. The expression of p-VEGFR2 (Tyr1175), p-AKT1 (Ser473), p-GSK3ß (Ser9), SOX5 and GLUT4 in xenograft tissues was assessed using immunohistochemistry (IHC). RESULTS: We found that apatinib inhibited the in vitro and in vivo viability and proliferation in Hey and OVCA433 ovarian cancer cells in a dose-dependent and time-dependent manner. We also found that apatinib effectively suppressed glucose uptake and lactate production by blocking the expression of GLUT4 in these cells. In addition, we found that SOX5 predominantly rescued the inhibitory effect of apatinib on GLUT4 expression by activating its promoter. Finally, we found that apatinib regulated the expression of SOX5 by suppressing the VEGFR2/AKT1/GSK3ß signaling pathway. CONCLUSIONS: From our results, we conclude that apatinib suppresses the in vitro and in vivo viability and proliferation of ovarian cancer cells, as well as glycolysis by inhibiting the VEGFR2/AKT1/GSK3ß/SOX5/GLUT4 signaling pathway. Apatinib may serve as a promising drug for the treatment of ovarian cancer.


Assuntos
Antineoplásicos/farmacologia , Glicólise/efeitos dos fármacos , Neoplasias Ovarianas/metabolismo , Piridinas/farmacologia , Fatores de Transcrição SOXD/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Transportador de Glucose Tipo 4/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Piridinas/uso terapêutico , Fatores de Transcrição SOXD/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
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