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1.
Br J Radiol ; 92(1103): 20190198, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31538514

RESUMO

OBJECTIVE: Radiation therapy is among the most effective and widely used modalities of cancer therapy in current clinical practice. In this era of personalized radiation medicine, high-throughput data now provide the means to investigate novel biomarkers of radiation response. Large-scale efforts have identified several radiation response signatures, which poses two challenges, namely, their analytical validity and redundancy of gene signatures. METHODS: To address these fundamental radiogenomics questions, we curated a database of gene expression signatures predictive of radiation response under oxic and hypoxic conditions. RadiationGeneSigDB has a collection of 11 oxic and 24 hypoxic signatures with the standardized gene list as a gene symbol, Entrez gene ID, and its function. We present the utility of this database by gaining an understanding of hypoxia-associated miRNA by applying a penalized multivariate model; by comparing breast cancer oxic signatures in cell line data vs patient data; and by comparing the similarity of head and neck cancer hypoxia signatures at the pathway level in clinical tumour data. RESULTS: We obtained a set of miRNA highly associated both positively and negatively to the hypoxia gene signatures, across pan-cancer. In addition, we identified moderate correlations between breast cancer oxic signatures in patient data, and significant differences across molecular subtypes. Moreover, we also found that different set of pathways to be enriched using the head and neck hypoxia signatures, although, they are found to be concordant when applied on the patient data. CONCLUSION: This valuable, curated repertoire of published gene expression signatures provides motivating case studies for how to search for similarities in radiation response for tumours arising from different tissues across model systems under oxic and hypoxic conditions, and how a well-curated set of gene signatures can be used to generate novel biological hypotheses about the functions of non-coding RNA. ADVANCES IN KNOWLEDGE: We envision that RadiationSigDB database will help accelerate preclinical radiotherapeutic discovery pipelines in terms of analytical validity of novel biomarkers of radiation response and the need for ensemble approaches to clinical genomic biomarkers.


Assuntos
Neoplasias da Mama/genética , Bases de Dados Factuais , Neoplasias de Cabeça e Pescoço/genética , MicroRNAs/genética , Transcriptoma/genética , Pesquisa Biomédica , Neoplasias da Mama/radioterapia , Marcadores Genéticos/genética , Neoplasias de Cabeça e Pescoço/radioterapia , Humanos , Hipóxia/genética , Oxigênio/fisiologia , Células Tumorais Cultivadas
2.
Life Sci ; 232: 116601, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31252000

RESUMO

AIMS: Tet1, Tet2, and interleukin-6 (IL-6) have been linked to atherosclerosis. Whether Tet3 has a relationship with atherosclerosis and IL-6 was unknown. This study aims to determine the link between Tet3 and IL-6, and the role of Tet3 in prenatal hypoxia-induced atherosclerosis in offspring rats. MAIN METHODS: Pregnant rats were divided into hypoxia and control group. Their male offspring were tested at 20 months old. Hematoxylin-eosin staining and transmission electron microscopic staining were used. Gene mRNA and protein levels were measured with q-PCR or Western blotting. Cell viability and migration was tested with MTT or cell scratch assay. 5-hmC and 5-mC expression were obtained by qGlucMS-PCR; 5-hmC and 5-mC activity were obtained by dot blotting. KEY FINDINGS: Chronic prenatal hypoxia increased Tet3 and IL-6 expression, and decreased Tet3 activity in offspring rats. GlucMS-qPCR showed the percentage of 5-hmC was significantly up-regulated in the promoter of IL-6 in both the rats and cells. Moreover, 5-hmC percentage also was increased in the A7r5 cells transfected with Tet3. Furthermore, Tet3 promoted proliferation and migration of A7r5 cells. However, Tet3 was not sensitive to acute hypoxia, while influenced by HIF-1α DNA element. SIGNIFICANCE: Tet3 enhanced IL-6 expression though up-regulating 5-hmC percentage in the IL-6 promoter.


Assuntos
Aterosclerose/metabolismo , Desoxicitidina/análogos & derivados , Dioxigenases/metabolismo , Hipóxia/metabolismo , Interleucina-6/biossíntese , Animais , Aterosclerose/genética , Aterosclerose/patologia , Movimento Celular/fisiologia , Sobrevivência Celular/fisiologia , Desoxicitidina/metabolismo , Epigênese Genética , Feminino , Hipóxia/genética , Hipóxia/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Oxigenases de Função Mista/metabolismo , Gravidez , Complicações na Gravidez/metabolismo , Complicações na Gravidez/patologia , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ativação Transcricional , Regulação para Cima
3.
Nat Commun ; 10(1): 2824, 2019 06 27.
Artigo em Inglês | MEDLINE | ID: mdl-31249305

RESUMO

The fibrogenic response in tissue-resident fibroblasts is determined by the balance between activation and repression signals from the tissue microenvironment. While the molecular pathways by which transforming growth factor-1 (TGF-ß1) activates pro-fibrogenic mechanisms have been extensively studied and are recognized critical during fibrosis development, the factors regulating TGF-ß1 signaling are poorly understood. Here we show that macrophage hypoxia signaling suppresses excessive fibrosis in a heart via oncostatin-m (OSM) secretion. During cardiac remodeling, Ly6Chi monocytes/macrophages accumulate in hypoxic areas through a hypoxia-inducible factor (HIF)-1α dependent manner and suppresses cardiac fibroblast activation. As an underlying molecular mechanism, we identify OSM, part of the interleukin 6 cytokine family, as a HIF-1α target gene, which directly inhibits the TGF-ß1 mediated activation of cardiac fibroblasts through extracellular signal-regulated kinase 1/2-dependent phosphorylation of the SMAD linker region. These results demonstrate that macrophage hypoxia signaling regulates fibroblast activation through OSM secretion in vivo.


Assuntos
Fibrose/metabolismo , Hipóxia/metabolismo , Macrófagos/metabolismo , Oncostatina M/metabolismo , Animais , Antígenos Ly/genética , Antígenos Ly/metabolismo , Feminino , Fibroblastos/metabolismo , Fibrose/genética , Fibrose/patologia , Hipóxia/genética , Hipóxia/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Oncostatina M/genética , Fosforilação , Transdução de Sinais , Proteínas Smad/genética , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta1/metabolismo
4.
Gene ; 710: 354-362, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31170438

RESUMO

To investigate the effect of HSP70-hom+2437 single nucleotide polymorphisms (SNPs) on hypoxia and ischemia condition, we constructed the neuronal hypoxic injury model and the rat middle cerebral artery occlusion (MCAO) model to compare the inhibition rate of neurons and detect the infarct volume as well as the expression of related apoptotic proteins in order to explore the possible mechanisms. The neuroblastoma cells SHSY5Y were divided into the OE (transfected with the C allele) group, OEmu (transfected with the T allele) group and negative control (NC, transfected with the empty lentiviral vector CON195) group. Varying degrees of hypoxia were induced by deferoxamine (DFO). The inhibition rate of hypoxic neurons and the expression of related apoptotic proteins were detected in the three genotype groups. While in the rat MCAO model, we built five groups including the sham group, the blank control group (injected with physiological saline), the negative control group (injected with lentivirus and physiological saline), the C allele group and the T allele group (injected with lentivirus overexpressing C and T allele). The MCAO model operation was then underwent in all five groups, the infarct volume by TTC staining and the expression of related apoptotic proteins were detected after 24 h. The results in neuronal hypoxic injury model showed a significant difference in the inhibition rate between the three groups (P < 0.05), and the average inhibition rates for the OEmu, OE and NC groups were 13.2%, 19.2% and 23.3%, respectively. The inhibition rates also differed between lower and higher DFO concentrations (P < 0.05). Compared with the NC group, Bax decreased significantly in the OE and OEmu groups, whereas PI3K and HSPA1L (HSP70-hom) increased. However, the expression of Bax in the OEmu group decreased significantly more than in the OE group, whereas PI3K and HSPA1L levels showed no difference between the two groups. Corresponding with the results above, overexpressing HSP70-hom could reduce the infarct volume of ischemic injury by TTC staining in rat MCAO model and the T allele group also had less infarct volume than C allele group. Compared with the sham group, blank control group and negative control group, Bax decreased significantly in the C and T allele groups, while HSPA1L and p- AKT increased. Furthermore, the expression of Bax in the T allele group decreased significantly more than that in the C allele group, while there were no significant differences in HSPA1L and p-AKT levels between the two groups. Therefore, the overexpression of HSP70-hom+2437 could play a protective role in hypoxic neurons and ischemic brain tissue by upregulating the expression of HSPA1L and PI3K/p-AKT and downregulating the expression of BAX. The neuroprotective effect of the T allele was stronger than that of the C allele, which may be related to the strengthened downregulation of BAX.


Assuntos
Proteínas de Choque Térmico HSP70/genética , Hipóxia/genética , Infarto da Artéria Cerebral Média/genética , Neurônios/citologia , Polimorfismo de Nucleotídeo Único , Animais , Linhagem Celular , Modelos Animais de Doenças , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Hipóxia/metabolismo , Infarto da Artéria Cerebral Média/metabolismo , Masculino , Neurônios/metabolismo , Neuroproteção , Ratos , Ratos Sprague-Dawley , Regulação para Cima , Proteína X Associada a bcl-2/metabolismo
5.
Biochemistry (Mosc) ; 84(3): 263-271, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31221064

RESUMO

Mesenchymal stromal cells (MSCs) are a population of adult stem cells that modulate functional state of neighboring tissues. During cell aging, the biological activity of MSC changes, which may affect tissue homeostasis. It is known that reducing the oxygen level in vitro to physiological values typical to a particular cell niche leads to attenuation of some morphological and functional changes associated with aging. This work aimed to study gene expression in MSCs involved in response to physiological hypoxia using a replicative aging model under physiological (5%) and atmospheric (20%) oxygen in cultures. Our results show that significant reduction of proliferative activity of MSCs is observed after 20 passages (~50 cell generations). Regardless of the oxygen, in senescent cells PKM2, SERPINE1, and VEGFA were upregulated while ANKRD37, DDIT4, HIF1A, and TXNIP were downregulated. Also, ADORA2B, BNIPL, CCNG2, EGLN1, MAP3K1, MXI1, and P4HA1 were downregulated under hypoxia. The effect of oxygen was more pronounced at earlier passages both on the cellular and transcription levels. Irrespective of the passage, genes ANGPTL4, GYS1, PKM2, SERPINE1, and TP53 were downregulated under hypoxia. Also, decreased expression was observed for ADM, F10, HMOX1, P4HB, PFKL, SLC16A3 in earlier passages, and for HK2 - in later passages. Upregulation was only observed for ANKRD37, both at early and late cultures.


Assuntos
Hipóxia/genética , Células-Tronco Mesenquimais/metabolismo , Oxigênio/metabolismo , Adulto , Células Cultivadas , Senescência Celular , Feminino , Humanos , Hipóxia/metabolismo , Pessoa de Meia-Idade
6.
Biochimie ; 163: 163-170, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31201843

RESUMO

Acute myocardial infarction causes irreversible myocardial damage and is a leading cause of death and disability worldwide. Casein kinase 2 interacting protein-1 (CKIP-1) has been suggested to confer cytoprotection against various pathologic injuries. However, it remains unclear whether CKIP-1 regulates myocardial infarction-induced cardiomyocyte injury. This study aimed to explore the potential role of CKIP-1 in regulating hypoxia-induced cardiomyocyte injury and reveal the underlying mechanism. The results demonstrated that hypoxia-exposed cardiomyocytes showed lower CKIP-1 expression. CKIP-1 restoration by transfecting a CKIP-1 expression vector significantly improved viability and reduced apoptosis in hypoxia-treated cardiomyocytes. Moreover, CKIP-1 overexpression suppressed hypoxia-induced oxidative stress in cardiomyocytes. Mechanism research revealed that CKIP-1 overexpression reduced the expression of kelch-like ECH-associated protein 1 (Keap1) and increased the nuclear translocation of nuclear factor E2-related factor 2 (Nrf2), actions which resulted in an increase in the transcription of Nrf2 target genes. However, Keap1 overexpression partially reversed CKIP-1-mediated Nrf2 promotion and cardioprotection. Notably, the blockade of Nrf2 signaling also significantly abolished CKIP-1-mediated cardioprotection. Overall, our findings demonstrate that CKIP-1 alleviates hypoxia-induced cardiomyocyte injury through the up-regulation of Nrf2 antioxidant signaling via the down-regulation of Keap1, suggesting a potential role for CKIP-1 in myocardial infarction.


Assuntos
Apoptose , Proteínas de Transporte/metabolismo , Hipóxia/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Miócitos Cardíacos/metabolismo , Transdução de Sinais , Animais , Células Cultivadas , Regulação da Expressão Gênica , Hipóxia/genética , Hipóxia/fisiopatologia , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/fisiopatologia , Miócitos Cardíacos/fisiologia , Fator 2 Relacionado a NF-E2/metabolismo , Ratos
7.
J Agric Food Chem ; 67(28): 7844-7854, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31241937

RESUMO

Bladder cancer is the fourth common cancer among men and more than 70% of the bladder cancer is nonmuscle invasive bladder cancer (NMIBC). Because of its high recurrence rate, NMIBC brings to patients physical agony and high therapy costs to the patients' family and society. It is imperative to seek a natural compound to inhibit bladder cancer cell growth and prevent bladder cancer recurrence. Cell proliferation is one of the main features of solid tumor development, and the rapid tumor cell growth usually leads to hypoxia due to the low oxygen environment. In this study we found that sulforaphane, a natural chemical which was abundant in cruciferous vegetables, could suppress bladder cancer cells proliferation in hypoxia significantly stronger than in normoxia (p < 0.05): 20 µM sulforaphane inhibited bladder cancer cell proliferation by 26.1 ± 4.1% in normoxia, while it inhibited cell proliferation by 39.7 ± 5.2% in hypoxia in RT112 cells. Consistently, sulforaphane inhibited cell proliferation by 29.7 ± 4.6% in normoxia, while it inhibited cell proliferation by 48.3 ± 5.2% in hypoxia in RT4 cells. Moreover, we revealed that sulforaphane decreased glycolytic metabolism in a hypoxia microenvironment by downregulating hypoxia-induced HIF-1α and blocking HIF-1α trans-localization to the nucleus in NMIBC cell lines. This study discovered a food sourced compound inhibiting bladder cancer cells proliferation and provided experimental evidence for developing a new bladder cancer preventive and therapeutic strategy.


Assuntos
Proliferação de Células/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Hipóxia/fisiopatologia , Isotiocianatos/administração & dosagem , Neoplasias da Bexiga Urinária/fisiopatologia , Linhagem Celular Tumoral , Glicólise/efeitos dos fármacos , Humanos , Hipóxia/tratamento farmacológico , Hipóxia/genética , Hipóxia/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo
8.
Cancer Sci ; 110(8): 2368-2377, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31222863

RESUMO

Macrophages are essential inflammatory cells which regulate the features of immune reactions within tumors. Many studies have reported their regulatory roles in immunity through cytokines and cell signaling. However, relatively few studies have focused on their metabolic features and mechanisms. We aimed to determine the signaling pathway regulating cell metabolism and the mechanism related to the regulation of human tumor-associated macrophages (TAMs) in gastric cancer (GC). Tumor-infiltrated macrophages were isolated from human GC tissues using magnetic beads, gene transcription was determined by real-time PCR, protein expression was monitored using western blots, metabolites were determined using HPLC, and transcriptional regulation was analyzed by the luciferase-based reporter gene system. A significant decrease in microRNA (miR)-30c and an increase in regulated in development and DNA damage responses 1 (REDD1) were detected in human GC TAMs, the transcription of miR-30c was negatively correlated with REDD1. MicroRNA-30c expression was suppressed by hypoxia-inducible factor-1α activation and related to decreased mTOR activity as well as glycolysis in human GC TAMs. Hypoxia-regulated miR-30c downregulated REDD-1 expression by targeting its 3'UTR. Overexpression of miR-30c or restored mTOR activity in macrophages with miR-30cLow expression promoted M1 macrophage differentiation and function in TAMs. Therefore, hypoxia in the human GC microenvironment suppressed the expression of miR-30c, and decreased mTOR activity as well as glycolysis in GC TAMs, thus inhibiting M1 differentiation and function. These results provide a novel metabolic strategy for tumor microenvironment-based therapy.


Assuntos
Glicólise/genética , Hipóxia/genética , Macrófagos/patologia , MicroRNAs/genética , Neoplasias Gástricas/genética , Serina-Treonina Quinases TOR/genética , Regiões 3' não Traduzidas/genética , Diferenciação Celular , Linhagem Celular , Proliferação de Células/genética , Regulação para Baixo/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Hipóxia/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/genética , Neoplasias Gástricas/patologia , Fatores de Transcrição/genética , Transcrição Genética/genética , Microambiente Tumoral/genética
9.
Biochimie ; 163: 128-136, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31082428

RESUMO

This study aimed at the participation of lncRNA H19, endothelial NADPH oxidases (NOX4) and miR-148b-3p in hypoxia stress in human hepatic sinusoidal endothelial cells (HHSEC), and clarifying the relationship among them. The expression of lnc H19, NOX4 and miR-148b-3p in cirrhotic patients and hypoxic HHSEC were measured by RT-PCR. The nitric oxide and hydrogen peroxide content in HHSEC were determined using ultraviolet chromatometry. The protein levels of NOX4, endothelial NOS (eNOS) and phosphorylated eNOS (p-eNOS) were measured via western blotting. The interaction between NOX4 promoter and lnc H19/miR-148b-3p was measured by dual-luciferase reporter gene detection system. The present results indicated that the expressions of NOX4 mRNA and lnc H19 were increased but miR-148b-3p was decreased in both cirrhotic patients and hypoxic HHSEC. Further, hypoxia induced the up-regulation of hydrogen peroxide and the down-regulation of eNOS/NO signaling in HHSEC. And these symptoms were ameliorated by lnc H19 shRNA and miR-148b-3p mimics. But the beneficial effects of lnc H19 shRNA and miR-148b-3p mimics were further abolished by miR-148b-3p inhibitor and NOX4 over-expression, respectively. In addition, NOX4 was a direct, negatively regulated target of miR-148b-3p, and miR-148b-3p was negatively regulated by lnc H19. Collectively, lnc H19 is a negatively regulator of microRNA-148b-3p, and participate in hypoxia stress in HHSEC via positively regulating NOX4 and negatively regulating eNOS/NO signaling.


Assuntos
Células Endoteliais/metabolismo , Hipóxia/metabolismo , Cirrose Hepática/metabolismo , Fígado/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Transdução de Sinais , Idoso , Capilares/metabolismo , Capilares/fisiopatologia , Células Endoteliais/fisiologia , Regulação da Expressão Gênica , Humanos , Hipóxia/genética , Fígado/irrigação sanguínea , Fígado/fisiopatologia , Cirrose Hepática/genética , MicroRNAs/genética , Pessoa de Meia-Idade , NADPH Oxidase 4/genética , NADPH Oxidase 4/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Fisiológico
10.
PLoS Genet ; 15(4): e1007739, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30990817

RESUMO

Sleep disordered breathing (SDB)-related overnight hypoxemia is associated with cardiometabolic disease and other comorbidities. Understanding the genetic bases for variations in nocturnal hypoxemia may help understand mechanisms influencing oxygenation and SDB-related mortality. We conducted genome-wide association tests across 10 cohorts and 4 populations to identify genetic variants associated with three correlated measures of overnight oxyhemoglobin saturation: average and minimum oxyhemoglobin saturation during sleep and the percent of sleep with oxyhemoglobin saturation under 90%. The discovery sample consisted of 8,326 individuals. Variants with p < 1 × 10(-6) were analyzed in a replication group of 14,410 individuals. We identified 3 significantly associated regions, including 2 regions in multi-ethnic analyses (2q12, 10q22). SNPs in the 2q12 region associated with minimum SpO2 (rs78136548 p = 2.70 × 10(-10)). SNPs at 10q22 were associated with all three traits including average SpO2 (rs72805692 p = 4.58 × 10(-8)). SNPs in both regions were associated in over 20,000 individuals and are supported by prior associations or functional evidence. Four additional significant regions were detected in secondary sex-stratified and combined discovery and replication analyses, including a region overlapping Reelin, a known marker of respiratory complex neurons.These are the first genome-wide significant findings reported for oxyhemoglobin saturation during sleep, a phenotype of high clinical interest. Our replicated associations with HK1 and IL18R1 suggest that variants in inflammatory pathways, such as the biologically-plausible NLRP3 inflammasome, may contribute to nocturnal hypoxemia.


Assuntos
Hexoquinase/genética , Subunidade alfa de Receptor de Interleucina-18/genética , Oxiemoglobinas/metabolismo , Sono/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Moléculas de Adesão Celular Neuronais/genética , Biologia Computacional , Proteínas da Matriz Extracelular/genética , Feminino , Redes Reguladoras de Genes , Variação Genética , Estudo de Associação Genômica Ampla , Humanos , Hipóxia/sangue , Hipóxia/genética , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteínas do Tecido Nervoso/genética , Oxigênio/sangue , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Serina Endopeptidases/genética , Síndromes da Apneia do Sono/sangue , Síndromes da Apneia do Sono/genética , Adulto Jovem
11.
Life Sci ; 227: 64-73, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31004656

RESUMO

AIM: Growing evidence suggests that endothelial-mesenchymal transition (EndMT) play key roles in pulmonary arterial remodeling during pulmonary arterial hypertension (PAH), but the underlying mechanisms have yet to be fully understood. miR-27a has been shown to promote proliferation of pulmonary arterial cells during PAH, but its role in EndMT remains unexplored. This study was designed to investigate the role and underlying mechanism of miR-27a in EndMT during PAH. MAIN METHODS: Rats were exposed in hypoxia (10% O2) for 3 weeks to induce PAH, and human pulmonary artery endothelial cells (HPAECs) were exposed in hypoxia (1% O2) for 48 h to induce EndMT. Immunohistochemistry, in situ hybridization, immunofluorescence, real-time PCR and Western blot were conducted to detect the expressions of RNAs and proteins, and luciferase assay was used to verify the putative binding site of miR-27a. KEY FINDINGS: We found that hypoxia up-regulated miR-27a in the tunica intima of rat pulmonary arteries and HPAECs, and that inhibition of miR-27a suppressed hypoxia-induced EndMT. Furthermore, elevated expression of miR-27a suppressed bone morphogenetic protein (BMP) signaling by targeting Smad5, thereby lessening Id2-mediated repression of the 2 critical mediators of EndMT (Snail and Twist). SIGNIFICANCE: Our data unveiled a novel role of miR-27a in EndMT during hypoxia-induced PAH. Thus, targeting of miR-27a-related pathway may be therapeutically harnessed to treat PAH.


Assuntos
Transição Epitelial-Mesenquimal/genética , Hipertensão Pulmonar/genética , MicroRNAs/fisiologia , Animais , Técnicas de Cultura de Células , Proliferação de Células , Células Endoteliais/metabolismo , Endotélio/metabolismo , Regulação da Expressão Gênica , Humanos , Hipertensão Pulmonar/fisiopatologia , Hipóxia/genética , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Artéria Pulmonar/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Regulação para Cima , Remodelação Vascular/genética
12.
Cell Mol Biol (Noisy-le-grand) ; 65(3): 48-57, 2019 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-30942155

RESUMO

The plateau zokor (Myospalaxbaileyi) is a specialized subterranean rodent that lives on the Qinghai-Tibet Plateau, and has successfully adapted to hypoxic environment. Raised expression of vascular endothelial growth factor (VEGF) and enhanced microvessel density (MVD) in tissues enable subterranean rodents to adapt to hypoxic sealed burrows. However, the expression of VEGF is inhibited by decreases in oxygen content, which is different from what obtains in Sprague Dawley (SD)rats. Thromspondin-1(TSP-1) is the first endogenous angiogenesis inhibitor identified inp53 pathway. It has several domains that bind to different proteins which regulate cell-to-cell interactions, inhibit endothelial cell proliferation and induce endothelial cell apoptosis (anti-angiogenesis). In this study, we analyzed the coding region and the expression pattern of TSP-1 gene in plateau zokor under different oxygen partial pressures using bioinformatics and qRT-PCR, respectively. Our results showed that the base and amino acid homologies between plateau zokor and Northern Israeli blind subterranean mole rat (Nannospalaxgalili) were 95.08 and 97.61%, respectively. There were eight parallel evolution sites with Nannospalaxgalili. Evaluation by 'Sorting Tolerant From Intolerant' (SIFT) algorithm showed four sites with significant effects on the function of TSP-1. Three-dimensional (3D) structures revealed that Asp185 and Thr270 were located in the NH2 terminal domain, with Glu536 in the Type I repeat domain, and Thr1092 in the COOH terminal domain. Compared to SD rats, the polarities of these four mutation sites changed. The expression levels of TSP-1 in plateau zokor tissues increased significantly from 2 260 m(16.12kPa) to 3 300 m(14.13kPa), but there was no significant difference in TSP-1 expression in SD rats. In conclusion, due to long-term adaption to the hypoxic environment of sealed burrows, plateau zokor upregulates the expression of TSP-1 to effect anti-angiogenesis. Moreover, mutations in gene structure of TSP-1 may play an important role in inhibiting angiogenesis.


Assuntos
Evolução Molecular , Regulação da Expressão Gênica , Hipóxia/genética , Roedores/genética , Trombospondina 1/genética , Animais , DNA Mitocondrial/genética , Modelos Moleculares , Filogenia , Subunidades Proteicas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Seleção Genética , Homologia de Sequência do Ácido Nucleico , Trombospondina 1/química , Trombospondina 1/metabolismo
13.
Int J Mol Sci ; 20(8)2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991633

RESUMO

The patients with sleep apnea syndrome are exposed to intermittent hypoxia (IH) during sleep. We previously demonstrated the IH-induced up-regulation of the mRNA levels of anorexigenic peptides proopiomelanocortin (POMC), and cocaine- and amphetamine-regulated transcript (CART) in human neuronal cells. Appetite is regulated not only by the central nervous system but also by the peptides from gastrointestinal tract. Here, we investigated the effects of IH on the gene expression(s) of appetite-inhibiting gut hormones. Human enteroendocrine Caco-2 and mouse STC-1 cells were exposed to IH [64 cycles of 5 min hypoxia (1% O2) and 10 min normoxia (21% O2)] or normoxia for 24 h. Real-time RT-PCR revealed that IH significantly increased the mRNA levels of peptide YY (PYY), glucagon-like peptide-1 (GLP-1), and neurotensin (NTS) in Caco-2 and STC-1 cells. ELISA showed that the concentrations of PYY, GLP-1, and NTS in the culture medium were significantly increased by IH. The mRNA levels of PYY, GLP-1, and NTS were significantly up-regulated even in normoxia by Trichostatin A (TSA) and were significantly decreased even in IH by 5-azacytidine (5AZC), suggesting that IH increases PYY, GLP-1, and NTS mRNAs via alterations in the chromatin structure in enteroendocrine cells. IH might have an anorexigenic influence on the enteric nervous system.


Assuntos
Células Enteroendócrinas/metabolismo , Peptídeo 1 Semelhante ao Glucagon/genética , Hipóxia/genética , Neurotensina/genética , Peptídeo YY/genética , Regulação para Cima , Animais , Células CACO-2 , Hipóxia Celular , Linhagem Celular , Humanos , Camundongos , RNA Mensageiro/genética
14.
Int J Mol Sci ; 20(8)2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991659

RESUMO

Autophagy is a cytoprotective mechanism triggered in response to adverse environmental conditions. Herein, we investigated the autophagy process in the oriental river prawn (Macrobrachium nipponense) following hypoxia. Full-length cDNAs encoding autophagy-related genes (ATGs) ATG3, ATG4B, ATG5, and ATG9A were cloned, and transcription following hypoxia was explored in different tissues and developmental stages. The ATG3, ATG4B, ATG5, and ATG9A cDNAs include open reading frames encoding proteins of 319, 264, 268, and 828 amino acids, respectively. The four M. nipponense proteins clustered separately from vertebrate homologs in phylogenetic analysis. All four mRNAs were expressed in various tissues, with highest levels in brain and hepatopancreas. Hypoxia up-regulated all four mRNAs in a time-dependent manner. Thus, these genes may contribute to autophagy-based responses against hypoxia in M. nipponense. Biochemical analysis revealed that hypoxia stimulated anaerobic metabolism in the brain tissue. Furthermore, in situ hybridization experiments revealed that ATG4B was mainly expressed in the secretory and astrocyte cells of the brain. Silencing of ATG4B down-regulated ATG8 and decreased cell viability in juvenile prawn brains following hypoxia. Thus, autophagy is an adaptive response protecting against hypoxia in M. nipponense and possibly other crustaceans. Recombinant MnATG4B could interact with recombinant MnATG8, but the GST protein could not bind to MnATG8. These findings provide us with a better understanding of the fundamental mechanisms of autophagy in prawns.


Assuntos
Proteínas de Artrópodes/genética , Proteínas Relacionadas à Autofagia/genética , Autofagia , Regulação da Expressão Gênica , Palaemonidae/genética , Aclimatação , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Hipóxia Celular , Hipóxia/genética , Palaemonidae/fisiologia , Estresse Fisiológico
15.
Mech Dev ; 156: 32-40, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30936002

RESUMO

Whether the growth of embryos after a period of stunt becomes accelerated (Catch-Up Growth, CUGr), as it occurs postnatally, has rarely been examined experimentally in any class of animals. Here, hypoxia or cold of different degrees and durations caused growth retardation in chicken embryos during the first or second week of incubation. On average, on the day of removal of the growth-inhibition, the weight of the experimental groups was 73% (wet) and 61% (dry) of control embryos, while near end-incubation (embryonic day E18) their weight averaged significantly more, respectively, 80% and 84% of controls (P < 0.001). When compared as function of developmental time, the post-intervention growth of experimental embryos was faster than that of controls. The faster growth was fully accounted for by their smaller weight at end-intervention, because embryonic growth is higher the smaller the weight. Hence, their growth was appropriate for their weight, rather than for their age. In fact, out of eight different models of growth based on age and weight (wet or dry) in various combination, the model based on embryonic wet weight at end-intervention, and weight alone, was the best predictor of the embryo's post-intervention growth. The oxygen consumption of the experimental embryos during CUGr was appropriate for their weight. In conclusion, in this experimental model of CUGr, the embryo's weight at the end of a stunt could fully predict and explain the rate of growth during the post-intervention recovery period.


Assuntos
Galinhas/crescimento & desenvolvimento , Desenvolvimento Embrionário/genética , Consumo de Oxigênio/genética , Animais , Embrião de Galinha , Galinhas/genética , Temperatura Baixa/efeitos adversos , Hipóxia/genética
16.
Cell Mol Life Sci ; 76(11): 2171-2183, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30877335

RESUMO

Despite continuing advances in the development of biomacromolecules for therapeutic purposes, successful application of these often large and hydrophilic molecules has been hindered by their inability to efficiently traverse the cellular plasma membrane. In recent years, cell-penetrating peptides (CPPs) have received considerable attention as a promising class of delivery vectors due to their ability to mediate the efficient import of a large number of cargoes in vitro and in vivo. However, the lack of target specificity of CPPs remains a major obstacle to their clinical development. To address this issue, researchers have developed strategies in which chemotherapeutic drugs are conjugated to cancer targeting peptides (CTPs) that exploit the unique characteristics of the tumor microenvironment or cancer cells, thereby improving cancer cell specificity. This review highlights several of these strategies that are currently in use, and discusses how multi-component nanoparticles conjugated to CTPs can be designed to provide a more efficient cancer therapeutic delivery strategy.


Assuntos
Antineoplásicos/farmacocinética , Peptídeos Penetradores de Células/farmacocinética , Sistemas de Liberação de Medicamentos/métodos , Terapia de Alvo Molecular/métodos , Nanopartículas/química , Neoplasias/terapia , Sequência de Aminoácidos , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Permeabilidade da Membrana Celular , Peptídeos Penetradores de Células/química , Peptídeos Penetradores de Células/metabolismo , Peptídeos Penetradores de Células/farmacologia , Expressão Gênica , Humanos , Concentração de Íons de Hidrogênio , Hipóxia/genética , Hipóxia/metabolismo , Hipóxia/patologia , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/metabolismo , Nanopartículas/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Ácidos Nucleicos Peptídicos/química , Ácidos Nucleicos Peptídicos/metabolismo , Eletricidade Estática
17.
Int J Mol Sci ; 20(6)2019 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-30909368

RESUMO

BACKGROUND: Vascular endothelial injury during ischemia generates apoptotic cell death and precedes apoptosis of underlying tissues. We aimed at studying the role of extracellular adenosine triphosphate (ATP) on endothelial cells protection against hypoxia injury. METHODS: In a hypoxic model on endothelial cells, we quantified the extracellular concentration of ATP and adenosine. The expression of mRNA (ectonucleotidases, adenosine, and P2 receptors) was measured. Apoptosis was evaluated by the expression of cleaved caspase 3. The involvement of P2 and adenosine receptors and signaling pathways was investigated using selective inhibitors. RESULTS: Hypoxic stress induced a significant increase in extracellular ATP and adenosine. After a 2-h hypoxic injury, an increase of cleaved caspase 3 was observed. ATP anti-apoptotic effect was prevented by suramin, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), and CGS15943, as well as by selective A2A, A2B, and A3 receptor antagonists. P2 receptor-mediated anti-apoptotic effect of ATP involved phosphoinositide 3-kinase (PI3K), extracellular signal-regulated kinases (ERK1/2), mitoKATP, and nitric oxide synthase (NOS) pathways whereas adenosine receptor-mediated anti-apoptotic effect involved ERK1/2, protein kinase A (PKA), and NOS. CONCLUSIONS: These results suggest a complementary role of P2 and adenosine receptors in ATP-induced protective effects against hypoxia injury of endothelial. This could be considered therapeutic targets to limit the development of ischemic injury of organs such as heart, brain, and kidney.


Assuntos
Trifosfato de Adenosina/metabolismo , Apoptose , Células Endoteliais da Veia Umbilical Humana/metabolismo , Hipóxia/metabolismo , Receptores Purinérgicos P1/metabolismo , Receptores Purinérgicos P2/metabolismo , Adenosina/metabolismo , Apoptose/genética , Biomarcadores , Espaço Extracelular/metabolismo , Expressão Gênica , Humanos , Hipóxia/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Óxido Nítrico Sintase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , RNA Mensageiro/genética , Receptores Purinérgicos P1/genética , Receptores Purinérgicos P2/genética , Transdução de Sinais , Estresse Fisiológico/genética
18.
Med Sci Monit ; 25: 2009-2015, 2019 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-30880326

RESUMO

BACKGROUND Hypoxia is an important feature of solid tumors and related to a perturbed blood supply in pathophysiologies. The aim of our research was to analyze the hypoxia response and elaborate its potential functions in colorectal cancer. MATERIAL AND METHODS The lncRNAs and mRNAs expression profile were analyzed in colorectal cancer cell line SW480 by RNA sequencing, and the functions and pathways of differentially expressed genes were screened by Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis. RESULTS In this study, 77 lncRNAs and 1327 mRNAs were identified as differentially expressed. We discovered several novel lncRNAs, such as RP11-126K1.2, RP3-438O4.4, LINC01119, CTB-22K21.2, RP11-798M19.6, and RP11-2B6.3, which had not been previously reported in regulation by hypoxia. KEGG and GO analyses identified that the differentially expressed changes in mRNAs were mainly related to regulation of basic metabolic processes and gene transcription processes and were involved in several classical pathways which were linked to cancer. CONCLUSIONS Taken together, the present findings elucidate a set of differentially expressed lncRNAs and mRNAs involved in the hypoxia response process of colorectal cancer, which may serve as a candidate diagnostic biomarker and help to explain the mechanism of initial event in colorectal carcinogenesis in colorectal cancer.


Assuntos
Neoplasias Colorretais/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Moduladores da Angiogênese/metabolismo , Linhagem Celular Tumoral , China , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/genética , Ontologia Genética , Humanos , Hipóxia/genética , Análise de Sequência com Séries de Oligonucleotídeos , RNA Longo não Codificante/análise , RNA Longo não Codificante/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Análise de Sequência de RNA/métodos , Transcriptoma/genética , Hipóxia Tumoral/genética
19.
Mol Med Rep ; 19(5): 3871-3881, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864710

RESUMO

Chronic hypoxia can be observed in the heart under physiological or pathophysiological states, including embryonic development or cyanotic congenital heart disease. The aim of the present study was to examine gene expression profiles of chronically hypoxic myocardium and to explore the pathophysiological mechanisms by which the heart adapts to chronic hypoxia. Raw data from the next­generation sequencing data set GSE36761 were downloaded from the Gene Expression Omnibus database. The data set comprised 30 specimens, including 8 healthy myocardia and 22 tetralogy of Fallot (TOF) congenital cardiac malformations; only 7 original data sets of healthy myocardia were obtained, and 5/22 TOFs were excluded. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of differentially expressed genes (DEGs) were performed. Furthermore, network analysis of DEGs using Cytoscape software based on protein­protein interaction (PPI) data was also conducted. A total of 1,260 DEGs were selected, of which 926 DEGs were enriched in 83 GO biological process terms, including extracellular matrix organization, regeneration and monocyte chemotaxis. Furthermore, 406 DEGs were enriched in 13 KEGG pathways, including cytokine­cytokine receptor interaction, focal adhesion and apoptosis. PPI network analysis indicated that six hub genes with correlated degree scores >25 among nodes were identified, including G protein subunit ß4, C­C motif chemokine receptor (CCR)1, CCR2, platelet factor 4, catenin ß1 and Jun proto­oncogene (JUN). Of these, JUN was enriched in GO terms of regeneration and neuron projection regeneration, and in KEGG pathways of focal adhesion, apoptosis and Chagas disease (American trypanosomiasis). The present bioinformatics analysis of these DEGs and hub genes may provide a molecular insight to the role of diverse genes in the pathophysiology of chronically hypoxic myocardium and in myocardial adaptation to chronic hypoxia.


Assuntos
Biomarcadores/análise , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Hipóxia/genética , Miocárdio/metabolismo , Tetralogia de Fallot/genética , Estudos de Casos e Controles , Doença Crônica , Bases de Dados Factuais , Redes Reguladoras de Genes , Humanos , Hipóxia/patologia , Miocárdio/patologia , Mapas de Interação de Proteínas , Transdução de Sinais , Tetralogia de Fallot/patologia
20.
Oncol Rep ; 41(3): 1998-2008, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30747219

RESUMO

Gastric cancer (GC) is one of the most lethal diseases worldwide, but the mechanism of GC development remains elusive. In the present study, the roles of stanniocalcin­1 (STC1) in GC were investigated. It was demonstrated that overexpression of STC1 mRNA and protein were associated with poor survival of patients with GC. The expression of STC1 was enhanced in hypoxic GC cells and overexpression of STC1 facilitated cell proliferation in hypoxia but not in normoxia. Furthermore, STC1 promoted chemoresistance, migration and invasion in hypoxia. Upregulating the expression of STC1 enhanced the expression of B cell lymphoma (Bcl)­2, neural­cadherin and matrix metalloproteinase­2, whereas it reduced the levels of cytochrome c, cleaved­caspase­9, cleaved­caspase­3 and epithelial­cadherin. However, downregulation of STC1 altered the expression of these proteins in the opposite direction. Furthermore, disturbing the expression of Bcl­2 partly reversed the changes to these proteins and also the pro­proliferation, anti­apoptosis and pro­invasion potential of STC1. In vivo experiments indicated that enhanced expression of STC1 promoted tumor growth and metastasis in mice. Collectively, the results indicated that STC1 may serve an oncogenic role in hypoxic GC via dysregulating Bcl­2, indicating that STC1 may be a potential therapeutic target in the treatment of GC.


Assuntos
Movimento Celular/genética , Proliferação de Células/genética , Resistencia a Medicamentos Antineoplásicos/genética , Glicoproteínas/genética , Hipóxia/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Neoplasias Gástricas/genética , Animais , Apoptose/genética , Linhagem Celular Tumoral , Regulação para Baixo/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Hipóxia/patologia , Masculino , Metaloproteinase 2 da Matriz/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , RNA Mensageiro/genética , Estômago/patologia , Neoplasias Gástricas/patologia , Regulação para Cima/genética
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