Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.303
Filtrar
1.
Fish Shellfish Immunol ; 92: 706-711, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31276789

RESUMO

Recently, studies have shown that IκB kinase ß (IKKß), a critical kinase in the nucleus factor kappa-B (NF-κB) pathway, participates in inflammatory responses associated with unfolded protein response (UPR) and plays an important role in ER stress-induced cell death. The unfolded protein response (UPR), which is a regulatory system to restore cellular homeostasis in the endoplasmic reticulum (ER), such as oxidative stress, bacterial infection, and virus invasion. The UPR pathways have been reported to be involved in immune responses in mammals, including the classical NF-κB pathway. However, the molecular mechanism of their crosstalk remains to be elucidated. Previously, we demonstrated that IKKß also has some conserved functions between fish and human, as grass carp (Ctenopharyngodon idella) IKKß (CiIKKß) can activate NF-κB pathway. In this study, we found that CiIKKß level in nucleus was elevated under ER stress and CiIKKß can interact with grass carp X-box-binding protein 1 (CiXBP1S), a key transcription factor in UPR. Consistently, fluorescent histochemical analysis of grass carp kidney (CIK) cells indicated that CiIKKß and CiXBP1S colocalized under ER stress. Furthermore, overexpression of CiIKKß in CIK cells enhanced ER stress tolerance by regulating UPR signaling and resulted in the significant increase of cell viability.


Assuntos
Carpas/genética , Estresse do Retículo Endoplasmático , Proteínas de Peixes/genética , Regulação da Expressão Gênica/imunologia , Quinase I-kappa B/genética , Proteína 1 de Ligação a X-Box/genética , Animais , Carpas/imunologia , Núcleo Celular/genética , Sobrevivência Celular , Proteínas de Peixes/imunologia , Quinase I-kappa B/imunologia , Resposta a Proteínas não Dobradas , Proteína 1 de Ligação a X-Box/imunologia
3.
Mol Immunol ; 111: 95-105, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31048100

RESUMO

During acute lung injury, a large number of monocytes are recruited into the pulmonary tissue, which is mainly mediated by local production of monocyte chemotactic protein 1 (MCP-1). As an essential component of the lung tissues, alveolar type II epithelial cells are one of the major sources of MCP-1. Therefore, uncovering the mechanism whereby MCP-1 production is regulated in the alveolar type II cells will provide a pivotal theoretical basis for clinical intervention in acute lung injury. In the current study, we find that there is a κB binding site in the MCP-1 promoter region, and mutation of the site leads to reduced production of MCP-1 in alveolar type II epithelial cells. In contrast, overexpression of NF-κB p65 significantly increases MCP-1 expression. Furthermore, we elucidate that IKKα/ß-NF-κB p65 signaling pathway and phosphorylation of serine 534 in NF-κB p65 are required for the maximal expression of MCP-1. Also, Activator protein 1 (AP-1) site in the promoter region and JNK1/2-c-Jun signaling are required for MCP-1 generation in alveolar type II epithelial cells. Moreover, a CCAAT/enhancer-binding protein (C/EBP) element is identified in the MCP-1 promoter region through the point mutation technique, and further experiments demonstrate that both C/EBPß and C/EBPδ are involved in basic and IL-1ß-mediated MCP-1 expression. Of note, specificity protein 1-Sp1 expression is not changed in alveolar type II epithelial cells incubated with IL-1ß, but it still control MCP-1 production by binding to the consensus sequence in the promoter region. More importantly, we find that the results derived from the cell line-MLE-12 cells and primary cells are consistent. Taken together, our data provide insights into the molecular mechanism how MCP-1 expression in inflammatory alveolar type II epithelial cells is regulated at transcription level.


Assuntos
Quimiocina CCL2/genética , Células Epiteliais/metabolismo , Interleucina-1beta/genética , Fatores de Transcrição/genética , Animais , Linhagem Celular , Quinase I-kappa B/genética , Inflamação/genética , Camundongos , NF-kappa B/genética , Regiões Promotoras Genéticas/genética , Transdução de Sinais/genética , Fator de Transcrição RelA/genética , Transcrição Genética/genética
4.
In Vitro Cell Dev Biol Anim ; 55(4): 243-251, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30887211

RESUMO

This study determined the effects of miR-338-3p on osteoclast (OC) differentiation and activation. The change levels of miR-338-3p in differentiated OCs were investigated by microRNA microarray assay and quantitative real-time PCR analysis. The effects of miR-338-3p on the differentiation and activation of OCs were determined by tartrate-resistant acid phosphatase staining resorption activity assay and Western blot. Target genes of miR-338-3p were identified by target gene prediction and dual-luciferase reporter gene detection assay as well as Western blot. Results showed that miR-338-3p was markedly downregulated in differentiated OCs. miR-338-3p could inhibit the formation and absorption activity of OCs. Western blot showed that miR-338-3p could influence the change levels of OC differentiation-related proteins. Dual-luciferase reporter gene detection assay and Western blot both showed that miR-338-3p directly targeted IKKß gene. In conclusion, miR-338-3p may affect the formation and activity of OCs by targeting the IKKß gene.


Assuntos
Regulação da Expressão Gênica , Quinase I-kappa B/genética , MicroRNAs/metabolismo , Osteogênese/genética , Animais , Sequência de Bases , Diferenciação Celular/genética , Regulação para Baixo , Quinase I-kappa B/metabolismo , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , MicroRNAs/genética , Osteoclastos/citologia , Osteoclastos/metabolismo , Ligante RANK/farmacologia , Células RAW 264.7 , Fatores de Transcrição/metabolismo
5.
Life Sci ; 224: 212-221, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30914314

RESUMO

BACKGROUND: Oscillatory shear stress (OSS) occurs in areas where atherosclerosis is prevalent. Toll-like receptor 2 (TLR2) has been associated with mechanical-stress-mediated activation of signalling pathways that may lead to inflammation, apoptosis, and atherosclerosis. Nonetheless, the mechanism underlying the connection between TLR2 and OSS is not fully understood. The purpose of this study was to investigate the link between OSS and TLR2 in human umbilical vein endothelial cell (HUVECs). METHODS: Monolayer endothelial cells were stimulated or not stimulated by OSS. Protein expression was determined by western blotting and immunofluorescent staining. Endothelial function was assessed by using dihydroethidium assay, RT-PCR, immunofluorescent staining and western blotting. The carotid artery of rats was ligated for 1 week, and a section exposed to OSS was excised and analysed. RESULTS: In vitro, the expression of TLR2 in HUVECs was activated by OSS. Additionally, OSS increased apoptosis, inflammatory changes, and oxidative stress in HUVECs, and these effects were reversed by down-regulation the expression of TLR2. We proved that OSS regulates the inflammatory response of endothelial cells through the TLR2-TAK1-IKK2 pathway. In the rats with carotid artery ligation, TLR2, TAK1 and phospho-IKK2 amounts increased at the site of OSS. SIGNIFICANCE: According to our results, the OSS-mediated HUVECs injury may be associated with an increase in TLR2 expression. Accordingly, strategies designed to reduce TLR2 expression or inhibit TLR2 activation may be an effective approach to reduce the incidence of atherosclerosis.


Assuntos
Quinase I-kappa B/metabolismo , Inflamação/patologia , MAP Quinase Quinase Quinases/metabolismo , Estresse Oxidativo , Estresse Mecânico , Receptor 2 Toll-Like/metabolismo , Animais , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana , Humanos , Quinase I-kappa B/genética , Inflamação/etiologia , Inflamação/metabolismo , MAP Quinase Quinase Quinases/genética , Ratos , Ratos Sprague-Dawley , Resistência ao Cisalhamento , Receptor 2 Toll-Like/genética
6.
Int J Mol Sci ; 20(6)2019 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-30884801

RESUMO

The small GTPase Rho and its downstream effector, Rho-kinase (ROCK), regulate various cellular functions, including organization of the actin cytoskeleton, cell adhesion and migration. A pro-inflammatory lipid mediator, lysophosphatidic acid (LPA), is a potent activator of the Rho/ROCK signalling pathway and has been shown to induce the expression of chemokines and cell adhesion molecules (CAMs). In the present study, we aimed to elucidate the precise mechanism by which ROCK regulates LPA-induced expressions and functions of chemokines and CAMs. We observed that ROCK blockade reduced LPA-induced phosphorylation of IκBα and inhibited NF-κB RelA/p65 phosphorylation, leading to attenuation of RelA/p65 nuclear translocation. Furthermore, small interfering RNA-mediated ROCK isoform knockdown experiments revealed that LPA induces the expression of monocyte chemoattractant protein-1 (MCP-1) and E-selectin via ROCK2 in human aortic endothelial cells (HAECs). Importantly, we found that ROCK2 but not ROCK1 controls LPA-induced monocytic migration and monocyte adhesion toward endothelial cells. These findings demonstrate that ROCK2 is a key regulator of endothelial inflammation. We conclude that targeting endothelial ROCK2 is potentially effective in attenuation of atherosclerosis.


Assuntos
Aterosclerose/genética , Células Endoteliais/efeitos dos fármacos , Lisofosfolipídeos/farmacologia , Quinases Associadas a rho/genética , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/genética , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Aorta/citologia , Aorta/metabolismo , Aterosclerose/metabolismo , Aterosclerose/patologia , Adesão Celular/genética , Movimento Celular/efeitos dos fármacos , Quimiocina CCL2/genética , Selectina E/genética , Células Endoteliais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Quinase I-kappa B/genética , Monócitos/efeitos dos fármacos , Fosforilação , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/genética , Quinases Associadas a rho/metabolismo
7.
Nucleic Acids Res ; 47(8): 4211-4225, 2019 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-30773595

RESUMO

In PTEN-deficient prostate cancers, AKT signaling may be activated upon suppression of androgen receptor signaling. Activation of AKT as well as NF-κB signaling involves a key regulatory protein complex containing PHLPP, FKBP51 and IKKα. Here, we report a critical role of lncRNA PCAT1 in regulating the PHLPP/FKBP51/IKKα complex and progression of castration-resistant prostate cancer (CRPC). Using database queries, bioinformatic analyses, as well as RIP and RNA pull-down assays, we discovered and validated that the lncRNA-PCAT1 perturbs the PHLPP/FKBP51/IKKα complex and activates AKT and NF-κB signaling. Expression of lncRNA-PCAT1 is positively linked to CRPC progression. PCAT1 binds directly to FKBP51, displacing PHLPP from the PHLPP/FKBP51/IKKα complex, leading to activation of AKT and NF-κB signaling. Targeting PCAT1 restores PHLPP binding to FKBP1 leading to suppression of AKT signaling. Preclinical study in a mouse model of CRPC suggests therapeutic potential by targeting lncRNA PCAT1 to suppress CRPC progression. Together, the newly identified PCAT1/FKBP51/IKKα complex provides mechanistic insight in the interplay between AKT, NF-κB and AR signaling in CRPC, and the preclinical studies suggest that a novel role for PCAT1 as a therapeutic target.


Assuntos
Adenocarcinoma/genética , Regulação Neoplásica da Expressão Gênica , NF-kappa B/genética , Neoplasias de Próstata Resistentes à Castração/genética , Proteínas Proto-Oncogênicas c-akt/genética , RNA Longo não Codificante/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/mortalidade , Adenocarcinoma/patologia , Animais , Linhagem Celular Tumoral , Proliferação de Células , Conjuntos de Dados como Assunto , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Masculino , Camundongos , Camundongos Nus , NF-kappa B/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/metabolismo , Próstata/metabolismo , Próstata/patologia , Neoplasias de Próstata Resistentes à Castração/metabolismo , Neoplasias de Próstata Resistentes à Castração/mortalidade , Neoplasias de Próstata Resistentes à Castração/patologia , Ligação Proteica , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Análise de Sobrevida , Proteínas de Ligação a Tacrolimo/genética , Proteínas de Ligação a Tacrolimo/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Dis Markers ; 2019: 8418379, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30723530

RESUMO

Background: Preeclamptic pregnancies often present an intensified inflammatory state associated with the nuclear activity of NFκB. NEMO is an essential regulator of nuclear factor kappa B (NFκB) in cytoplasmic and nuclear cellular compartments. The aim of the present study is to examine the level and localization of the NEMO protein in preeclamptic and nonpreeclamptic placentas. Methods: The study includes 97 preeclamptic cases and 88 controls. NEMO distribution was analyzed immunohistochemically. Its localization in the nuclear and cytoplasmic fractions, as well as in total homogenates of placental samples, was studied by western blot and ELISA. Results: The western blot and ELISA results indicate a significant difference in NEMO concentration in the total and nuclear fractions between preeclamptic and control samples (p < 0.01 and p < 0.001, respectively). In the cytoplasmic complement, similar levels of NEMO were found in preeclamptic and control placentas. In addition, immunohistochemical staining revealed that the NEMO protein is mainly localized in the syncytiotrophoblast layer, with controls demonstrating a stronger reaction with NEMO antibodies. This study also shows that the placental level of NEMO depends on the sex of the fetus. Conclusions: The depletion of the NEMO protein in the cellular compartments of placental samples may activate one of the molecular pathways influencing the development of preeclampsia, especially in pregnancies with a female fetus. A reduction of the NEMO protein in the nuclear fraction of preeclamptic placentas may intensify the inflammatory state characteristic for preeclampsia and increase the level of apoptosis and necrosis within preeclamptic placentas.


Assuntos
Quinase I-kappa B/genética , Placenta/metabolismo , Pré-Eclâmpsia/genética , Adulto , Biomarcadores/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Quinase I-kappa B/metabolismo , Recém-Nascido , Masculino , Pré-Eclâmpsia/metabolismo , Gravidez , Fatores Sexuais
9.
Immunity ; 50(2): 348-361.e4, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30737145

RESUMO

NF-κB (nuclear factor κB) signaling is considered critical for single positive (SP) thymocyte development because loss of upstream activators of NF-κB, such as the IKK complex, arrests their development. We found that the compound ablation of RelA, cRel, and p50, required for canonical NF-κB transcription, had no impact upon thymocyte development. While IKK-deficient thymocytes were acutely sensitive to tumor necrosis factor (TNF)-induced cell death, Rel-deficient cells remained resistant, calling into question the importance of NF-κB as the IKK target required for thymocyte survival. Instead, we found that IKK controlled thymocyte survival by repressing cell-death-inducing activity of the serine/threonine kinase RIPK1. We observed that RIPK1 expression was induced during development of SP thymocytes and that IKK was required to prevent RIPK1-kinase-dependent death of SPs in vivo. Finally, we showed that IKK was required to protect Rel-deficient thymocytes from RIPK1-dependent cell death, underscoring the NF-κB-independent function of IKK during thymic development.


Assuntos
Quinase I-kappa B/metabolismo , NF-kappa B/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Timócitos/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Quinase I-kappa B/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/genética , Proteína Serina-Treonina Quinases de Interação com Receptores/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Timócitos/citologia , Timócitos/efeitos dos fármacos , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
10.
Nat Commun ; 10(1): 860, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30808860

RESUMO

Target-centric drug development strategies prioritize single-target potency in vitro and do not account for connectivity and multi-target effects within a signal transduction network. Here, we present a systems biology approach that combines transcriptomic and structural analyses with live-cell imaging to predict small molecule inhibitors of TNF-induced NF-κB signaling and elucidate the network response. We identify two first-in-class small molecules that inhibit the NF-κB signaling pathway by preventing the maturation of a rate-limiting multiprotein complex necessary for IKK activation. Our findings suggest that a network-centric drug discovery approach is a promising strategy to evaluate the impact of pharmacologic intervention in signaling.


Assuntos
NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Sistemas CRISPR-Cas , Linhagem Celular , Desenvolvimento de Medicamentos/métodos , Técnicas de Introdução de Genes , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Modelos Moleculares , Domínios e Motivos de Interação entre Proteínas/efeitos dos fármacos , Receptores Tipo I de Fatores de Necrose Tumoral/química , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Transdução de Sinais/fisiologia , Biologia de Sistemas , Fator 2 Associado a Receptor de TNF/química , Fator 2 Associado a Receptor de TNF/metabolismo , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/antagonistas & inibidores
11.
Gene ; 697: 138-143, 2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-30807779

RESUMO

In the present study, NF-κB inhibitor BAY 11-7082 and/or Hsp-27 inhibitor KRIBB-3 agents were used to investigate the molecular mechanisms mediating androgen receptor expression on prostate cancer cell lines. The decrease observed in androgen receptor and p65 expressions, particularly at 48 h, in parallel with the decrease in the phosphorylation of the p-IKK α/ß and p-Hsp-27 proteins in the LNCaP cells, indicated that androgen receptor inactivation occurred after the inhibition of the NF-κB and Hsp-27. In 22Rv1 cells, androgen receptor variant-7 was also observed to be decreased in the combined dose of 48 h. The association of this decrease with the decrease in androgen receptor and p65 expressions is a supportive result for the role of NF-κB signaling in the formation of androgen receptor variant. In androgen receptor variant-7 siRNA treatment in 22Rv1 cell lines, decrease of expression of androgen receptor variant-7 as well as decrease of expression of androgen receptor and p65 were observed. The decrease statistically significant in androgen receptor and p65 expressions was even greater when siRNA treatment was followed with low dose and time (6 h) combined treatment after transfection. We also showed that increased Noxa and decreased Bcl-2 protein level, indicated that apoptotic induction after this combination. In conclusion, inhibition of NF-κB and Hsp-27 is also important, along with therapies for androgen receptor variant-7 inhibition.


Assuntos
Proteínas de Choque Térmico HSP27/metabolismo , NF-kappa B/metabolismo , Neoplasias da Próstata/metabolismo , Receptores Androgênicos/biossíntese , Anisóis/farmacologia , Apoptose/fisiologia , Linhagem Celular Tumoral , Proteínas de Choque Térmico HSP27/antagonistas & inibidores , Proteínas de Choque Térmico HSP27/genética , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Isoxazóis/farmacologia , Masculino , NF-kappa B/genética , Nitrilos , Neoplasias da Próstata/genética , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Transdução de Sinais , Sulfonas
12.
Viruses ; 11(2)2019 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-30682859

RESUMO

Proteasome is a large protein complex, which degrades most intracellular proteins. It regulates numerous cellular processes, including the removal of misfolded or unfolded proteins, cell cycle control, and regulation of apoptosis. However, the function of proteasome subunits in viral immunity has not been well characterized. In this study, we identified PSMB1, a member of the proteasome ß subunits (PSMB) family, as a negative regulator of innate immune responses during viral infection. Knockdown of PSMB1 enhanced the RNA virus-induced cytokine and chemokine production. Overexpression of PSMB1 abolished virus-induced activation of the interferon-stimulated response element (ISRE) and interferon beta (IFNß) promoters. Mechanistically, PSMB1 inhibited the activation of RIG-I-like receptor (RLR) and Toll-like receptor 3 (TLR3) signaling pathways. PSMB1 was induced after viral infection and its interaction with IKK-ε promoted degradation of IKK-ε through the ubiquitin-proteasome system. Collectively, our study demonstrates PSMB1 is an important regulator of innate immune signaling.


Assuntos
Regulação da Expressão Gênica/imunologia , Quinase I-kappa B/metabolismo , Imunidade Inata , Complexo de Endopeptidases do Proteassoma/metabolismo , Viroses/imunologia , Linhagem Celular , Quimiocinas/imunologia , Citocinas/imunologia , Proteína DEAD-box 58/antagonistas & inibidores , Proteína DEAD-box 58/genética , Técnicas de Silenciamento de Genes , Humanos , Quinase I-kappa B/genética , Interferon Tipo I/genética , Interferon beta/antagonistas & inibidores , Complexo de Endopeptidases do Proteassoma/genética , Transdução de Sinais/imunologia , Receptor 3 Toll-Like/genética , Receptor 3 Toll-Like/metabolismo , Replicação Viral
13.
Oncogene ; 38(19): 3667-3680, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30659266

RESUMO

N6-methyladenosine (m6A) is the most abundant modification in eukaryotic messenger RNAs (mRNAs), and plays important roles in many bioprocesses. However, its functions in bladder cancer (BCa) remain elusive. Here, we discovered that methyltransferase-like 3 (METTL3), a major RNA N6-adenosine methyltransferase, was significantly up-regulated in human BCa. Knockdown of METTL3 drastically reduced BCa cell proliferation, invasion, and survival in vitro and tumorigenicity in vivo. On the other hand, overexpression of METTL3 significantly promoted BCa cell growth and invasion. Through transcriptome sequencing, m6A sequencing and m6A methylated RNA immuno-precipitation quantitative reverse-transcription polymerase chain reaction, we revealed the profile of METTL3-mediated m6A modification in BCa cells for the first time. AF4/FMR2 family member 4 (AFF4), two key regulators of NF-κB pathway (IKBKB and RELA) and MYC were further identified as direct targets of METTL3-mediated m6A modification. In addition, we showed that besides NF-κB, AFF4 binds to the promoter of MYC and promotes its expression, implying a novel multilevel regulatory network downstream of METTL3. Our results uncovered an AFF4/NF-κB/MYC signaling network operated by METTL3-mediated m6A modification and provided insight into the mechanisms of BCa progression.


Assuntos
Metiltransferases/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas Repressoras/metabolismo , Fator de Transcrição RelA/metabolismo , Fatores de Elongação da Transcrição/metabolismo , Neoplasias da Bexiga Urinária/patologia , Adenosina/análogos & derivados , Adenosina/genética , Adenosina/metabolismo , Animais , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Masculino , Metiltransferases/genética , Camundongos Endogâmicos BALB C , Transdução de Sinais , Fator de Transcrição RelA/genética , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
14.
FASEB J ; 33(2): 2782-2795, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30307764

RESUMO

Fenvalerate (FEN), a mainstream pyrethroid pesticide, was initially recommended as a low-toxicity agent for controlling agricultural and domestic pests. Despite the widespread use of FEN worldwide, little data are available on FEN-induced hepatic lesions and molecular mechanisms. In the present study, we first performed an occupational cross-sectional study on FEN factory workers and found that the levels of serum alanine aminotransferase (ALT) and total antioxidant capacity increased, whereas malondialdehyde decreased in laborers in the working areas where the levels of airborne FEN were much higher compared with the office area. The results were then confirmed by animal experiments that abnormal hepatic histology, increased ALT level, and compromised hepatic oxidative capability were observed in rats exposed to a high concentration of FEN. Furthermore, the bioinformatics analysis of gene microarray in rat liver tissue showed that FEN significantly changed the expressions of genes related to the regulation of intracellular calcium ion homeostasis and the calcium signal pathway. Finally, the functional experiments in Buffalo rat liver (BRL) cells demonstrated that FEN first activated ERK MAPK, followed by IKK and NF-κB, which triggered the transcription of genes responsible for accelerating an overload of intracellular calcium ions, prompted reactive oxygen species generation in the mitochondria, and finally, induced hepatic cellular apoptosis. The calcium signaling pathway and in particular, an overload of intracellular calcium play a critical role in this pathophysiological process via the ERK/IKK/NF-κB pathway. Our study furthers the understanding of the mechanism of FEN-induced hepatic injuries and may have implications in the prevention and control of liver diseases induced by environmental pesticides.-Qiu, L.-L., Wang, C., Yao, S., Li, N., Hu, Y., Yu, Y., Xia, R., Zhu, J., Ji, M., Zhang, Z., Wang S.-L. Fenvalerate induces oxidative hepatic lesions through an overload of intracellular calcium triggered by the ERK/IKK/NF-κB pathway.


Assuntos
Cálcio/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Nitrilos/efeitos adversos , Exposição Ocupacional/efeitos adversos , Piretrinas/efeitos adversos , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Estudos Transversais , Feminino , Perfilação da Expressão Gênica , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Inseticidas/efeitos adversos , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley
15.
Urology ; 127: 61-67, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30528714

RESUMO

OBJECTIVE: To investigate how hepatocyte cell adhesion molecule (hepaCAM) regulates cancer energy metabolism through hypoxia-inducible factor (HIF-1α) in renal cell carcinoma (RCC). MATERIALS AND METHODS: The expression of hepaCAM and HIF-1α in RCC tissue samples was examined by immunohistochemistry. Glucose consumption and lactate production assays were used to detect metabolic activity in RCC cell lines. P65 and IκB kinase (IKKß) mRNA and protein expression were detected using quantitative real-time polymerase chain reaction and western blotting, respectively. Nuclear translocation of P65 was observed by immunofluorescence staining after re-expressing hepaCAM. The luciferase reporter assay was applied to validate the transcriptional activity of HIF-1α. RESULTS: HIF-1α expression was elevated and hepaCAM suppressed in RCC compared with adjacent normal tissues. Furthermore, hepaCAM re-expression significantly decreased glycolytic metabolism in RCC cell lines, and reduced HIF-1α, IKKß, and P65 expression. The expression of HIF-1α, GLUT1, LDHA, and PKM2 were further reduced with combined hepaCAM overexpression and treatment with the NF-κB inhibitor BAY11-7082, compared to hepaCAM overexpression alone. Additionally, hepaCAM decreased the transcriptional activity of HIF-1α and blocked P65 nuclear translocation by the NF-κB pathway. CONCLUSION: Our data suggest that hepaCAM suppresses the Warburg effect via the HIF-1α/NF-κB pathway in RCC, which is a facilitating factor in hepaCAM-reduced tumorigenesis.


Assuntos
Carcinogênese/genética , Carcinoma de Células Renais/genética , Regulação Neoplásica da Expressão Gênica/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Neoplasias Renais/genética , Proteínas/genética , Análise de Variância , Biópsia por Agulha , Carcinoma de Células Renais/patologia , Proteínas de Ciclo Celular , Divisão Celular/genética , Proliferação de Células/genética , Humanos , Quinase I-kappa B/genética , Imuno-Histoquímica , Neoplasias Renais/patologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Transdução de Sinais/genética , Células Tumorais Cultivadas
16.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30487159

RESUMO

Background: Inhibitory κB kinases (IKKs) play a key role in modulating proinflammatory and growth stimulating signals through their regulation of the nuclear factor κB (NF-κB) cascade. Therefore, the level of expression of IKKs represents a viable prognostic predictor with regard to various pathological processes. The prognostic value of IKKs expression in gastric cancer remains unclear. Methods: We used the 'Kaplan-Meier plotter' (KM plotter) online database, to explore the predictive prognostic value of individual IKKs members' mRNA expression to overall survival (OS) in different clinical data including pathological staging, histology, and therapies employed. Results: Our results revealed that a higher mRNA expression of inhibitor of NF-κB kinase subunit α (IKKα) was correlated to better OS, whereas higher mRNA expression of IKKß, inhibitor of NF-κB kinase subunit γ (IKKγ), inhibitor of NF-κB kinase subunit ε (IKKε), and suppressor of IKKε (SIKE) were generally correlated to unfavorable OS in gastric cancer. Increased mRNA expression of IKKε also showed better outcomes in stage IV gastric cancer. Further a correlation between elevated levels of mRNA expression of both IKKε and SIKE was found to have favorable OS in diffuse type gastric cancer. It was also revealed that high expression of SIKE had favorable OS when treated with other adjuvant therapies, while worse OS when treated only with 5FU therapy. Conclusion: Our results suggest that mRNA expression of individual IKKs and SIKE are associated with unique prognostic significance and may act as valuable prognostic biomarkers and potential targets for future therapeutic interventions in gastric cancer.


Assuntos
Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , Quinase I-kappa B/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas Serina-Treonina Quinases/genética , RNA Mensageiro/genética , Neoplasias Gástricas/genética , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais/metabolismo , Bases de Dados Factuais , Progressão da Doença , Feminino , Humanos , Quinase I-kappa B/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Estimativa de Kaplan-Meier , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Estadiamento de Neoplasias , Prognóstico , Proteínas Serina-Treonina Quinases/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/mortalidade
17.
FEBS J ; 286(3): 523-535, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30536547

RESUMO

Fas (CD95) signalling is best known for its role in apoptosis, however, recent reports have shown it to be involved in other cellular responses as well, including inflammation. Fas and its adaptor protein FADD are known to negatively regulate LPS-induced proinflammatory responses, but their role in LPS-induced type I interferon production is unknown. Here, we demonstrate that Fas engagement on macrophages, using an agonistic Fas antibody CH11, augments LPS-induced NF-κB responses, causing increased production of TNFα, IL-8, IL-6 and IL-12. Conversely, costimulation with both LPS and CH11 causes a significant reduction in the level of interferon-beta (IFNß) production. This differential effect involves the Fas adaptor FADD because while LPS-induced IL-6 production increased in FADD-/- murine embryonic fibroblasts, LPS-induced IFNß production was significantly reduced in these cells. Overexpression of a dominant negative form of FADD (FADD-DD) inhibits LPS-induced IFNß luciferase but not LPS-induced NF-κB luciferase. In contrast, overexpression of full-length FADD inhibited LPS-induced NF-κB luciferase activation but was seen to augment LPS-induced IFNß luciferase. Moreover, FADD-DD inhibits TRIF-, TRAM-, IKKε-, TBK-1- and TRAF3-induced IFNß luciferase production, with coimmunoprecipitation experiments demonstrating an interaction between FADD and TRIF. These data identify FADD as a novel component of the noncanonical Toll-like receptor 4/IFNß signalling pathway and demonstrate that both Fas and its adaptor FADD can differentially regulate the production of LPS-induced proinflammatory cytokines and type I interferons.


Assuntos
Proteína de Domínio de Morte Associada a Fas/genética , Interferon beta/genética , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Receptor 4 Toll-Like/genética , Receptor fas/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Proteínas Adaptadoras de Transporte Vesicular/genética , Proteínas Adaptadoras de Transporte Vesicular/imunologia , Animais , Anticorpos/farmacologia , Proteína de Domínio de Morte Associada a Fas/imunologia , Regulação da Expressão Gênica , Células HEK293 , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/imunologia , Interferon beta/imunologia , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , Células Jurkat , Macrófagos/citologia , Macrófagos/imunologia , Camundongos , NF-kappa B/genética , NF-kappa B/imunologia , Cultura Primária de Células , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Células RAW 264.7 , Transdução de Sinais , Células THP-1 , Fator 3 Associado a Receptor de TNF/genética , Fator 3 Associado a Receptor de TNF/imunologia , Receptor 4 Toll-Like/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Receptor fas/antagonistas & inibidores , Receptor fas/imunologia
19.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30573529

RESUMO

Shaoyao-Gancao Decoction (SGD) has been widely used for the treatment of gynopathy. The present study aimed to evaluate the therapeutic effect and potential mechanism of SGD on hyperandrogenism in polycystic ovary syndrome (PCOS) rats. In the present work, SGD was orally administrated to the PCOS rats at the dose of 12.5, 25, and 50 g/kg/d for 14 consecutive days. UPLC-MS/MS was performed to identify the main chemical components of SGD. Body weight, ovarian weight, cystic dilating follicles, and serum levels of steroid hormones were tested to evaluate the therapeutic effect of SGD. In order to further clarify the underlying mechanism, we also measured mRNA and the protein levels of NF-κB, NF-κB p65, P-NF-κB p65, and IκB by RT-qPCR and Western blotting techniques. Our results showed that SGD treatment significantly alleviated hyperandrogenism in PCOS rats as evidenced by reduced serum levels of T and increased E2 and FSH levels. In addition, SGD effectively reduced the phosphorylation of NF-κB p65 and increased the expression of IκB. Results of the present study demonstrated that SGD could ameliorate hyperandrogenism in PCOS rats, and the potential mechanism may relate to the NF-κB pathway.


Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Hiperandrogenismo/tratamento farmacológico , NF-kappa B/genética , Síndrome do Ovário Policístico/tratamento farmacológico , Animais , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hiperandrogenismo/induzido quimicamente , Hiperandrogenismo/genética , Hiperandrogenismo/patologia , Quinase I-kappa B/genética , Letrozol/toxicidade , Síndrome do Ovário Policístico/induzido quimicamente , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/patologia , Ratos , Fator de Transcrição RelA/genética
20.
Life Sci ; 218: 147-152, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30550885

RESUMO

Osteoarthritis (OA) is an inflammatory joint disease. USP14, a deubiquitinating enzyme critical for ubiquitin-mediated proteasomal protein degradation, is implicated in inflammation regulation. However, its role and mechanism in OA are poorly understood. Here, we report that USP14 is upregulated in OA articular cartilage as well as in chondrocytes treated with IL-1ß in vitro. USP14 upregulation depends on NF-κB pathway activation, since inhibition of this pathway by ACHP, a selective inhibitor of IKK-ß, abolishes USP14 upregulation. We further show that USP14 in turn exacerbates NF-κB activation through promoting IκBα deubiquitination and degradation. Functionally, USP14 aggravates the dedifferentiation effect of IL-1ß on chondrocytes, and NF-κB inhibition remarkably reverses this effect, highlighting an important role of NF-κB in mediating USP14 function. Collectively, our data reveal a previously unidentified feed-forward loop driven by USP14 and NF-κB pathway in promoting the dedifferentiation effect of IL-1ß on chondrocytes. This mechanism might offer a useful hint for OA intervention.


Assuntos
Desdiferenciação Celular , Condrócitos/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-1beta/farmacologia , NF-kappa B/metabolismo , Osteoartrite/patologia , Ubiquitina Tiolesterase/metabolismo , Animais , Biomarcadores/metabolismo , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Estudos de Casos e Controles , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Transdução de Sinais , Ubiquitina Tiolesterase/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA