Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 85
Filter
1.
Article in Chinese | WPRIM | ID: wpr-880107

ABSTRACT

OBJECTIVE@#To investigate the expression of HSP90 in bone marrow samples of multiple myeloma (MM) patients and explore its clinical significance.@*METHODS@#Maxvision immunohistochemistry technique was used to detect the protein expression level of HSP90 76 MM patients and 29 normal healthy donors. The clinical characteristics of the patients were collected, and the correlation between the HSP90 expression and the clinical characteristics was analyzed.@*RESULTS@#The count of MM patients with positive HSP90 protein was significantly higher than that of normal healthy donor, and there were no significant correlation between HSP90 expression and age, sex, hemoglobin (Hb), creatinine (CREA), blood calcium, lactate dehydrogenase (LDH), bone marrow plasma cell proportion and MM subtypes (P>0.05), but HSP90 expression was correlated with β@*CONCLUSION@#HSP90 protein was over-expressed in MM patients, and was correlated with β


Subject(s)
Bone Marrow , HSP90 Heat-Shock Proteins , Humans , Multiple Myeloma , Prognosis , beta 2-Microglobulin
2.
Acta Physiologica Sinica ; (6): 157-166, 2020.
Article in Chinese | WPRIM | ID: wpr-827072

ABSTRACT

This study was aimed to investigate the regulatory mechanism of heat shock protein 90 (Hsp90) on transcription factor EB (TFEB) during autophagy in liver cancer cells. Human hepatocellular carcinoma cell line HepG2 was treated with Hsp90 N- and C-terminal inhibitors (STA9090 and Novobiocin), respectively. Western blot and RT-PCR were used to detect the expression levels of TFEB and autophagy-related proteins. Chromatin immunoprecipitation (ChIP) assay was used to observe the ability of Hsp90α binding to the TFEB proximal promoter region. The double-luciferase gene reporter experiment was used to determine the activity of TFEB promoter. The results showed that hypoxia induced up-regulation of TFEB protein and mRNA expression levels in the HepG2 cells. The protein expression levels of TFEB, LC3 and P62 were down-regulated significantly by either STA9090 or Novobiocin, under both normoxic and hypoxic conditions. Transfection of Hsp90α-overexpressing plasmids up-regulated TFEB protein levels in either wild-type or Hsp90α knockout HepG2 cells. Hsp90 bound to the TFEB proximal promoter region and was involved in regulating TFEB transcriptional process. Whereas both STA9090 and Novobiocin inhibited Hsp90 to bind to the TFEB proximal promoter region, and decreased the activity of TFEB promoter. These results suggest that Hsp90 promotes TFEB transcription in human hepatocellular carcinoma cells by binding to the proximal promoter region, thereby up-regulating the expression levels of autophagy-related proteins.


Subject(s)
Autophagy , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors , Metabolism , Carcinoma, Hepatocellular , Metabolism , Pathology , HSP90 Heat-Shock Proteins , Metabolism , Hep G2 Cells , Humans , Liver Neoplasms , Metabolism , Pathology , Promoter Regions, Genetic
3.
Biol. Res ; 52: 4, 2019. tab, graf
Article in English | LILACS | ID: biblio-1011407

ABSTRACT

BACKGROUND: Hematoporphyrin derivative (HPD) has a sensibilization effect in lung adenocarcinoma. This study was conducted to identify the target genes of HPD in lung adenocarcinoma. METHODS: RNA sequencing was performed using the lung adenocarcinoma cell line A549 after no treatment or treatment with X-ray or X-ray + HPD. The differentially expressed genes (DEGs) were screened using Mfuzz package by noise-robust soft clustering analysis. Enrichment analysis was carried out using "BioCloud" online tool. Protein-protein interaction (PPI) network and module analyses were performed using Cytoscape software. Using WebGestalt tool and integrated transcription factor platform (ITFP), microRNA target and transcription factor (TF) target pairs were separately predicted. An integrated regulatory network was visualized with Cytoscape software. RESULTS: A total of 815 DEGs in the gene set G1 (continuously dysregulated genes along with changes in processing conditions [untreated-treated with X-ray-X-ray + treated with HPD]) and 464 DEGs in the gene set G2 (significantly dysregulated between X-ray + HPD-treated group and untreated/X-ray-treated group) were screened. The significant module identified from the PPI network for gene set G1 showed that ribosomal protein L3 (RPL3) gene could interact with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1). TFs AAA domain containing 2 (ATAD2) and protein inhibitor of activated STAT 1 (PIAS1) were separately predicted for the genes in gene set G1 and G2, respectively. In the integrated network for gene set G2, ubiquitin-specific peptidase 25 (USP25) was targeted by miR-200b, miR-200c, and miR-429. CONCLUSION: RPL3, HSP90AA1, ATAD2, and PIAS1 as well as USP25, which is targeted by miR-200b, miR-200c, and miR-429, may be the potential targets of HPD in lung adenocarcinoma.


Subject(s)
Humans , Hematoporphyrin Derivative/pharmacology , Gene Regulatory Networks/genetics , Adenocarcinoma of Lung/genetics , Lung Neoplasms/genetics , Ribosomal Proteins/drug effects , Ribosomal Proteins/genetics , Transcription Factors , Cluster Analysis , Gene Expression Regulation, Neoplastic , Sequence Analysis, RNA , HSP90 Heat-Shock Proteins/drug effects , HSP90 Heat-Shock Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/drug effects , Small Ubiquitin-Related Modifier Proteins/genetics , MicroRNAs/metabolism , Cell Line, Tumor , DNA-Binding Proteins/drug effects , DNA-Binding Proteins/genetics , Protein Inhibitors of Activated STAT/drug effects , Protein Inhibitors of Activated STAT/genetics , Flow Cytometry , ATPases Associated with Diverse Cellular Activities/drug effects , ATPases Associated with Diverse Cellular Activities/genetics , Adenocarcinoma of Lung/drug therapy , Adenocarcinoma of Lung/radiotherapy , Lung Neoplasms/drug therapy , Lung Neoplasms/radiotherapy
4.
Araçatuba; s.n; 2019. 115 p. graf, ilus, tab.
Thesis in Portuguese | LILACS, BBO | ID: biblio-1051141

ABSTRACT

A redução da reatividade vascular à fenilefrina (PE) em aorta de ratas espontaneamente hipertensas (SHR) ao final da prenhez é dependente de maior produção e/ou maior biodisponibilidade de óxido nítrico (NO), consequente do aumento da fosforilação da enzima óxido nítrico sintase endotelial (eNOS) via PI3K/Akt. A glicosilação do tipo N-acetil-glucosamina (O-GlcNAc) é uma modificação pós-traducional que compete com a fosforilação pelos mesmos sítios de ligação nas proteínas. A O-GlcNAcilação da eNOS em serina1177 leva a redução da sua atividade enquanto a fosforilação leva a sua ativação. Além destes mecanismos, a interação da eNOS com outras proteínas é capaz de regular positiva ou negativamente a sua atividade. O objetivo deste trabalho foi analisar possíveis alterações nos mecanismos de modificação pós-traducional que controlam a ativação da eNOS os quais poderiam contribuir para maior ativação e maior biodisponibilidade de NO observada em artérias de ratas prenhes. Foram avaliados o conteúdo proteico O-GlcNAc e também expressão das enzimas que participam desta modificação, O-GlcNAc transferase (OGT) e O-GlcNAcase (OGA) por Western Blotting e a atividade da OGA por ensaio bioquímico em aorta e em artéria mesentérica (2º ou 3º ramo) de ratas não prenhes (NP) e prenhes (P), normotensas (Wistar) e SHR. Ensaios de Western Blotting foram realizados também para análise da expressão das seguintes proteínas: Cav-1, p-Cav-1, CaM e Hsp90. Realizamos a contagem do número de cavéolas endoteliais da aorta e da artéria mesentérica na presença ou ausência da metil-ß-ciclodextrina (dextrina, 10 mmol/L) por microscopia eletrônica. Em estudos funcionais, avaliamos a participação da enzima OGA, pela inibição com PugNAc (100 µmol/L) e das cavéolas, utilizando um desorganizador de cavéolas, a dextrina (10 ou 20 mmol/L), na menor reatividade vascular à PE observada em aortas de ratas P. Observamos que o conteúdo de proteínas O-GlcNAciladas estava diminuído em aorta e em leito mesentérico de ratas Wistar P e SHR P. Apesar da expressão da OGT e da OGA não estar alterada, a atividade da OGA foi aumentada em aorta e leito mesentérico de ratas Wistar P, mas, encontra-se diminuída em aorta e aumentada em leito mesentérico de SHP P. A incubação com PugNAc reverteu a reduzida reatividade à PE em aorta e artéria mesentérica de ratas Wistar P mas este efeito não foi observado em vasos SHR P, demonstrando que a OGA parece ter um papel importante na redução da O-GlcNAcilação de proteínas vasculares em Wistar P. Em vasos incubados com PugNAc, a remoção do endotélio ou a incubação com L-NAME, não alterou significativamente a reatividade à PE. Juntos estes resultados sugerem que a maior atividade da eNOS observada em vasos de Wistar P, fica prejudicada na presença do PugNAc, e depende da atividade da OGA. Como não houve alteração da resposta contrátil à PE em vasos de SHR P incubados com PugNAc, possivelmente um mecanismo diferente, envolvendo a menor atividade da OGT, ocorre nestas artérias para a redução da O-GlcNAcilação da eNOS. A desorganização das cavéolas por meio da dextrina causou aumento de contração à PE e redução de potência da ACh em aortas de Wistar NP e SHR NP, porém não houve alteração em aortas de ratas Wistar P e SHR P. A dextrina não alterou o número de cavéolas em artérias de Wistar P e SHR P quando comparado com ratas NP. SHR NP apresentam um reduzido número de cavéolas das aortas em relação a Wistar NP bem como expressão reduzida de Cav-1, p-Cav-1 e CaM. A prenhez não foi capaz de alterar a expressão da Cav-1, CaM e Hsp90 em aorta e leito mesentérico de ratas normotensas e hipertensas. Estes resultados sugerem que a prenhez não altera a expressão das proteínas Cav-1, CaM e Hsp90 e possivelmente a interação com a eNOS em aorta e artérias mesentéricas de ratas normotensas e hipertensas. Em conclusão, entre os mecanismos estudados de modificação pós-traducional da eNOS, a redução da O-GlcNAcilação da eNOS, por mecanismos que envolvem a atividade da OGA e possivelmente da OGT, favoreceria a fosforilação da eNOS e consequente maior biodisponibilidade de NO, contribuindo desta forma para modulação da resposta contrátil da PE nas artérias de ratas P(AU)


Reduction of vascular reactivity to phenylephrine (PE) in aorta of spontaneously hypertensive rats (SHR) at the end of pregnancy is dependent on higher production and/or higer bioavailability of nitric oxide (NO), as a consequence of increased endothelial nitric oxide synthase enzyme (eNOS) phosphorylation, by PI3K/Akt. Glycosylation with O-linked N-acetylglucosamine (O-GlcNAc) is a post-translational modification that competes with phosphorylation by the same binding sites in proteins. O-GlcNAcylation of eNOS on serine site leads to a reduction in its activity while eNOS phosphorylation leads to its activation. In addition to these mechanisms, the interaction of eNOS with other proteins is able to regulate positively or negatively its activity. The objective of this study was to analyze possible changes in the mechanisms of post-translational modification that control the eNOS activation, which could contribute to its the greater activation and greater bioavailability of NO observed in arteries of pregnant rats. The O-GlcNAc-protein content and also the enzymes expression that participate in this modification, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) was assessed by Western Blotting, and OGA activity were evaluated by biochemical assay in the aorta and in the artery mesenteric (2nd or 3rd branch) of non-pregnant (NP) and pregnant (P), normotensive rats (Wistar) and SHR. Western Blotting assays were also performed for expression analysis of the following proteins: Cav-1, p-Cav-1, CaM and Hsp90. We performed the counting of the number of endothelial caveolae in the aorta and the mesenteric artery in the presence or absence of methyl-ß-cyclodextrin (dextrin, 10 mmol/L) by electronic microscopy. In functional studies, we evaluated the participation of the OGA enzyme, by inhibition with PugNAc (100 µmol/L) and of the caveolae, using a caveolae disassembler, dextrin (10 or 20 mmol/L), in the reduced vascular reactivity observed in aortas or mesenteric arteries of P rats. We observed that the content of O-GlcNAcylated proteins was decreased in the aorta and in the mesenteric bed of Wistar P and SHR P rats. Although OGT and OGA expression is not altered, OGA activity was increased in the aorta and mesenteric bed of Wistar P rats but was decreased in the aorta and increased in the mesenteric bed of SHP P. Incubation with PugNAc reversed the reduced reactivity to PE in the aorta and mesenteric artery of Wistar P but this effect was not observed in SHR P arteries, demonstrating that OGA appears to play an important role in reducing O-GlcNAcylation of vascular proteins in Wistar P. In arteries incubated with PugNAc, endothelial removal or incubation with L-NAME did not significantly alter reactivity to PE. Together, these results suggest that the greater eNOS activity observed in Wistar P vessels was impaired in the presence of PugNAc, and it depends on OGA activity. As there was no change in the contractile response to PE in SHR P arteries incubated with PugNAc, possibly a different mechanism, involving the lower activity of OGT, occurs in these vessels for the reduction of O-GlcNAcylation of eNOS. Dextrin caused increased contraction of PE and decreased ACh potency in Wistar NP and SHR NP aortas, but there was no change in aortas of Wistar P and SHR P. Dextrin did not alter the number of cavelae in Wistar P and SHR P arteries compared to NP rats. SHR NP showed a lower number of caveolae than to NP Wistar as well reduced expression of Cav-1 and CaM. Pregnancy was not able to alter the expression of Cav-1, CaM and Hsp90 in the aorta and mesenteric bed of normotensive and hypertensive rats. These results suggest that pregnancy does not alter the expression of Cav-1, CaM and Hsp90 proteins and possibly interaction with eNOS in the aorta and mesenteric arteries of normotensive and hypertensive rats. In conclusion, among the studied mechanisms of post-translational modification of eNOS, the reduction of O-GlcNAcylation of eNOS, by mechanisms that involve OGA activity and possibly OGT, would favor eNOS phosphorylation and consequent greater NO bioavailability, contributing in this way for modulation of the contractile response to PE in the arteries of P rats(AU)


Subject(s)
Animals , Female , Pregnancy , Aorta , Nitric Oxide Synthase , Hypertension , Glycosylation , Calmodulin , Rats, Wistar , HSP90 Heat-Shock Proteins , Caveolin 1 , Mesenteric Arteries
5.
Rio de Janeiro; s.n; 2019. 48 p. ilus.
Thesis in Portuguese | LILACS | ID: biblio-1049763

ABSTRACT

O glioblastoma é um tumor cerebral altamente invasivo e de mau prognóstico, derivado da transformação maligna de células-tronco neurais, células precursoras de oligodendrócitos e astrócitos. As células de glioblastoma expressam o receptor de reconhecimento de padrões (PRR) do tipo toll (TLR)4, que reconhece ligantes endógenos presentes no microambiente tumoral. A ativação deste receptor induz a produção de mediadores inflamatórios envolvidos na sobrevivência, migração e invasão das células tumorais. A proteína de choque térmico (HSP do inglês, heat shock protein) 90 é superexpressa por células tumorais, sendo essencial para o crescimento tumoral. Há relatos que sugerem que a ativação do TLR4 depende da atividade da HSP90, uma vez que esta mantém a integridade do receptor durante o reconhecimento de seu ligante, assim como regula diversas proteínas envolvidas na via de sinalização deste receptor, incluindo oncoproteínas e quinases. Desta forma, o objetivo do presente estudo foi investigar o efeito da ativação de TLR4 em células de glioblastoma e o impacto da atividade da HSP90 sobre a ativação e crescimento tumoral in vitro. Dados obtidos neste trabalho demonstram que as células de glioblastoma murino GL261 expressam TLR4 funcional. A ativação deste receptor pelo seu ligante, lipopolissacarídeo (LPS), levou ao aumento da expressão da proteína ácida fibrilar glial (GFAP do inglês, glial fibrillary acidic protein) e da produção de fatores envolvidos na resposta inflamatória e no desenvolvimento tumoral, tais como interleucina (IL)-6 e -10, CCL2 (do inglês CC chemokine ligand 2) e fator de crescimento do endotélio vascular (VEGF do inglês, vascular endothelial growth factor)


A ativação de TLR4 também levou à translocação do fator nuclear κB (NFκB do inglês, factor nuclear κB) para o núcleo das células. Além disso, observamos que o inibidor seletivo da atividade de HSP90 (17-allylaminogeldanamycin, 17-AAG) modulou negativamente a produção dos mediadores inflamatórios supracitados, a translocação de NFκB, assim como reduziu proliferação celular, sugerindo que a HSP90 participa da resposta inflamatória induzida pela ativação de TLR4 e do crescimento de células GL261. Neste trabalho, demonstramos que a ativação de TLR4 em células GL261 induz a resposta inflamatória e que a inibição da atividade de HSP90 contribui para a redução deste fenômeno. Considerando a importância da inflamação para o desenvolvimento tumoral, nossos dados podem ainda contribuir para o desenvolvimento futuro de novas abordagens terapêuticas. (AU)


Subject(s)
Glioblastoma , HSP90 Heat-Shock Proteins , Toll-Like Receptor 4
6.
Yonsei Medical Journal ; : 279-286, 2018.
Article in English | WPRIM | ID: wpr-713092

ABSTRACT

PURPOSE: The mechanisms underlying repolarization abnormalities during pregnancy are not fully understood. Although maternal serotonin (5-hydroxytryptamine, 5-HT) production is an important determinant for normal fetal development in mice, its role in mothers remains unclear. We evaluated the role of serotonin in ventricular repolarization in mice hearts via 5Htr3 receptor (Htr3a) and investigated the mechanism of QT-prolongation during pregnancy. MATERIALS AND METHODS: We measured current amplitudes and the expression levels of voltage-gated K⁺ (Kv) channels in freshly-isolated left ventricular myocytes from wild-type non-pregnant (WT-NP), late-pregnant (WT-LP), and non-pregnant Htr3a homozygous knockout mice (Htr3a(−/−)-NP). RESULTS: During pregnancy, serotonin and tryptophan hydroxylase 1, a rate-limiting enzyme for the synthesis of serotonin, were markedly increased in hearts and serum. Serotonin increased Kv current densities concomitant with the shortening of the QT interval in WT-NP mice, but not in WT-LP and Htr3a(−/−)-NP mice. Ondansetron, an Htr3 antagonist, decreased Kv currents in WT-LP mice, but not in WT-NP mice. Kv4.3 directly interacted with Htr3a, and this binding was facilitated by serotonin. Serotonin increased the trafficking of Kv4.3 channels to the cellular membrane in WT-NP. CONCLUSION: Serotonin increases repolarizing currents by augmenting Kv currents. Elevated serotonin levels during pregnancy counterbalance pregnancy-related QT prolongation by facilitating Htr3-mediated Kv currents.


Subject(s)
Action Potentials/drug effects , Animals , Cell Membrane/drug effects , Disease Models, Animal , Electrocardiography , Female , HSC70 Heat-Shock Proteins/metabolism , HSP90 Heat-Shock Proteins/metabolism , Heart Ventricles/drug effects , Mice, Inbred C57BL , Mice, Knockout , Myocytes, Cardiac/drug effects , Potassium Channels/metabolism , Pregnancy , Rabbits , Rats, Sprague-Dawley , Receptors, Serotonin, 5-HT3/metabolism , Serotonin/metabolism , Serotonin 5-HT3 Receptor Agonists/pharmacology
7.
Acta Physiologica Sinica ; (6): 298-304, 2017.
Article in Chinese | WPRIM | ID: wpr-348271

ABSTRACT

Protein palmitoylation, one of post-translation modifications, refers to the addition of saturated 16-carbon palmitic acid to cysteine residues via the thioester bond. It plays key roles in various functional activities, such as the interaction, stability and location of proteins. Heat shock protein 90 (Hsp90), an important molecular chaperone, has been reported to be involved in sperm capacitation. However, it remains unclear whether protein palmitoylation exists in sperm and whether Hsp90 in sperm is palmitoylated under different physiological conditions. In this study, we examined whether the protein palmitoylation is present in mouse cauda epididymis sperm using acyl-biotin exchange method, predicted the potential palmitoylated sites of Hsp90 by the software CSS-Palm 4.0 and detected the palmitoylated Hsp90 in the mouse sperm from caput epididymis and cauda epididymis by immunoprecipitation. We found that some proteins, approximately 50, 65, 72, 85 and 130 kDa, were palmitoylated in mouse cauda epididymis sperm. Five sites in two Hsp90 isoforms were predicted to be palmitoylated. The results also showed that Hsp90 in mouse sperm was palmitoylated and its palmitoylation level was involved in different physiological conditions: the palmitoylation level of cauda epididymis sperm was higher than that of caput epididymis sperm; and the palmitoylation level after capacitation was much higher than that before capacitation. In conclusion, this study reveals that protein palmitoylation is present in mouse sperm and the palmitoylated Hsp90 is associated with different physiological conditions in sperm.


Subject(s)
Animals , Epididymis , HSP90 Heat-Shock Proteins , Metabolism , Lipoylation , Male , Mice , Palmitic Acid , Chemistry , Sperm Capacitation , Spermatozoa , Metabolism
8.
Article in English | WPRIM | ID: wpr-109786

ABSTRACT

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKα/β and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKα/β level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKα/β. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.


Subject(s)
Apoptosis , Aspirin , Cell Survival , Chickens , Heat Stress Disorders , Heat-Shock Proteins , Hot Temperature , HSP90 Heat-Shock Proteins , In Vitro Techniques , Proto-Oncogene Proteins c-akt , Reactive Oxygen Species , Transducers
9.
Article in Chinese | WPRIM | ID: wpr-272493

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of HSP90 inhibitory 17-AAG on proliferation of multiple myeloma cells and its main mechanism.</p><p><b>METHODS</b>The multiple myeloma cells U266 were treated with 17-AAG of different concentrations (200, 400, 600 and 800 nmol/L) for 24, 48, and 72 hours respectively, then the proliferation rate, expression levels of β-catenin and C-MYC protein, as well as cell cycle of U266 cells were treated with 17-AAG and were detected by MTT method, Western blot and flow cytometry, respectively.</p><p><b>RESULTS</b>The 17-AAG showed inhibitory effect on the proliferation of U266 cells in dose- and time-depetent manners (r = -0.518, P < 0.05 and r = -0.473, P < 0.05), while the culture medium without 17-AAG displayed no inhibitory effect on proliferation of U266 cells (P > 0.05). The result of culturing U266 cells for 72 hours by 17-AAG of different concentrations showed that the more high of 17-AAG concentration, the more low level of β-catenin and C-MYC proteins (P < 0.05); At same time of culture, the more high of 17-AAG concentration, the more high of cell ratio in G1 phase (P < 0.05), at same concentration of 17-AAG, the more long time of culture, the more high of cell ratio in G1 phase (P < 0.05).</p><p><b>CONCLUSION</b>The HSP90 inhibitory 17-AAG can inhibit the proliferation of multiple myeloma cells, the down-regulation of Wnt/β-catenin signaling pathway and inhibition of HSP90 expression may be the main mechnisms of 17-AAG effect.</p>


Subject(s)
Apoptosis , Benzoquinones , Pharmacology , Cell Cycle , Cell Division , Cell Line, Tumor , Cell Proliferation , Down-Regulation , HSP90 Heat-Shock Proteins , Humans , Lactams, Macrocyclic , Pharmacology , Multiple Myeloma , Metabolism , Pathology , Proto-Oncogene Proteins c-myc , Metabolism , Wnt Signaling Pathway , beta Catenin , Metabolism
10.
Journal of Experimental Hematology ; (6): 1086-1090, 2016.
Article in Chinese | WPRIM | ID: wpr-246812

ABSTRACT

<p><b>UNLABELLED</b>Objective:To explore the influence of co-inhibiting mTORC2 and HSP90 on the proliferation and apoptosis of multiple myeloma(MM) cell line U266.</p><p><b>METHODS</b>During culture, the human MM cell line U266 were treated with 20 nmol/L of rapamycin, 600 nmol/L 17-AAG, 20 nmol/L of rapamycin + 600 nmol/L 17-AGG and phosphate-buffered saline (PBS), then the growth inhibition rate, morphologic changes, apoptosis rate and the expression of caspase 3 and ATK protein in U266 cells were compared and analyzed.</p><p><b>RESULTS</b>The rapamycin and 17-AAG both could inhibit the growth of U266 cells, while the inhibitory effect of rapamycin in combination with 17-AAG on growth of U266 cells was significantly higher them that of rapamycin and 17-AAG alone and control (PBS); the apoptosis rate of U266 cells treated with rapamycin, 17-AAG and their combination was higher than that of control PBS groups, and the efficacy of 2 drug conbination was higher than that of control PBS group, and the efficacy of 2 drug combination was superior to single drug. The expression levels of caspase 3 and ATK in U266 cells treated with rapamycin, 17-AAG and their combination were higher and lower than those in control group respectively, and the efficacy of 2 drug combination was superior to signle drug. There were significant difference between them (P<0.05).</p><p><b>CONCLUSION</b>The co-inhibition of mTORC2 and HSP90 can suppress the proliferation and induce the apoptosis of MM cells.</p>


Subject(s)
Apoptosis , Benzoquinones , Caspase 3 , Cell Line, Tumor , Cell Proliferation , HSP90 Heat-Shock Proteins , Humans , Lactams, Macrocyclic , Mechanistic Target of Rapamycin Complex 2 , Multiple Myeloma , Multiprotein Complexes , Sirolimus , TOR Serine-Threonine Kinases
11.
Article in English | WPRIM | ID: wpr-110767

ABSTRACT

The protective effect of aspirin during exposure to heat stress in broiler chickens was investigated. We assayed pathological damage, expression and distribution of Hsp90 protein and hsp90 mRNA expression in chicken heart tissues after oral administration of aspirin following exposure to high temperature for varying times. Heat stress induced increases in plasma aspartate aminotransferase, creatine kinase and lactate dehydrogenase activities while causing severe heart damage, which was characterized by granular and vacuolar degeneration, nuclear shrinkage and even myocardium fragmentation in cardiac muscle fibers. After aspirin administration, myocardial cells showed fewer pathological lesions than broilers treated with heat alone. A high positive Hsp90 signal was always detected in the nuclei of myocardial cells from broilers treated with aspirin, while in myocardial cells treated with heat alone, Hsp90 in the nuclei decreased, as did that in the cytoplasm. Aspirin induced rapid and significant synthesis of Hsp90 before and at the initial phase of heat stress, and significant expression of hsp90 mRNA was stimulated throughout the experiment when compared with cells exposed to heat stress alone. Thus, specific pre-induction of Hsp90 in cardiovascular tissue was useful for resisting heat stress damage because it produced stable damage-related enzymes and fewer pathologic changes.


Subject(s)
Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Aspirin/pharmacology , Cell Nucleus/genetics , Chickens , Gene Expression Regulation/drug effects , HSP90 Heat-Shock Proteins/genetics , Hot Temperature , Myocytes, Cardiac/drug effects , Stress, Physiological/drug effects
12.
Article in Chinese | WPRIM | ID: wpr-360027

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the apoptotic effects of Hsp90 selective inhibitor 17-AAG on human leukemia HL-60 and NB4 cells and analyse its possible mechanism.</p><p><b>METHODS</b>CCK-8 assay was used to quantify the growth inhibition of cells after exposure to 17-AAG for 24 hours. Flow cytometrve with annexin V/propidium iodide staining was used to detect apoptosis of leukemia cells. Then Western blot was used to detect the activation of apoptosis related protein caspase-3 and PARP level. Gene expression profile of NB4 cells treated with 17-AAG was analyzed with real-time PCR arrays.</p><p><b>RESULTS</b>The inhibition of leukemia cell proliferation displayed a dose-dependent manner. Annexin V assay, cell cycle analysis and activation of PARP demonstrate that 17-AAG induced apoptosis leukemia cells. Real-time PCR array analysis showed that expression of 56 genes significantly up-regulated and expression of 23 genes were significantly down-regulated after 17-AAG treatment.</p><p><b>CONCLUSION</b>The 17-AAG can inhibit the proliferation and induce the apoptosis of leukemia cells. After leukemia cells are treated with 17-AAG, the significant changes of apoptosis-related genes occured, and the cell apoptosis occurs via activating apoptosis related signaling pathway.</p>


Subject(s)
Apoptosis , Benzoquinones , Pharmacology , Caspase 3 , Metabolism , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Down-Regulation , HL-60 Cells , HSP90 Heat-Shock Proteins , Humans , Lactams, Macrocyclic , Pharmacology , Leukemia , Metabolism , Poly(ADP-ribose) Polymerases , Metabolism , Real-Time Polymerase Chain Reaction , Signal Transduction , Transcriptome
13.
Article in Chinese | WPRIM | ID: wpr-273728

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.</p><p><b>METHODS</b>Hyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.</p><p><b>RESULTS</b>ROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).</p><p><b>CONCLUSION</b>The intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.</p>


Subject(s)
HSP90 Heat-Shock Proteins , Metabolism , Hep G2 Cells , Hot Temperature , Humans , Proteasome Endopeptidase Complex , Metabolism , RNA, Small Interfering , Genetics , Reactive Oxygen Species , Metabolism , Up-Regulation
14.
Rev. latinoam. enferm ; 23(2): 329-336, Feb-Apr/2015. graf
Article in English | LILACS, BDENF | ID: lil-747176

ABSTRACT

OBJECTIVE: to analyze the socio-familial and community inclusion and social participation of people with disabilities, as well as their inclusion in occupations in daily life. METHOD: qualitative study with data collected through open interviews concerning the participants' life histories and systematic observation. The sample was composed of ten individuals with acquired or congenital disabilities living in the region covered by a Family Health Center. The social conception of disability was the theoretical framework used. Data were analyzed according to an interpretative reconstructive approach based on Habermas' Theory of Communicative Action. RESULTS: the results show that the socio-familial and community inclusion of the study participants is conditioned to the social determinants of health and present high levels of social inequality expressed by difficult access to PHC and rehabilitation services, work and income, education, culture, transportation and social participation. CONCLUSION: there is a need to develop community-centered care programs in cooperation with PHC services aiming to cope with poverty and improve social inclusion. .


OBJETIVO: analisar a inclusão sociofamiliar e comunitária e a participação social de pessoas com deficiência, bem como sua inserção em ocupações na vida cotidiana. MÉTODO: estudo qualitativo, com coleta de dados por meio de entrevistas abertas sobre história de vida e observação sistemática. A amostra foi composta por dez pessoas com deficiência, adquirida ou congênita, moradoras de região adstrita a um Núcleo de Saúde da Família. O referencial teórico foi a concepção social da deficiência. Os dados foram analisados segundo abordagem interpretativa reconstrutiva, fundamentada no referencial da Teoria da Ação Comunicativa de Habermas. RESULTADOS: os resultados evidenciaram que a inclusão sociofamiliar e comunitária dos sujeitos do estudo condiciona-se a determinantes sociais da saúde, apresentando índices de iniquidades sociais, expressos pela dificuldade de acesso a serviços de Atenção Primária à Saúde e de reabilitação, trabalho e renda, educação, cultura, transporte e participação social. CONCLUSÃO: conclui-se a necessidade da elaboração de programas de atenção centrados na comunidade, voltados ao enfrentamento da pobreza e à inclusão social, em articulação com serviços de Atenção Primaria à Saúde. .


OBJETIVO: analizar la inclusión social familiar y comunitaria, y la participación social de personas con deficiencia, así como su inserción en ocupaciones en la vida cotidiana. MÉTODO: estudio cualitativo, con recolección de datos por medio de entrevistas abiertas sobre historia de vida y por observación sistemática. La muestra estuvo compuesta por diez personas con deficiencia, adquirida o congénita, habitantes de una región adscrita a un Núcleo de Salud de la Familia. El referencial teórico fue la concepción social de la deficiencia. Los datos fueron analizados según abordaje interpretativo reconstructivo, fundamentado en el referencial de la Teoría de la Acción Comunicativa de Habermas. RESULTADOS: los resultados evidenciaron que la inclusión social familiar y comunitaria de los sujetos del estudio se condiciona a determinantes sociales de la salud, presentando índices de iniquidades sociales, expresados por la dificultad de acceso a servicios de Atención Primaria de la Salud y de rehabilitación, trabajo y renta, educación, cultura, transporte y participación social. CONCLUSIÓN: se concluye que existe la necesidad de elaborar programas de atención centrados en la comunidad, dirigidos al enfrentamiento de la pobreza y a la inclusión social, en articulación con servicios de Atención Primaria a la Salud. .


Subject(s)
Humans , Animals , Female , Mice , Antineoplastic Agents/pharmacology , HSP90 Heat-Shock Proteins/metabolism , Lactones/pharmacology , /drug therapy , Oximes/pharmacology , Cell Line, Tumor , Cell Proliferation , HSP90 Heat-Shock Proteins/antagonists & inhibitors , Mice, Nude , Mice, Transgenic , /metabolism , Proteolysis , Transcriptome/drug effects , Xenograft Model Antitumor Assays
15.
Cad. saúde pública ; 31(3): 620-632, 03/2015. tab
Article in Portuguese | LILACS | ID: lil-744835

ABSTRACT

O estudo descreve os pontos de venda de alimentos e sua associação com sobrepeso/obesidade em escolares de Florianópolis, Santa Catarina, Brasil. Desenho transversal com amostra probabilística de 2.506 escolares de escolas públicas (n = 19) e privadas (n = 11). O sobrepeso/obesidade foi classificado pela referência da Organização Mundial da Saúde de 2007. Foram realizadas análises brutas e ajustadas por meio de regressão de Poisson. A prevalência de sobrepeso/obesidade foi de 34,2%. Na rede pública, foram verificados 19,6% de sobrepeso e 13,5% de obesidade. Na rede privada, observaram-se 22,4% de sobrepeso e 11,1% de obesidade. Na rede pública, foi encontrada associação entre sobrepeso/obesidade e utilização da padaria (p = 0,004). Na rede privada, observou-se que os escolares de famílias que utilizaram o supermercado apresentaram 26% menos de sobrepeso/obesidade do que os escolares que não utilizam esses pontos de venda de alimentos (p = 0,003). Os dados encontrados evidenciam a existência de associação entre a utilização de alguns tipos de pontos de venda de alimentos (supermercado e padaria) e a prevalência de sobrepeso/obesidade na população escolar.


The study analyzes retail food outlets and their association with overweight/obesity in schoolchildren from Florianópolis, Santa Catarina State, Brazil. The study used a cross-sectional design with a random sample of 2,506 schoolchildren from public (n = 19) and private schools (n = 11). Overweight and obesity were classified according to World Health Organization guidelines for 2007, and crude and adjusted analyses were performed using Poisson regression. Prevalence of overweight/obesity was 34.2%. In public schools, 19.6% of the children were overweight and 13.5% were obese, as compared to 22.4% and 11.1% in private schools. An association was found in the public school system between overweight/obesity and the use of bakeries for food purchases (p = 0.004). In the private school system, children of families that bought groceries at the supermarket showed 26% less overweight/obesity compared to those who did not (p = 0.003). The data show an association between some types of food outlets (supermarkets and bakeries) and prevalence of overweight/obesity in the school-age population.


El estudio describe los puntos de venta de alimentos y su asociación con el sobrepeso/obesidad en escolares de Florianópolis, Santa Catarina, Brasil. Se trata de un estudio transversal con una muestra aleatoria de 2.506 escolares de las escuelas públicas (n = 19) y privadas (n = 11). El sobrepeso/obesidad se clasifica, en función de la OMS en 2007, con análisis ajustados y crudos que se realizaron mediante la regresión de Poisson. La prevalencia de sobrepeso/obesidad fue de un 34,2%. En el sistema público el resultado fue de un 19,6% sobrepeso y un 13,5% obesidad. En el privado se observó un 22,4% de sobrepeso y 11,1% obesidad. En el primero se encontró una correlación entre el sobrepeso/obesidad y el consumo de bollería (p = 0,004). En las escuelas privadas se observó que los escolares de familias que habían utilizado el supermercado tenían un 26% menos de sobrepeso/ obesidad que los niños en edad escolar que no utilizaron este punto de venta de alimentos (p = 0,003). En el momento del estudio existe una asociación entre el uso de algunos tipos de punto de venta de alimentos (supermercado y panadería) y la prevalencia de sobrepeso/obesidad en escolares.


Subject(s)
DNA, Fungal/chemistry , HSP90 Heat-Shock Proteins/metabolism , Nucleic Acid Conformation , Saccharomyces cerevisiae , Saccharomyces cerevisiae Proteins/metabolism , Telomere/chemistry , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Chromosomal Proteins, Non-Histone/genetics , Chromosomal Proteins, Non-Histone/metabolism , DNA, Fungal/metabolism , Enzyme Activation , HSP90 Heat-Shock Proteins/genetics , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Telomerase/metabolism , Telomere-Binding Proteins/genetics , Telomere-Binding Proteins/metabolism , Telomere/metabolism
16.
Journal of Lasers in Medical Sciences. 2015; 6 (1): 17-21
in English | IMEMR | ID: emr-160035

ABSTRACT

Heat shock proteins [HSPs] are molecular chaperones involved in protein folding, stability and turnover, and due to their role in cancer progression, the effect of low power laser irradiation [LPLI] on the expression of HSP70 and HSP90 in Jurkat E6.1 T-lymphocyte leukemia [JELT] cell line was investigated in vitro. JETL cells were irradiated with LPLI at 635nm and 780m wavelengths [energy density 9.174 J/cm[2], and assessed for the expression of HSP70 and HSP90 by flow cytometry after 24, 48 and 72 incubation time periods [ITPs]. At 24 hours ITP post-irradiation, control cultures showed that 10.7% of cells expressed HSP70, while LPLI cultures at 635nm and 780nm manifested a higher expression [32.1and 21.3%, respectively], and the difference was significant [P

Subject(s)
Humans , Male , HSP70 Heat-Shock Proteins/radiation effects , HSP90 Heat-Shock Proteins/radiation effects , Low-Level Light Therapy , Jurkat Cells , Leukemia, Lymphoid , Leukemia , T-Lymphocytes , Cell Line
17.
Chinese Journal of Oncology ; (12): 890-898, 2015.
Article in Chinese | WPRIM | ID: wpr-304480

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the antitumor efficacy and mechanism of HSP90 inhibitor FW-04-806 against Bcr/Abl(+) leukemia K562 and HL60 cells and their mechanisms of action.</p><p><b>METHODS</b>MTT assay was used to assess the proliferation-inhibiting effect of FW-04-806. Cell cycle was analyzed with propidium iodide by flow cytometry. Cell apoptosis was determined using the FITC mV apoptosis detection kit. Western blot was applied to reveal the protein expression of related proliferative and apoptotic signaling pathways. The changes of mitochondrial membrane potential were detected by flow cytometry. Protein-protein interactions was shown by co-immunoprecipitation. The level of mRNA was assessed by real-time RT-PCR.</p><p><b>RESULTS</b>FW-04-806 obviously inhibited cell proliferation in the HL60, K562 and HL60/Bcr-Abl cell lines, with an IC50 of (30.89 ± 0.12) µmol/L, (9.76 ± 0.19) µmol/L and (8.03 ± 0.26) µmol/L, respectively (P<0.001). Compared with the vehicle group, the two increasing doses of FW-04-806 showed inhibition of tumor growth at a rate of (17.40 ± 0.34)% and (34.33 ± 5.00)%, respectively, in the K562 cell line groups (P=0.003), and (18.90 ± 1.45)% and (35.60 ± 3.55)% (P=0.001) in the HL60/Bcr-Abl cell line groups. FW-04-806 dissociated Hsp90/Cdc37 chaperon/co-chaperon complex, followed by degradation of the Hsp90 proteins through proteasome pathway without affecting mRNA expression. FW-04-806 induced apoptosis and led to G2/M arrest.</p><p><b>CONCLUSION</b>Our findings indicate that FW-04-806 displays potential antitumor effect by suppressing the proliferation and apoptosis in Bcr/Abl(+) leukemia cells in vivo.</p>


Subject(s)
Apoptosis , Cell Cycle , Cell Proliferation , Fusion Proteins, bcr-abl , HL-60 Cells , HSP90 Heat-Shock Proteins , Humans , K562 Cells , Leukemia , Drug Therapy , Metabolism , Pathology , Membrane Potential, Mitochondrial , Oxazoles , Pharmacology , RNA, Messenger , Metabolism , Signal Transduction
18.
Chinese Journal of Hematology ; (12): 780-784, 2015.
Article in Chinese | WPRIM | ID: wpr-296153

ABSTRACT

<p><b>OBJECTIVE</b>To explore apoptosis of multiple myeloma (MM) cells and its mechanism by the combined inhibition of mTORC2 signaling pathway and heat shock protein 90.</p><p><b>METHODS</b>The effects of Rapamycin, 17-AAG and the combination on proliferation of MM cell lines U266 and KM3 were assessed using MTT at different time points (0, 8, 24, 48 hour). Cell apoptosis and cell cycle distribution were measured by flow cytometry. The specific proteins p-AKT (ser473), p-AKT (thr450), p-S6 (S235/236) and AKT were detected by Western blotting.</p><p><b>RESULTS</b>Rapamycin, 17- AAG and the combination suppressed the proliferation of MM cell lines U266 and KM3, especially the combination of Rapamycin and 17-AAG synergistically inhibited the proliferation (P<0.05); Rapamycin induced G1 arrest both at 24 and 48 hours, 17-AAG also induced G1 arrest, especially at 48 hours (P<0.01); Rapamycin, 17-AAG alone decreased the expression of AKT and induced MM cell apoptosis to some extent (P<0.01); Chronic rapamycin treatment inhibited mTORC2; Inhibition of both mTORC2 and chaper on pathways degraded AKT and induced MM cell apoptosis, which was significantly higher than that of any single agent (P<0.01).</p><p><b>CONCLUSION</b>Inhibition of both mTORC2 and chaper on pathways decreased the expression of AKT to induce apoptosis of MM cells in vitro.</p>


Subject(s)
Apoptosis , Benzoquinones , Pharmacology , Cell Cycle , Cell Division , Cell Line, Tumor , HSP90 Heat-Shock Proteins , Metabolism , Humans , Lactams, Macrocyclic , Pharmacology , Mechanistic Target of Rapamycin Complex 2 , Multiple Myeloma , Pathology , Multiprotein Complexes , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Signal Transduction , Sirolimus , Pharmacology , TOR Serine-Threonine Kinases , Metabolism
19.
Yonsei Medical Journal ; : 1497-1502, 2015.
Article in English | WPRIM | ID: wpr-177077

ABSTRACT

PURPOSE: Heat shock proteins (HSPs) are highly conserved molecular chaperones. There are various studies that assess the prognostic value of HSPs in patients with esophageal cancer, but the conclusion remains controversial. This is the first meta-analysis study aiming to summarize the evidence on the suitability of HSPs to predict patients' survival. MATERIALS AND METHODS: Searching PubMed, Web of science and Medline until May 31, 2014, data were compared for overall survival in patients with down-regulated HSPs level with those with up-regulated level. We conducted a meta-analysis of 9 studies (801 patients) that correlated HSPs levels with overall survival. Data were synthesized with hazard ratios (HRs). RESULTS: The estimated risk of death was 2.93-fold greater in HSP27 negative patients than HSP27 positive patients [95% confidence interval (CI), 1.12-7.62]. When limited to esophageal squamous cell carcinoma (ESCC), the risk of death in HSP27 negative patients seemed more significant (HR, 3.90; 95% CI, 2.35-6.49). Decreased expression of HSP70 was also associated with worse survival in esophageal cancer (HR, 2.83; 95% CI, 1.90-4.23) and, when limited to ESCC, HR was 3.21 (95% CI, 1.94-5.30). Data collected, however, were not sufficient to determine the prognostic value of HSP90 in patients with ESCC nor esophageal adenocarcinomas (EADC). CONCLUSION: In this meta-analysis, reduced HSP27 and HSP70 expressions were associated with poor survival in patients with esophageal cancer, especially esophageal squamous cell carcinoma.


Subject(s)
Adenocarcinoma/diagnosis , Carcinoma, Squamous Cell/diagnosis , Esophageal Neoplasms/diagnosis , Gene Expression Regulation, Neoplastic , HSP27 Heat-Shock Proteins , HSP70 Heat-Shock Proteins , HSP90 Heat-Shock Proteins , Heat-Shock Proteins/metabolism , Humans , Male , Neoplasm Proteins , Prognosis , Survival , Treatment Outcome
20.
Acta Pharmaceutica Sinica ; (12): 640-649, 2015.
Article in Chinese | WPRIM | ID: wpr-257088

ABSTRACT

HSP90, which is the biomarker of cell stress and endogenous protective protein, functions as a molecular chaperone. Many client proteins of HSP90, including EGFR, Met, Raf-1, IKK and p53, play important roles in the occurrence and development of tumor. Binding of HSP90 inhibitors triggers the deactivation of HSP90, resulting in client protein degradation, and hence inhibits the tumor growth by blocking multiple targets involved in signaling of tumor proliferation. This review summarizes recent development of small molecule inhibitors bound to N-terminal of HSP90.


Subject(s)
Antineoplastic Agents , Chemistry , HSP90 Heat-Shock Proteins , Humans , Neoplasms , Signal Transduction
SELECTION OF CITATIONS
SEARCH DETAIL