Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 38
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Respir Res ; 21(1): 95, 2020 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-32321531

RESUMO

BACKGROUND: It is well established that airway remodeling and inflammation are characteristics for chronic obstructive pulmonary disease (COPD). Moreover, cigarette smoke extract (CSE) promots inflammation, apoptosis and oxidative stress in COPD. And, there is evidence suggested that alantolactone (ALT), a sesquiterpene lactone isolated from Inula helenium, plays an adverse role in inflammation, apoptosis and oxidative stress. However, few studies have investigated the function and mechanism of ALT treatment on the COPD pathological process. METHODS: The levels of IL-1 ß, TNF-α, IL-6 and IFN-γ were examined by ELISA. Cells' apoptosis and caspase-3 activity were detected by Cell Death Detection PLUS enzyme-linked immunosorbent assay and caspase-Glo 3/7 Assay, respectively. The content of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by using MDA and SOD assay kits. Reactive oxygen species (ROS) generation was measured by DCFH-DA assay. Protein expression was assayed by Western blot. RESULTS: In the present study, we aimed to observe the protective effects of ALT against inflammation, apoptosis and oxidative stress in human bronchial epithelial Beas-2B and NHBE cells. Our results showed that different doses of CSE exposure induced Beas-2B and NHBE cell inflammatory cytokines IL-1 ß, TNF-α, IL-6 and IFN-γ expression, cell apoptosis, caspase-3 activity and mediated oxidative stress markers MDA, ROS and SOD levels, while ALT treatment counteracted the effects of CSE. Further studies suggested that ALT attenuated NF-κB pathway activation. ALT also activated the Nrf2/HO-1 signal pathway through promoting Nrf2 nuclear aggregation and downstream HO-1 protein expression. HO-1 inhibitor tin protoporphyrin IX (SnPP IX) reversed the effects of ALT on Beas-2B and NHBE cell inflammation, apoptosis and oxidative stress. CONCLUSIONS: The above results collectively suggested that ALT suppressed CSE-induced inflammation, apoptosis and oxidative stress by modulating the NF-ĸB and Nrf2/ HO-1 axis.

2.
Int Immunopharmacol ; 83: 106475, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32283508

RESUMO

MicroRNAs (miRNAs) have emerged as critical modulators involved in the regulation of airway remodeling in asthma. MicroRNA-182-5p (miR-182-5p) has been reported as a key miRNA in regulating the proliferation and migration of various cell types, and its dysfunction contributes is implicated in a wide range of pathological processes. Yet, it remains unknown whether miR-182-5p modulates the proliferation and migration of airway smooth muscle (ASM) cells during asthma. In the present study, we aimed to determine the potential role of miR-182-5p in regulating the proliferation and migration of ASM cells induced by tumor necrosis factor (TNF)-α in vitro. We found that TNF-α stimulation markedly reduced miR-182-5p expression in ASM cells. Gain-of-function experiments showed that miR-182-5p upregulation suppressed the proliferation and migration of ASM cells induced by TNF-α. By contrast, miR-182-5p inhibition had the opposite effect. Notably, tripartite motif 8 (TRIM8) was identified as a target gene of miR-182-5p. TRIM8 expression was induced by TNF-α stimulation, and TRIM8 knockdown markedly impeded TNF-α-induced ASM cell proliferation and migration. Moreover, miR-182-5p overexpression or TRIM8 knockdown significantly downregulated the activation of nuclear factor-κB (NF-κB) induced by TNF-α. However, TRIM8 restoration partially reversed the miR-182-5p-mediated inhibitory effect on TNF-α-induced ASM cell proliferation and migration. In conclusion, our study indicates that miR-182-5p restricts TNF-α-induced ASM cell proliferation and migration through downregulation of NF-κB activation via targeting TRIM8. The results of our study highlight the potential importance of the miR-182-5p/TRIM8/NF-κB axis in the airway remodeling of asthma.

3.
Clin Exp Pharmacol Physiol ; 47(3): 449-458, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31665807

RESUMO

Deregulation of microRNAs (miRNAs) leads to malignant growth and aggressive invasion during cancer occurrence and progression. miR-147b has emerged as one of the cancer-related miRNAs that are dysregulated in multiple cancers. Yet, the relevance of miR-147b in non-small-cell lung cancer (NSCLC) remains unclear. In the present study, we aimed to report the biological function and signalling pathways mediated by miR-147b in NSCLC. Our results demonstrate that miR-147b expression is significantly downregulated in NSCLC tissues and cell lines. Overexpression of miR-147b decreased the proliferative ability, colony-forming capability, and invasive potential of NSCLC cells. Notably, our study identified ribosomal protein S15A (RPS15A), an oncogene in NSCLC, as a target gene of miR-147b. Our results showed that miR-147b negatively modulates RPS15A expression in NSCLC cells. An inverse correlation between miR-147b and RPS15A was evidenced in NSCLC specimens. Moreover, miR-147b overexpression downregulated the activation of Wnt/ß-catenin signalling via targeting of RPS15A. Overexpression of RPS15A partially reversed the miR-147b-mediated antitumour effect in NSCLC cells. Collectively, these findings reveal that miR-147b restricts the proliferation and invasion of NSCLC cells by inhibiting RPS15A-induced Wnt/ß-catenin signalling and suggest that the miR-147b/RPS15A/Wnt/ß-catenin axis is an important regulatory mechanism for malignant progression of NSCLC.

4.
Artif Cells Nanomed Biotechnol ; 47(1): 3621-3630, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31468992

RESUMO

With the arrival of the precision medicine and personalized treatment era, targeted therapy that improves efficacy and reduces side effects has become the mainstream approach of cancer treatment. Antibody fragments that further enhance penetration and retain the most critical antigen-specific binding functions are considered the focus of research targeting cancer imaging and therapy. Thanks to the superior penetration and rapid blood clearance of antibody fragments, antibody fragment-based imaging agents enable efficient and sensitive imaging of tumour sites. In tumour-targeted therapy, antibody fragments can directly inhibit tumour proliferation and growth, serve as an ideal carrier for delivery of anti-tumour drugs, or manipulate the immune system to eliminate tumour cells. In this review, the excellent physicochemical properties and the basic structure of antibody fragments are expressly depicted depicted, the progress of antibody fragments in cancer therapy and imaging are thoroughly summarized, and the future development of antibody fragments is predicted.


Assuntos
Diagnóstico por Imagem/métodos , Fragmentos de Imunoglobulinas/uso terapêutico , Terapia de Alvo Molecular/métodos , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Humanos , Fragmentos de Imunoglobulinas/imunologia , Neoplasias/imunologia
5.
Artif Cells Nanomed Biotechnol ; 47(1): 1476-1487, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31070063

RESUMO

Polymeric micelles (PMs) play a vital role in multidrug co-delivery and cancer treatment. However, the development of intelligent PMs further allows PMs to accurately -target tumour, selectively release cargo multidrug and increase uptake. Therefore, targets, controlled release and uptake of intelligent PMs should be paid more attention to improvement synergistic therapeutic outcomes and minimize side effects. In this review, tumour targeting of co-delivery intelligent PMs and its intracellular trafficking mechanisms were overviewed. And this review provides a comprehensive summarization of several intelligent co-delivery PMs. Such a system could control the multidrug to be released simultaneously or sequentially by special properties of tumour microenvironment (TME) (including acidic PH, redox, overexpressed enzyme, excessive temperature) and external environment trigger. Additionally, limitations, clinical translation and future perspectives of intelligent co-delivery PMs were also being discussed in this article.


Assuntos
Portadores de Fármacos/química , Micelas , Neoplasias/tratamento farmacológico , Polímeros/química , Humanos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Neoplasias/patologia , Microambiente Tumoral/efeitos dos fármacos
6.
ACS Appl Mater Interfaces ; 11(11): 10578-10588, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30802029

RESUMO

Hepatocellular carcinoma (HCC) poses a great threat to human health. The elegant combination of gene therapy and chemotherapy by nanocarriers has been repeatedly highlighted to realize enhanced therapeutic efficacy relative to monotreatment. However, the leading strategy to achieve the efficient codelivery of the gene and drug remains the electrostatic condensation with the nucleic acid and the hydrophobic encapsulation of drug molecules by the nanocarriers, which suffers substantially from premature drug leakage during circulation and severe off-target-associated side effects. To address these issues, we reported in this study the codelivery of liver-specific miRNA-122 and anti-cancer drug 5-fluorouracil (5-Fu) using a macromolecular prodrug approach, that is, electrostatic condensation with miRNA-122 using galactosylated-chitosan-5-fluorouracil (GC-FU). The delivery efficacy was evaluated comprehensively in vitro and in vivo. Specifically, the biocompatibility of GC-FU/miR-122 nanoparticles (NPs) was assessed by hemolysis activity analysis, BSA adsorption test, and cell viability assay in both normal liver cells (L02 cells) and endothelial cells. The resulting codelivery systems showed enhanced blood and salt stability, efficient proliferation inhibition of HCC cells, and further induction apoptosis of HCC cells, as well as downregulated expression of ADAM17 and Bcl-2. The strategy developed herein is thus a highly promising platform for an effective codelivery of miRNA-122 and 5-Fu with facile fabrication and great potential for the clinical translation toward HCC synergistic therapy.


Assuntos
Materiais Biocompatíveis/química , MicroRNAs/metabolismo , Pró-Fármacos/química , Proteína ADAM17/metabolismo , Animais , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Materiais Biocompatíveis/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Linhagem Celular , Quitosana/química , Regulação para Baixo/efeitos dos fármacos , Portadores de Fármacos/química , Sinergismo Farmacológico , Fluoruracila/química , Fluoruracila/farmacologia , Fluoruracila/uso terapêutico , Hemólise/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Camundongos , Camundongos Endogâmicos BALB C , MicroRNAs/química , Nanopartículas/química , Nanopartículas/toxicidade , Pró-Fármacos/farmacologia , Pró-Fármacos/uso terapêutico
7.
J Drug Target ; 27(4): 460-465, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30457021

RESUMO

Recurrence of lung adenocarcinoma has become one of the most frequent causes of major cancer incidence and mortality worldwide according to its frequently gained resistance to chemotherapies. In this study, we identified a poorly-studied kinase pyruvate dehydrogenase kinase isoform 2 (PDK2) as the most up-regulated kinase encoding gene in Cisplatin resistant lung adenocarcinoma. Additionally, PDK2-dependent Cisplatin-resistance promotes tumour growth of lung adenocarcinoma both in vitro and in vivo. Clinically, PDK2 expression was up-regulated in lung adenocarcinoma and was correlated to the poor prognosis of lung cancer patients. Mechanically, PDK2 promoted cell growth and Cisplatin-resistance of lung adenocarcinoma via transcriptional regulation of cyclin and CBS domain divalent metal cation transport mediator 3 (CNNM3), indicating that PDK2-CNNM3 signalling axis could be a potential therapeutic target for Cisplatin-resistant lung adenocarcinoma.

8.
Anticancer Agents Med Chem ; 18(14): 2062-2067, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30501603

RESUMO

BACKGROUND: Lung cancer is one of the most leading causes of cancer-related deaths in adults worldwide. Non-Small Cell Lung Cancer (NSCLC), which comprises 80 to 85% of all lung cancers, is the most lethal subtype of lung cancer with a 5-year survival of less than 13%. In this study, we identified a poorly-studied kinase PDK4 as the most up-regulated kinase encoding gene in Cisplatin resistant lung adenocarcinoma. METHODS: In vitro cell viability assay and in vivo tumor xenograft assay were used in the detection of cell proliferation. RNA isolation, quantitative Real-Time PCR, Western blot analysis, immunohistochemistry were used to investigate the expression of RNA and protein. Lentivirus infection was used to regulate gene expression. Luciferase assays were used to monitor EPAS1 promoter activity. RESULTS: In vivo PDK4 expression was elevated in a Cisplatin-resistant population of lung adenocarcinoma cells, PDK4-dependent Cisplatin-resistance promotes tumor growth of lung adenocarcinoma in vivo and in vitro, clinically PDK4 expression was associated with poor prognosis in lung adenocarcinoma patients, mechanically PDK4 promoted cell growth and Cisplatin-resistance of lung adenocarcinoma via transcriptional regulation of endothelial PAS domain-containing protein 1 (EPAS1). CONCLUSION: PDK4 is the most up-regulated kinase encoding gene in Cisplatin resistant lung adenocarcinoma and PDK4-dependent Cisplatin-resistance promotes tumor growth of lung adenocarcinoma mainly through transcriptional regulation of EPAS1. Enriched PDK4 expression was correlated with the poor prognosis of lung cancer patients, indicating that PDK4 could be a potential therapeutic target for Cisplatin-resistant lung adenocarcinoma.


Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/uso terapêutico , Cisplatino/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Proteínas de Homeodomínio/fisiologia , Neoplasias Pulmonares/tratamento farmacológico , Proteínas Proto-Oncogênicas/genética , Receptores Proteína Tirosina Quinases/genética , Ativação Transcricional/fisiologia , Adenocarcinoma/patologia , Animais , Linhagem Celular Tumoral , Células HEK293 , Humanos , Neoplasias Pulmonares/patologia , Camundongos , Camundongos Nus , Reação em Cadeia da Polimerase em Tempo Real
9.
Anticancer Agents Med Chem ; 18(12): 1680-1687, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30160218

RESUMO

BACKGROUND: Nrf2 pathway and autophagy are abnormally activated in response to cellular stress in various types of human cancers. In this study, we selected Beclin1 as an enter point to discuss the relationship between Nrf2 pathway and autophagy, and defined their associations with clinic pathological features and survival of the patients. METHOD: NSCLC specimens were processed for immunohistochemical and qRT-PCR to analyses the expression of Beclin1 and Nrf2. Kaplan-Meier method and log-rank test were used in the survival data. RESULTS: Beclin1 protein level was found to be significantly associated with more advanced TNM stage (P = 0.035), lymph node metastasis (P = 0.017) and distant metastasis (P = 0.005). The expression of Nrf2 protein was associated with larger tumor size (P = 0.032), more advanced TNM stage (P = 0.011), lymph node metastasis (P = 0.045) and distant metastasis (P = 0.013). Beside there was a strong inverse relationship between Beclin1 and Nrf2 expression in the NSCLC tissues. Distant metastasis, Beclin1, Nrf2, and Beclin1-/Nrf2+ expression was conformed to be independent prognostic factors of patients. CONCLUSION: Both Nrf2 overexpression and Beclin1 lower-expression are independent indicators of a poor prognosis in NSCLC patients.


Assuntos
Proteína Beclina-1/genética , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Fator 2 Relacionado a NF-E2/genética , Proteína Beclina-1/análise , Proteína Beclina-1/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Estimativa de Kaplan-Meier , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estrutura Molecular , Fator 2 Relacionado a NF-E2/análise , Fator 2 Relacionado a NF-E2/metabolismo , Relação Estrutura-Atividade , Análise de Sobrevida
10.
Inorg Chem ; 57(14): 8033-8036, 2018 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-29956541

RESUMO

The Brønsted basicity in activated metal-organic framework-808 (hereinafter denoted as MOF-808a) was confirmed by the analyses of CO2-TPD-MS, in situ DRIFTS, and acid-base titration. MOF-808a exhibited efficient recyclable catalytic activities for Heck coupling and oxidation of alcohol as a one-pot tandem reaction in base-free catalysis. It is the first evidence of the Brønsted basicity in zirconium metal-organic frameworks (Zr-MOFs) and gave rise to a new opportunity to extend the catalytic application of Zr-MOFs.

11.
J Investig Med ; 66(2): 334-339, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29141874

RESUMO

Pleural fibrosis can dramatically lower the quality of life. Numerous studies have reported that epithelial-mesenchymal transition (EMT) regulated by transforming growth factor-ß (TGF-ß) is involved in fibrosis. However, the molecular mechanism is inadequately understood. Fibroblast-specific protein-1 (S100A4) is a target of TGF-ß signaling. In our previous study, we have reported that S100A4 is highly expressed in pleural fibrosis. Thus, we suggest that S100A4 took part in the TGF-ß-induced EMT in pleural fibrosis. In this study, we determined the expression of S100A4 and EMT-related markers in Met-5A cells (pleural mesothelial cells) treated with TGF-ß or TGF-ß inhibitor by real-time PCR and western blot. In order to explore the role of S100A4, we used siRNA to knock down the expression of S100A4 in cell model. We found that the expression of epithelial cell marker was decreased and the mesenchymal cell marker increased with S100A4 upregulation after treatment with TGF-ß. Moreover, the changes of EMT-related event were restricted when the expression of S100A4 was knocked down. Conversely, S100A4 can partially rescue the EMT-related expression changes induced by TGF-ß inhibitor. These findings suggest that S100A4 expression is induced by the TGF-ß pathway, and silencing S100A4 expression can inhibit the process of TGF-ß-induced EMT.


Assuntos
Transição Epitelial-Mesenquimal , Epitélio/metabolismo , Pleura/citologia , Proteína A4 de Ligação a Cálcio da Família S100/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Biomarcadores/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Pirazóis/farmacologia , Quinolinas/farmacologia
12.
Lung Cancer ; 2017 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-28438350

RESUMO

This article has been withdrawn at the request of the Editor-in-Chief. Following peer-review and acceptance of the above referenced paper for publication in Lung Cancer, the Editor-in-Chief was contacted by the Editor-in-Chief of the journal, Gene Therapy, with information that the manuscript had simultaneously been submitted to both Lung Cancer and Gene Therapy. A referee selected to review the manuscript for Gene Therapy was also contacted by the Editor-in-Chief of the journal, Respiratory Research, with a request to review the same manuscript for that journal. The three journals ascertained that the manuscript had been simultaneously submitted to all three journals. In addition, as part of their investigation of potential simultaneous submission, the Editors of Lung Cancer compared the manuscript submitted to Gene Therapy with that accepted for publication in Lung Cancer, and this has raised concerns related to the data presented in the paper. The paper accepted for publication in Lung Cancer examines A549 and H810 cells. The paper submitted to Gene Therapy examines A549 and H510A cells. However, the data presented in both papers, including the figures, are identical. The Editors of Lung Cancer have asked the authors for an explanation, but the corresponding author has not responded. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/our-business/policies/article-withdrawal.

13.
Drug Deliv ; 24(1): 459-466, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28219253

RESUMO

A novel type of macromolecular prodrug delivery system is reported in this research. The N-galactosylated-chitosan-5-fluorouracil acetic acid conjugate (GC-FUA) based nanoparticle delivery system was evaluated in vitro and in vivo. Biocompatibility of GC-FUA-NPs was screened by BSA adsorption test and hemolysis activity examination in vitro. Cytotoxicity and cellular uptake study in HepG2 and A549 cells demonstrated that compared to free 5-Fu, the GC-FUA-NPs play great function in killing cancer cells for the cell endocytosis mediated by asialoglycoprotein receptor (ASGPR), which overexpresses on the cell surface. Pharmacokinetics study further illustrated that the drug-loaded nanoparticles has a much longer half-time than free 5-Fu in blood circulation in Sprague-Dawley (SD) rats. Tissue distribution was investigated in Kunming mice, and the result showed that the GC-FUA-NPs have a long circulation effect. The obtained data suggested that GC-FUA-NP is a very promising drug delivery system for efficient treatment of hepatocellular carcinoma.


Assuntos
Antimetabólitos Antineoplásicos/administração & dosagem , Carcinoma Hepatocelular/tratamento farmacológico , Quitosana/análogos & derivados , Quitosana/administração & dosagem , Portadores de Fármacos/administração & dosagem , Fluoruracila/análogos & derivados , Fluoruracila/administração & dosagem , Nanopartículas/química , Pró-Fármacos/administração & dosagem , Células A549 , Absorção Fisiológica , Animais , Antimetabólitos Antineoplásicos/efeitos adversos , Antimetabólitos Antineoplásicos/farmacocinética , Antimetabólitos Antineoplásicos/farmacologia , Carcinoma Hepatocelular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Quitosana/efeitos adversos , Quitosana/farmacocinética , Quitosana/farmacologia , Portadores de Fármacos/efeitos adversos , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Fluoruracila/efeitos adversos , Fluoruracila/farmacocinética , Fluoruracila/farmacologia , Glicosilação , Meia-Vida , Hemólise/efeitos dos fármacos , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/metabolismo , Camundongos , Nanopartículas/efeitos adversos , Nanopartículas/ultraestrutura , Pró-Fármacos/efeitos adversos , Pró-Fármacos/farmacocinética , Pró-Fármacos/farmacologia , Coelhos , Distribuição Aleatória , Ratos Sprague-Dawley , Distribuição Tecidual
14.
Cell Physiol Biochem ; 39(2): 685-92, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27442627

RESUMO

BACKGROUND/AIMS: The study was designed to explore the potential relationship of TLR4 and endothelial PAS domain-containing protein 1 (EPAS1) in vivo and vitro experiments. METHODS: Bronchoalveolar lavage fluid (BALF) samples were collected from 55 chronic obstructive pulmonary disease (COPD) patients and 25 healthy subjects. The differential cell count was performed using Wright-Giemsa staining. The expression levels of TLR4 and TLR5 were detected by RT-qPCR. The levels of methylation and mRNA expression of EPAS1 were assayed by bisulfite sequencing PCR and real-time PCR. The correlation of TLR4 and EPAS1 was also analyzed in TLR 4-overexpressing endothelial cells. RESULTS: The results showed that the number of neutrophils, lymphocytes and macrophages and expression of TLR 4 were significantly increased in lower respiratory tract of COPD patients. Moreover, decreased EPAS1 mRNA and increased EPAS1 promoter methylation were detected in COPD, which were closely associated with increased TLR4 expression. According to in vitro experiments, TLR 4 inhibited EPAS1 mRNA expression and promoted promoter methylation in endothelial cells. CONCLUSION: These findings suggest that TLR4 over-expression decreased EPAS1expression which contributes to the progress of COPD.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regulação da Expressão Gênica , Doença Pulmonar Obstrutiva Crônica/genética , Sistema Respiratório/metabolismo , Receptor 4 Toll-Like/genética , Western Blotting , Contagem de Células , Metilação de DNA , Humanos , Linfócitos/metabolismo , Macrófagos/metabolismo , Pessoa de Meia-Idade , Neutrófilos/metabolismo , Regiões Promotoras Genéticas/genética , Doença Pulmonar Obstrutiva Crônica/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Receptor 5 Toll-Like/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
15.
Curr Pharm Des ; 22(4): 506-13, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26517529

RESUMO

The development of slow release nano-sized carriers for efficient antineoplastic drug delivery with a biocompatible and biodegradable pectin-based macromolecular pro-drug for tumor therapy has been reported in this study. Pectin-doxorubicin conjugates (PDC), a macromolecular pro-drug, were prepared via an amide condensation reaction, and a novel amphiphilic core-shell micell based on a PDC macromolecular pro-drug (PDC-M) was self-assembled in situ, with pectin as the hydrophilic shell and doxorubicin (DOX) as the hydrophobic core. Then the chemical structure of the PDC macromolecular pro-drug was identified by both Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy ((1)H-NMR), and proved that doxorubicin combined well with the pectin and formed macromolecular pro-drug. The PDC-M were observed to have an unregularly spherical shape and were uniform in size by scanning electron microscopy (SEM). The average particle size of PDC-M, further measured by a Zetasizer nanoparticle analyzer (Nano ZS, Malvern Instruments), was about 140 nm. The encapsulation efficiency and drug loading were 57.82% ± 3.7% (n = 3) and 23.852% ±2.3% (n = 3), respectively. The in vitro drug release behaviors of the resulting PDC-M were studied in a simulated tumor environment (pH 5.0), blood (pH 7.4) and a lysosome media (pH 6.8), and showed a prolonged slow release profile. Assays for antiproliferative effects and flow cytometry of the resulting PDC-M in HepG2 cell lines demonstrated greater properties of delayed and slow release as compared to free DOX. A cell viability study against endothelial cells further revealed that the resulting PDC-M possesses excellent cell compatibilities and low cytotoxicities in comparison with that of the free DOX. Hemolysis activity was investigated in rabbits, and the results also demonstrated that the PDC-M has greater compatibility in comparison with free DOX. This shows that the resulting PDC-M can ameliorate the hydrophobicity of free DOX. This work proposes a novel strategy for in-situ one-step synthesis of macromolecular pro-drugs and fabrication of a core-shell micelle, demonstrating great potential for cancer chemotherapy.


Assuntos
Doxorrubicina/síntese química , Portadores de Fármacos/síntese química , Sistemas de Liberação de Medicamentos/métodos , Micelas , Pectinas/síntese química , Pró-Fármacos/síntese química , Animais , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Química Farmacêutica , Relação Dose-Resposta a Droga , Doxorrubicina/farmacologia , Portadores de Fármacos/farmacologia , Feminino , Células Hep G2 , Humanos , Substâncias Macromoleculares/síntese química , Masculino , Nanopartículas , Pectinas/farmacologia , Pró-Fármacos/farmacologia , Coelhos
16.
Inhal Toxicol ; 27(14): 822-31, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26671198

RESUMO

OBJECTIVE: Cigarette smoking is the main cause of chronic obstructive pulmonary disease (COPD) and may modulate the immune response of exposed individuals. Mast cell function can be altered by cigarette smoking, but the role of smoking in COPD remains poorly understood. The current study aimed to explore the role of cigarette smoke extract (CSE)-treated mast cells in COPD pathogenesis. METHODS: Cytokine and chemokine expression as well as degranulation of bone marrow-derived mast cells (BMMCs) were detected in cells exposed to immunoglobulin E (IgE) and various doses of CSE. Adoptive transfer of CSE-treated BMMCs into C57BL/6J mice was performed, and macrophage infiltration and polarization were evaluated by fluorescence-activated cell sorting (FACS). Furthermore, a coculture system of BMMCs and macrophages was established to examine macrophage phenotype transition. The role of protease serine member S31 (Prss31) was also investigated in the co-culture system and in COPD mice. RESULTS: CSE exposure suppressed cytokine expression and degranulation in BMMCs, but promoted the expressions of chemokines and Prss31. Adoptive transfer of CSE-treated BMMCs induced macrophage infiltration and M2 polarization in the mouse lung. Moreover, CSE-treated BMMCs triggered macrophage M2 polarization via Prss31 secretion. Recombinant Prss31 was shown to activate interleukin (IL)-13/IL-13Rα/Signal transducers and activators of transcription (Stat) 6 signaling in macrophages. Additionally, a positive correlation was found between Prss31 expression and the number of M2 macrophages in COPD mice. CONCLUSION: In conclusion, CSE-treated mast cells may induce macrophage infiltration and M2 polarization via Prss31 expression, and potentially contribute to COPD progression.


Assuntos
Misturas Complexas/toxicidade , Macrófagos Alveolares/fisiologia , Mastócitos/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/induzido quimicamente , Fumaça/efeitos adversos , Produtos do Tabaco/efeitos adversos , Animais , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Doença Pulmonar Obstrutiva Crônica/patologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Fumaça/análise , Produtos do Tabaco/análise , Triptases/genética , Triptases/metabolismo
18.
Mol Med Rep ; 11(4): 2555-61, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25482186

RESUMO

The aim of the present study was to investigate the feasibility of lentiviral­mediated Cys­Asn­Gly­Arg­Cys (CNGRC) peptide gene transduction in adipose stem cells. Adipose stem cells were prepared using enzymatic digestion and repeated adherence methods and identified in culture by immunofluorescence staining of surface markers. The pluripotency of the cultured adipose stem cells was confirmed by their induced differentiation into bone and fat cells. Following polymerase chain reaction amplification, the gene sequence for the CNGRC peptide was cloned into a lentiviral vector, which was then co­transfected into 293T cells with packaging plasmids Helper 1.0 and Helper 2.0. The lentiviruses carrying the CNGRC peptide gene were then harvested and used to transfect adipose stem cells. Following transduction, expression of CNGRC in adipose stem cells was detected using western blot analysis. Adipose stem cells in culture were successfully induced to differentiate into adipocytes and osteoblasts and the lentiviral vector containing CNGRC­3Flag­EGFP was successfully constructed. Following transduction, western blot analysis and immunofluorescence staining demonstrated expression of the CNGRC protein in adipose stem cells. This suggested that adipose stem cell lines expressing the CNGRC peptide were successfully established.


Assuntos
Tecido Adiposo/citologia , Vetores Genéticos/genética , Lentivirus/genética , Peptídeos Cíclicos/genética , Células-Tronco/metabolismo , Transdução Genética , Adipócitos/citologia , Animais , Antígenos de Superfície/metabolismo , Técnicas de Cultura de Células , Diferenciação Celular , Expressão Gênica , Imunofenotipagem , Masculino , Osteócitos/citologia , Fenótipo , Ratos , Células-Tronco/citologia
19.
Int J Oncol ; 46(3): 1141-8, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25544568

RESUMO

Breast cancer is the most common malignancy in women. The Notch signaling pathway has been shown to be associated with the development and progression of many human cancers, including breast cancer, but the precise mechanism remains unknown. Here, the influence of Notch1 signaling in mammary epithelial cells was studied. We showed that Notch1 promotes proliferation in MCF7 and MCF10A cells. Transwell assay indicated that Notch1 overexpression promotes cell migration and the invasion of breast cancer cells. We showed that MCF7 and MCF10A cells overexpressing Notch1 acquired features of epithelial-mesenchymal transition (EMT) and displayed a cancer stem cell (CSC) phenotype. The expression levels of the epithelial markers E-cadherin and occludin were decreased, while the expression levels of the mesenchymal markers N-cadherin, vimentin and fibronectin were increased in cells overexpressing Notch1. We demonstrated that Notch1 induced phosphorylation of the signal transducer and activator of transcription 3 (STAT3) in breast cancer cells and increased the expression of p65 and interleukin (IL)-1ß. Inhibition of STAT3 activity by JSI124 reduced the expression of p65 and IL-1. Treatment of MCF7-notch1 and MCF10A-notch1 cells with JSI124 also reduced the expression of N-cadherin, markers of epithelial mesenchymal transition and increased the expression of E-cadherin. Our results suggest that Notch1 promotes EMT and the CSC phenotype through induction of STAT3.


Assuntos
Neoplasias da Mama/patologia , Transição Epitelial-Mesenquimal/genética , Células-Tronco Neoplásicas/patologia , Receptor Notch1/fisiologia , Fator de Transcrição STAT3/fisiologia , Animais , Neoplasias da Mama/genética , Transformação Celular Neoplásica/genética , Células Cultivadas , Feminino , Humanos , Células MCF-7 , Camundongos , Camundongos Nus , Fenótipo , Transfecção
20.
Oncol Rep ; 31(5): 2358-64, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24676336

RESUMO

ARHI is a maternally imprinted tumor suppressor gene that is expressed in normal breast epithelial cells but not in most breast cancer cells. Aberrant methylation and hypernomic histone deacetylation have been implicated in the silencing of ARHI. To investigate the mechanism of ARHI induction, MDA-MB-231 breast cancer cells were either transfected with the eukaryotic expression vector, pcDNA3.1(+)-ARHI, or were simultaneously treated with a histone deacetylase inhibitor, [trichostatin A, (TSA)] and the methyltransferase inhibitor, 5-aza-2'-deoxycytidine (DAC). The latter treatment group also included the targeting of ARHI by small interfering RNA (siRNA) to further examine interactions between ARHI and the drugs applied. Levels of ARHI were detected by western blotting, MTT assays were used to evaluate cell proliferation, and both cell cycle progression and apoptosis were detected using flow cytometry. Both the transfection of pcDNA3.1(+)­ARHI and the application of TSA+DAC induced the expression of ARHI. Furthermore, reduced cell proliferation, cell cycle arrest and enhanced apoptosis were observed for both groups compared to controls. However, a G1/S cell cycle arrest was observed for the pcDNA3.1(+)-ARHI group, while a G2 cell cycle arrest was observed for the TSA+DAC group. The latter effect was reversed with the introduction of ARHI-targeted siRNA in combination with TSA+DAC treatment. To further clarify these observations, expression levels of several key cell cycle regulators were analyzed by western blotting. The pcDNA3.1(+)-ARHI group exhibited higher expression levels of p53, p21 and p27, and lower levels of cyclin D1, CDK4 and CDK6 when compared to the control group (P<0.05). For the TSA+DAC group, higher levels of p53, p21, cyclin B1 and Chk1 were detected, concomitant with lower levels of CDK1, when compared to the control group. Taken together, these results suggest that ARHI acts as a tumor suppressor gene in MDA-MB-231 cells and, although TSA+DAC can block the cells at different cell cycle phage, the antitumor effect is ARHI-dependent.


Assuntos
Azacitidina/análogos & derivados , Neoplasias da Mama/genética , Ácidos Hidroxâmicos/farmacologia , Proteínas rho de Ligação ao GTP/genética , Apoptose/efeitos dos fármacos , Apoptose/genética , Azacitidina/farmacologia , Neoplasias da Mama/tratamento farmacológico , Proteína Quinase CDC2 , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Quinase 1 do Ponto de Checagem , Ciclina B1/biossíntese , Ciclina D1/biossíntese , Quinase 4 Dependente de Ciclina/biossíntese , Quinase 6 Dependente de Ciclina/biossíntese , Inibidor de Quinase Dependente de Ciclina p21/biossíntese , Inibidor de Quinase Dependente de Ciclina p27/biossíntese , Quinases Ciclina-Dependentes/biossíntese , Decitabina , Feminino , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/genética , Vetores Genéticos/genética , Inibidores de Histona Desacetilases/farmacologia , Humanos , Metiltransferases/antagonistas & inibidores , Proteínas Quinases/biossíntese , Interferência de RNA , RNA Interferente Pequeno , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase S do Ciclo Celular/genética , Transfecção , Proteína Supressora de Tumor p53/biossíntese , Proteínas rho de Ligação ao GTP/biossíntese
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA