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1.
Biomed Res Int ; 2020: 5762932, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32309435

RESUMO

The magnetic field is the most common element in the universe, and high static magnetic field (HiSMF) has been reported to act as an inhibited factor for osteoclasts differentiation. Although many studies have indicated the negative role of HiSMF on osteoclastogenesis of RANKL-induced RAW264.7 cells, the molecular mechanism is still elusive. In this study, the HiSMF-retarded cycle and weakened differentiation of RAW264.7 cells was identified. Through RNA-seq analysis, RANKL-induced RAW264.7 cells under HiSMF were analysed, and a total number of 197 differentially expressed genes (DEGs) were discovered. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that regulators of cell cycle and cell division such as Bub1b, Rbl1, Ube2c, Kif11, and Nusap1 were highly expressed, and CtsK, the marker gene of osteoclastogenesis was downregulated in HiSMF group. In addition, pathways related to DNA replication, cell cycle, and metabolic pathways were significantly inhibited in the HiSMF group compared to the Control group. Collectively, this study describes the negative changes occurring throughout osteoclastogenesis under 16 T HiSMF treatment from the morphological and molecular perspectives. Our study provides information that may be utilized in improving magnetotherapy on bone disease.

2.
J Tissue Eng Regen Med ; 13(12): 2181-2190, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31622531

RESUMO

High static magnetic fields (HiSMFs) are usually defined as those SMFs with intensities ≥1 T. Although many studies have indicated that SMFs have positive effects on bone tissue, there were limited studies that investigate the effects of cells, including osteoclasts, to illustrate the effect of HiSMF on osteoclast differentiation, and whether iron involve in the altered osteoclast formation and resorption ability under HiSMF. 16 T HiSMF generated from a superconducting magnet was used. Osteoclastogenesis, bone resorption, acting ring formation, messenger ribonucleic acid expression, and protein expression were determined by tartrate-resistant acid phosphatase staining, pits formation assay, rhodamine-conjugated phalloidine staining, quantitative real-time polymerase chain reaction, and western blot, respectively. The changes induced by HiSMF in the level of iron and the concentration of mitochondrial protein, adenosine triphosphate, reactive oxygen species, malonaldehyde, and glutathione were examined by atomic absorption spectrometry and corresponding commercial kits, respectively. The results showed that HiSMF significantly inhibited osteoclastic formation and resorption ability and reduced cellular iron content during osteoclast differentiation. Mitochondrial concentration and oxidative stress levels in osteoclasts were decreased under HiSMF. Mechanistically, HiSMF markedly blocked the expression of osteoclast-associated transcription factors and osteoclast marker genes and inhibited iron absorption and iron storage-related protein expression. These findings demonstrated that the effect of HiSMF on iron metabolism of osteoclasts was involved in the inhibition of HiSMF on osteoclast differentiation.

3.
Cancers (Basel) ; 11(10)2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-31615107

RESUMO

One of many types of extracellular vesicles (EVs), exosomes are nanovesicle structures that are released by almost all living cells that can perform a wide range of critical biological functions. Exosomes play important roles in both normal and pathological conditions by regulating cell-cell communication in cancer, angiogenesis, cellular differentiation, osteogenesis, and inflammation. Exosomes are stable in vivo and they can regulate biological processes by transferring lipids, proteins, nucleic acids, and even entire signaling pathways through the circulation to cells at distal sites. Recent advances in the identification, production, and purification of exosomes have created opportunities to exploit these structures as novel drug delivery systems, modulators of cell signaling, mediators of antigen presentation, as well as biological targeting agents and diagnostic tools in cancer therapy. This review will examine the functions of immunocyte-derived exosomes and their roles in the immune response under physiological and pathological conditions. The use of immunocyte exosomes in immunotherapy and vaccine development is discussed.

4.
Mol Diagn Ther ; 23(5): 579-601, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31325035

RESUMO

Bladder cancer (BC) is the second highest morbid malignancy of the urinary tract and the fifth most common cancer worldwide. BC is highly malignant with significant morbidity and mortality, especially muscle-invasive BC (MIBC), which has a poor prognosis and frequently recurs after the first resection. Therefore, more sensitive diagnostic tools and effective therapeutic methods are urgently needed. MicroRNAs (miRNAs) are small noncoding RNAs that regulate the expression of protein-coding genes by repressing their translation or cleaving RNA transcripts in a sequence-specific manner. miRNAs play very important roles in regulating genes related to tumorigenesis, tumor development, progression, metastasis and angiogenesis. With the rapid development of high-throughput sequencing technology, an increasing number of miRNAs with aberrant expression between either BC patients and healthy volunteers or between BC tumor tissues and matched peripheral control tissues have been recently examined. The tumor etiopathogenesis must be determined to promote the development of new markers as diagnostic and prognostic tools and targets for bladder tumor therapy, it is therefore vital to elucidate the function of miRNAs with aberrant expression in BC. In the present study, we examined the published data of BC-related miRNAs by reviewing their expression levels, possible functions, potential target genes, related molecular regulatory networks, candidate markers for prognosis and diagnosis, and prospective therapeutic cases, and we summarized the status of research on BC-related miRNAs in recent years.


Assuntos
Biomarcadores Tumorais , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias da Bexiga Urinária/genética , Exossomos , Humanos , Técnicas de Diagnóstico Molecular , Gradação de Tumores , Estadiamento de Neoplasias , Prognóstico , Interferência de RNA , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/terapia
5.
Psychiatr Danub ; 31(1): 95-101, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30948695

RESUMO

BACKGROUND: Delirium, which is one of the most disturbing postoperative complications in elderly patients, shows high morbidity in patients undergoing lung cancer surgery. Dexmedetomidine (DEX) is considered a potential prophylactic agent for preventing patients' delirium after lung cancer surgery. SUBJECTS AND METHODS: Medical records of lung cancer patients over 65 years old with radical pulmonary resection at Henan Provincial People's Hospital from January 2015 to December 2017, China, were evaluated. Patients, care-providers, and investigators were all blinded to group assignment. DEX was administered in the preoperative and intraoperative periods. The incidence of delirium was calculated based on the Intensive Care Delirium Screening Checklist (ICDSC). Scores of ≥4 and 1-3 points represent the diagnoses of delirium and a pre-delirious state, respectively. RESULTS: During postoperative day 1 (POD 1) to POD 7, delirium occurs in both groups. During postoperative POD 1 to POD 7, the incidence of delirium is lower in the DEX group than that in the control group. Furthermore, there are more mild delirium patients but fewer moderate and severe delirium patients in the DEX group compared with the control group. Finally, patients in the DEX group have a shorter duration of delirium, lower numeric pain rating scale during movement and better sleep quality. CONCLUSION: Preoperative and intraoperative application of DEX can reduce the incidence and intensity of delirium after pulmonary resection in elderly patients with lung cancer.


Assuntos
Delírio , Dexmedetomidina , Neoplasias Pulmonares , Idoso , China , Delírio/prevenção & controle , Dexmedetomidina/uso terapêutico , Delírio do Despertar , Humanos , Hipnóticos e Sedativos/uso terapêutico , Neoplasias Pulmonares/cirurgia , Complicações Pós-Operatórias , Estudos Retrospectivos
6.
Adv Healthc Mater ; 8(10): e1900047, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30920772

RESUMO

Manganese dioxide (MnO2 )-based nanoparticles are a promising tumor microenvironment-responsive nanotheranostic carrier for targeted magnetic resonance imaging (MRI) and for alleviating tumor hypoxia. However, the complexity and potential toxicity of the present common synthesis methods limit their clinical application. Herein, multifunctional hyaluronic acid-MnO2 nanoparticles (HA-MnO2 NPs) are synthesized in a simple way by directly mixing sodium permanganate with HA aqueous solutions, which serve as both a reducing agent and a surface-coating material. The obtained HA-MnO2 NPs show an improved water-dispersibility, fine colloidal stability, low toxicity, and responsiveness to the tumor microenvironment (high H2 O2 and high glutathione, low pH). After intravenous injection, HA-MnO2 NPs exhibit a high imaging sensitivity for detecting rat intracranial glioma with MRI for a prolonged period of up to 3 d. These nanoparticles also effectively alleviate the tumor hypoxia in a rat model of intracranial glioma. The downregulation of VEGF and HIF-1α expression in intracranial glioma validates the sustained attenuation effect of HA-MnO2 NPs on tumor hypoxia. These results show that HA-MnO2 NPs can be used for sensitive, targeted MRI detection of gliomas and simultaneous attenuation of tumor hypoxia.

7.
Iran J Public Health ; 47(6): 803-813, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30087865

RESUMO

Background: Cancer is the leading cause of death in human disease and is a major public health problem around the world. Exosomes are a promising cancer biomarker and therapy target. Recent evidence demonstrate that tumor cells could inhibit natural killer (NK) cells' immune surveillance function by releasing exosomes into tumor microenvironment. The intercelluar uptake of tumor cell-derived exosomes by NK cells is vital for using these exosomes in tumor diagnose and therapy. We aimed to investigate the efficiency of NK cell uptake of tumor exosomes. Methods: Exosomes derived from different tumor cells, RAW264.7 cells and NK cells were labeled by fluorescent dye and co-cultured with NK cells. The uptake rates of NK cells were observed by fluorescence microscope and analyzed by flow cytometry. Results: NK cells could take up more exosomes from themselves and cell lines originating from bone marrow. Epithelial cell lines can take up more exosomes from epithelial cells. There was no significant difference in uptake efficiency between Jurkat cells and RAW264.7 cells by NK cells, indicating that maybe the origin other than species affects the efficiency of recipient cell uptake of exosomes. Different tumor cells derived exosomes had different uptake efficiency by NK cells. Conclusion: There is certain pattern of NK cells uptake tumor exosomes, which provide important insights on how tumors affect NK cells and develop appropriate countermeasures. In addition, it can be also helpful to select and design proper exosomes as a drug carrier in future.

8.
Exp Cell Res ; 363(2): 141-150, 2018 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-29269076

RESUMO

Extracellular vesicles (EVs) are nano vesicular structures that are secreted by almost all kinds of cells. Exosomes are small EVs derived from endosomes, with a diameter between 30-100nm. Tumour-derived exosomes carry many molecules and factors from tumour cells. These exosomes are recognized and taken up by immunocytes. However, tumour-derived exosomes can not only suppress immune cell functions but also help tumours escape immune surveillance in the tumour microenvironment. The present work investigated the effect of exosomes derived from genetical modified K562 cells (GMK cells), which express IL-15, IL-18 and 4-1BBL (TNFSF9) on their surface. The results showed that these GME exosomes, carrying IL-15, IL-18 and 4-1BBL proteins similar to their host cells, could activate NK cells, increase the cytotoxicity of NK cells on some tumour cells in a short treatment (4h) and promote NK cells proliferation. However, with an extended treatment time (48h), these exosomes could inhibite the cytotoxicity of NK cells by inhibiting activated receptor expression on NK cells. These results indicated the bifacial effects of GMK exosomes on NK cells, which will be helpful to explore the possibility of using transformed exosomes as an anti-tumour immune vaccine or a therapeutic tool in future.


Assuntos
Engenharia Celular , Citotoxicidade Imunológica/imunologia , Exossomos/metabolismo , Vesículas Extracelulares/metabolismo , Células Matadoras Naturais/imunologia , Engenharia Celular/métodos , Proliferação de Células/fisiologia , Humanos , Células K562 , Transdução de Sinais/fisiologia , Microambiente Tumoral/fisiologia
9.
Oncol Lett ; 13(3): 1539-1546, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28454288

RESUMO

Human aspartyl-(asparaginyl)-ß-hydroxylase (HAAH) has recently been the subject of several studies, as it was previously observed to be overexpressed in numerous types of carcinoma cells and tissues in patient tumor samples. HAAH has been implicated in tumor invasion and metastasis, indicating that it may be an important target and biomarker for tumor diagnosis and treatment. However, the immunological tools currently available for the study of this protein, including monoclonal antibodies, are limited, as is the present knowledge regarding the role of HAAH in tumor therapy and diagnosis. In the present study, a recombinant C-terminal domain of HAAH was expressed in Pichia pastoris and a novel monoclonal antibody (mAb) targeting HAAH (HAAH-C) was constructed. Immunofluorescence and antibody-dependent cellular cytotoxicity (ADCC) assays were used to demonstrate the specificity and ADCC activity of this antibody. The results demonstrated that this anti-C-terminal HAAH mAB, in combination with an existing anti-N terminal HAAH mAb, exhibited a high response to native HAAH from carcinoma cell culture supernatant, as measured with a double antibody sandwich enzyme-linked immunosorbent assay. This validated novel mAB-HAAH-C may prompt further studies into the underlying mechanisms of HAAH, and the exploration of its potential in tumor diagnosis and therapy.

10.
Pharm Biol ; 55(1): 799-809, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28112016

RESUMO

CONTEXT: Sonchus oleraceus L. (Asteraceae) (SO) is a dietary and traditional medicinal plant in China. However, its underlying mechanism of action as an anti-inflammatory agent is not known. OBJECTIVE: This study evaluates the anti-inflammatory activity of aqueous extract of SO. MATERIALS AND METHODS: The extract of SO was used to treat RAW 264.7 cells (in the working concentrations of 500, 250, 125, 62.5, 31.3 and 15.6 µg/mL) for 24 h. Pro-inflammatory cytokines and mediators produced in LPS-stimulated RAW 264.7 cells were assessed. Meanwhile, the expression level of TLR-4, COX-2, pSTATs and NF-κB was tested. Moreover, the anti-inflammatory activity of the extract in vivo was assessed using xylene-induced mouse ear oedema model and the anti-inflammatory compounds in the extracts were analyzed by HPLC-MS. RESULTS: SO extract significantly inhibited the production of pro-inflammatory cytokines and mediators at gene and protein levels with the concentration of 31.3 µg/mL, and suppressed the expression of TLR-4, COX-2, NF-κB and pSTAT in RAW 264.7 cells. The anti-inflammatory activity of SO in vivo has significant anti-inflammatory effects with the concentration of 250 and 125 mg/kg, and less side effect on the weights of the mice at the concentration of 250 mg/kg. Moreover, HPLC-MS analysis revealed that the anti-inflammatory compounds in the extract were identified as villosol, ferulaic acid, ß-sitosterol, ursolic acid and rutin. DISCUSSION AND CONCLUSION: This study indicated that SO extract has anti-inflammatory effects in vitro and in vivo, which will be further developed as novel pharmacological strategies in order to defeat inflammatory diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Lipopolissacarídeos/farmacologia , Extratos Vegetais/farmacologia , Sonchus , Animais , Sobrevivência Celular/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/farmacologia , Citocinas/antagonistas & inibidores , Citocinas/genética , Masculino , Camundongos , NF-kappa B/análise , Extratos Vegetais/análise , Células RAW 264.7 , Sonchus/química , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/genética
11.
J Ethnopharmacol ; 185: 289-99, 2016 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-27001625

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Sonchus oleraceus (L.) L (SO) and Juniperus sabina L (JS) are traditional medicinal plants in China. And the aqueous extracts of them have been used to treat tumor, inflammatory diseases, infection and so on in Chinese folk culture. However, the underlying mechanisms of their anti-tumor activities have not been illustrated yet. OBJECTIVE: This study aims to evaluate the inhibitory effects of aqueous extracts from SO and JS on tumor cells. MATERIALS AND METHODS: The prepared aqueous extracts of SO and JS were used to treat HepG-2 and K562 tumor cells, while the human peripheral blood mononuclear cells (PBMCs) were set as normal control. The viabilities, cell cycle and apoptosis of tumor cells after extracts treatment were assessed, in addition the expression of apoptosis-related genes (FasL, caspase 3, 6, 7, 8, 9, and 10) were analyzed. Meanwhile, the adherence and migration of HepG-2 were tested, and the expression levels of MMPs and ICAM-1 were analyzed. On top of that, the pSTAT in the two cells were also analyzed and suggested the related signaling pathway that the extracts acted on with in these tumor cells. RESULTS: Results showed that aqueous extracts of SO and JS have inhibitory effects on HepG-2 and K562 cells by decreasing cell viability and inducing apoptosis via up-regulation of the expression of the apoptosis-related genes FasL, caspase 3 and caspase 9. The extracts had different IC50 on tumor cells and PBMCs, which could block the tumor cell cycle at the G(0)/G(1) stage and significantly inhibit the adherence of HepG-2 cells. The extracts inhibited migration of these cells by inhibiting the expression of ICAM-1, MMP-2 and MMP-9. Further study indicated that the inhibition of pSTAT1 and 3 might be responsible for the inhibitory effects of the extracts on tumor cells. DISCUSSION AND CONCLUSION: The results of this study indicated that SO and JS extracts had the anti-tumor effects, which may be developed as novel anti-tumor drugs and used in cancer therapy.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Juniperus/química , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Sonchus/química , Antineoplásicos Fitogênicos/química , Apoptose , Adesão Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , China , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Temperatura Alta , Humanos , Células K562 , Fitoterapia , Extratos Vegetais/química , Regulação para Cima , Água
12.
Immunopharmacol Immunotoxicol ; 38(2): 77-86, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26642940

RESUMO

Heat shock protein 90 (Hsp90) is a ubiquitously expressed ATP-dependent molecular chaperone across all species that helps to the correct the folding of many proteins related to important signaling pathways. Tumor cells expressing Hsp90 have more ATP-binding affinity than normal cells. Many correlative inhibitors have been developed to promising anti-tumor strategies and have been evaluated in clinical trials. However, the effect of Hsp90 inhibitors on immunocytes cannot be ignored. Natural killer (NK) cells are key components of the innate immune system that play a pivotal role in tumor surveillance. The present study has investigated the potential effect of four Hsp90 inhibitors (NVP-AUY922, BIIB021, 17-DMAG, and SNX-2112) on human primary NK cells. The viability, cytotoxicity, apoptosis, phenotype, and cytokine secretion of NK cells after inhibitor treatment were assessed. The results of this study demonstrated that the inhibitors had negative effects on NK cell activity in a dose-dependent manner. The four inhibitors significantly reduced the cytotoxicity of the NK cells by decreasing viability, inducing apoptosis and down-regulating the expression of cytokines and functional receptors. These findings suggest that more attention should be given to the effect of Hsp90 inhibitors on NK cell function during clinical trials and also represent a potential immunosuppressant strategy.


Assuntos
Adenina/análogos & derivados , Benzoquinonas/farmacologia , Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Isoxazóis/farmacologia , Células Matadoras Naturais/metabolismo , Lactamas Macrocíclicas/farmacologia , Piridinas/farmacologia , Resorcinóis/farmacologia , Adenina/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Feminino , Humanos , Masculino
13.
Immunobiology ; 220(7): 876-88, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25758713

RESUMO

Natural killer (NK) cells (CD56(+)CD3(-)) are large, granular immunocytes that play a very pivotal role in the anti-inflammatory response and tumor surveillance. As an ideal cytotoxic lymphocyte (CTL), NK cells have attracted much attention in clinical trials. However, an insufficient number and their limited life span are bottlenecks that limit the application of NK cells in adoptive immunotherapy. Interleukins such as IL-2, IL-15 and IL-18 are recognized as factors that stimulate NK cells and have been used in NK cells ex vivo expansion. Similar to IL-2 and IL-15, IL-21 is a common γ-chain cytokine that is important in NK cell activation, maturation and proliferation. The present study aims to assess the effects of membrane-bound and soluble IL-21 on primary human NK cells during ex vivo expansion. IL-21 was found to have multiple effects on NK cells, increasing their cytotoxicity in a concentration-dependent manner by up-regulating IFN-γ and Granzyme-B expression. Nevertheless, at a high concentration (50 ng/mL), IL-21 curtailed the life span of NK cells by significantly inducing apoptosis. Moreover, when treated with IL-21, the number of NKT (CD56(+)CD3(+)) cells increased among peripheral blood mononuclear cells (PBMCs) during ex vivo expansion in a concentration-dependent manner. IL-21 also promoted expanded cells to enter into S phase of the cell cycle during the first to second weeks of culture. All these results suggest that IL-21 has multiple effects on NK cell development and functions. More attention should be given to the dosage and multiple effects of IL-21 when it was applied to NK cells in ex vivo expansion.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Citotoxicidade Imunológica/efeitos dos fármacos , Interleucinas/farmacologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Apoptose/imunologia , Diferenciação Celular/imunologia , Células Cultivadas , Granzimas/biossíntese , Humanos , Interferon gama/biossíntese , Interleucinas/imunologia , Células K562 , Contagem de Leucócitos , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos
14.
Iran J Public Health ; 44(12): 1632-42, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26811814

RESUMO

BACKGROUND: The humbug gene is a truncated isoform of Aspartyl ß-hydroxylase (ASPH) gene that is overexpressed in many human malignancies. In recent years, since humbug has received increasing attention, it is considered as a potential therapeutic molecular target. Therefore, it is necessary for preparing humbug protein and its monoclonal antibody to investigate its structure and function. METHOD: The optimized humbug gene, synthesized by Genscript in Nanjing, China on December 21st 2013, was expressed in Pichia pastoris cells that were cultured in a 10-L bioreactor. The recombinant protein was further obtained and purified by using ion exchange chromatography and Sephadex G75. The humbug protein was used to immunize Balb/c mice to generate the monoclonal antibodies. The specificity and sensitivity of the monoclonal antibodies were assessed by indirect enzyme-linked immunosorbent assay. Finally, the humbug monoclonal antibodies were used to detect the expression of humbug in several tumor cell lines via indirect immunofluorescence. RESULTS: Firstly, the recombinant humbug was expressed in P. pastoris successfully and efficiently by using a gene-optimized strategy. Secondly, the purification process of humbug was established via multiple chromatography methods. In addition, four monoclonal antibodies against humbug were obtained from the immunized Balb/c mice, and the result of indirect immunofluorescence was indicated that the humbug monoclonal antibody showed the high affinity with humbug protein, which expressed in several tumor cell lines. CONCLUSION: The over-expression of recombinant humbug provides adequate sources for its structural study and the preparation of the humbug-specific monoclonal antibody can potentially be used in tumor initial diagnosis and immunotherapy.

15.
J Agric Food Chem ; 62(45): 10928-35, 2014 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-25360711

RESUMO

Resveratrol (RES) is a polyphenol phytoalexin from plants, which has been reported to possess a variety of biological effects. The properties of RES on human natural killer (NK) cells were assessed in this study. Results showed that RES has concentration-dependent biphasic effects on NK cells. In high concentration (50 µM), RES can inhibit viability and promoted apoptosis of NK cells and human lymphoblastoid T (Jurkat) cells, which may affect the caspase signaling pathway. The Jurkat cells were more sensitive than NK cells on the RES caused cell death. However, when the concentration range reduced from 3.13 to 1.56 µM, RES showed the positive effects on NK cells by increasing the NK cells cytotoxicity via up-regulating the expression of NKG2D and IFN-γ (in mRNA and protein levels). These results indicated that one needs to pay more attention to the dosage and biphasic effects when RES was applied as antitumor drugs or health products.


Assuntos
Células Matadoras Naturais/efeitos dos fármacos , Estilbenos/análise , Estilbenos/farmacologia , Apoptose/efeitos dos fármacos , Humanos , Células Jurkat , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Resveratrol , Transdução de Sinais/efeitos dos fármacos
16.
Int Immunopharmacol ; 23(2): 452-9, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25281391

RESUMO

Natural killer (NK) cells are a key component of the innate immune system and play pivotal roles as inflammatory regulators and in tumor surveillance. Human aspartyl ß-hydroxylase (HAAH) is a plasma membrane and endoplasmic reticulum protein with hydroxylation activity, which is over-expressed in many malignant neoplasms and can be detected from the sera of tumor patients. HAAH is involved in regulating tumor cell infiltration and metastasis. Escaping from immune surveillance may help tumor cell infiltration and metastasis. However, the effects of HAAH on tumor immune surveillance have not yet been investigated carefully. The present study investigated the potential use of HAAH as an immune regulator of human NK cells. We assessed the effects of recombinant HAAH (r-HAAH) on primary human NK cell morphology, viability, cytotoxicity, apoptosis, receptors expression and cytokine/cytolytic proteins production. Our results demonstrated that r-HAAH negatively affects NK cell activity in a time and dose-dependent manner. It noticeably reduces the viability of the NK cells by increasing apoptosis and necrosis via caspase signaling pathways. Moreover, r-HAAH reduces the NK cell cytotoxicity by inhibiting surface expression of NKG2D, NKp44 and IFN-γ secretion. These findings suggest that one of the ways by which HAAH actively promotes tumor formation and proliferation is by inhibiting NK cell-surveillance activity.


Assuntos
Células Matadoras Naturais/citologia , Células Matadoras Naturais/fisiologia , Oxigenases de Função Mista/farmacologia , Linhagem Celular Tumoral , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Receptores de Células Matadoras Naturais/genética , Receptores de Células Matadoras Naturais/metabolismo , Proteínas Recombinantes
17.
Carbohydr Polym ; 101: 819-27, 2014 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-24299844

RESUMO

Polysaccharides are believed to be strong immunostimulants that can promote the proliferation and activity of T cells, B cells, macrophages and natural killer (NK) cells. This study aimed to investigate the effects of five polysaccharides (Grifola frondosa polysaccharide (GFP), lentinan (LNT), G. lucidum polysaccharide (GLP), Lycium barbarum polysaccharide (LBP) and yeast glucan (YG)) on primary human NK cells under normal or simulated microgravity (SMG) conditions. Our results demonstrated that polysaccharides markedly promoted the cytotoxicity of NK cells by enhancing IFN-γ and perforin secretion and increasing the expression of the activating receptor NKp30 under normal conditions. Meanwhile polysaccharides can enhance NK cell function under SMG conditions by restoring the expression of the activating receptor NKG2D and reducing the early apoptosis and late apoptosis/necrosis. Moreover, the antibody neutralization test showed that CR3 may be the critical receptor involved in polysaccharides induced NK cells activation. These findings indicated that polysaccharides may be used as immune regulators to promote the health of the public and astronauts during space missions.


Assuntos
Células Matadoras Naturais/citologia , Células Matadoras Naturais/efeitos dos fármacos , Polissacarídeos/farmacologia , Simulação de Ausência de Peso/efeitos adversos , Proliferação de Células/efeitos dos fármacos , Humanos , Células K562 , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Receptores de Complemento/metabolismo , Transdução de Sinais/efeitos dos fármacos
18.
Environ Toxicol Pharmacol ; 36(3): 948-55, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24035925

RESUMO

Natural killer (NK) cells play a pivotal role in human immunologic surveillance. Formaldehyde (FA), a ubiquitous environmental contaminant, has been classified as a carcinogen to humans. Although it is known that immune cells are sensitive to FA, so far little is known about how it's affecting the activity of human NK cells. To probe it, the primary human NK cells were treated with different concentrations of FA (3200, 1600, 800, 400, 200, 100, 50, and 0 µM) in vitro. The morphology, viability, apoptosis, cytotoxicity (killing tumor cell activity) and cytokine and cytolytic proteins secretion of NK cells were evaluated respectively. Our results reveal that FA could induce NK cells death obviously in a concentration-dependent manner. With the decreased concentrations of FA from 3200 µM to 800 µM, accordingly, the viability of NK cells increased from 65. 2 ± 12.1% to 78.48 ± 10.3% (p<0.05), and the cytotoxicity of NK cells recovered from 29.2 ± 8.5% to 63.9 ± 5.9% (p<0.05). The secretion of perforin was affected significantly by FA, whereas the secretion of IFN-γ and granzyme-B altered slightly. It is concluded that human NK cell is sensitive to FA, 800 µM may be a critical concentration of FA inhibiting the activity of human NK cell.


Assuntos
Poluentes Ambientais/toxicidade , Formaldeído/toxicidade , Células Matadoras Naturais/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Granzimas/metabolismo , Humanos , Interferon gama/metabolismo , Células Matadoras Naturais/ultraestrutura , Perforina/metabolismo
19.
Astrobiology ; 13(8): 703-14, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23919749

RESUMO

The deleterious effects of microgravity on lymphocytes have been demonstrated in previous studies. However, research on the effects of microgravity on human natural killer (NK) cells remains exceedingly limited. In this study, we demonstrated that NK cell cytotoxicity was significantly decreased under simulated microgravity (SMG) conditions (p<0.05). Several processes, including apoptosis, receptor expression, and cytokine secretion, were investigated in human NK cells under SMG. We observed decreased cytotoxicity, concurrent with increased apoptosis and necrosis, in NK cells after exposure to SMG (p<0.05). Additionally, interferon (IFN)-γ and perforin expression decreased significantly, and the expression of granzyme-B was only slightly reduced. Meanwhile, SMG selectively inhibited the expression of certain surface receptors on NK cells. Specifically, the expression of NKG2A and NKG2D were significantly downregulated under SMG, but the expression of NKp30 and NKp44 was not affected. We also found that interleukin (IL)-15 alone or in combination with IL-12 could counteract the inhibition of NK cell cytotoxicity under SMG. Our findings indicate that human NK cells were sensitive to SMG, as reflected by their decreased cytotoxicity. Factors such as increased early apoptosis and late apoptosis/necrosis and the decreased expression of INF-γ, cytolytic proteins, and cell surface receptors may be responsible for the loss of cytotoxicity in human NK cells under SMG. A combination of IL-12 and IL-15 may be useful as a therapeutic strategy for overcoming the effects of microgravity on human NK cells during long space missions.


Assuntos
Células Matadoras Naturais/fisiologia , Simulação de Ausência de Peso , Apoptose , Citocinas/metabolismo , Citometria de Fluxo , Humanos , Interleucinas , Células K562 , Reação em Cadeia da Polimerase em Tempo Real
20.
Sheng Wu Gong Cheng Xue Bao ; 27(4): 659-66, 2011 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-21848003

RESUMO

We investigated the mechanism of human aspartyl beta-hydroxylase (HAAH) in early diagnosis of tumors. The encoding gene of HAAH was cloned from the hepatic carcinoma by RT-PCR and expressed as a fused protein in the prokaryotic vector pBV-IL1. The expressed HAAH was purified by Ni(2+)-NTA purification column and the purified protein was then used to immunize Balb/c mice. Three hybridoma cell lines (respectively designated H3/E10, E4/F12 and G4/D8) stably expressing the monoclonal antibody specific to HAAH fusion protein were obtained. The specificity and sensitivity of the monoclonal antibody were assessed by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Finally, the monoclonal antibody expressed by H3/E10 cell line was used to detect the expression of HAAH in several tumor cell lines by indirect immuno-fluorescence, and the specific fluorescence was observed. In conclusion, this study successfully constructed the recombinant prokaryotic vector pBV-IL1-HAAH and prepared HAAH-specific monoclonal antibody for further study of the structure and function of the protein. The result may also lay solid foundation for the research of the molecular mechanism of HAAH in early diagnosis of tumors.


Assuntos
Anticorpos Monoclonais/biossíntese , Oxigenases de Função Mista/biossíntese , Oxigenases de Função Mista/imunologia , Proteínas Recombinantes/imunologia , Animais , Linhagem Celular Tumoral , Clonagem Molecular , Vetores Genéticos/genética , Humanos , Hibridomas/metabolismo , Imunização , Neoplasias Hepáticas/patologia , Camundongos , Camundongos Endogâmicos BALB C , Oxigenases de Função Mista/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética
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