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1.
Braz Dent J ; 31(3): 304-309, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32667511

RESUMO

Among other factors, types of bisphosphonates and treatment regimens seem to be strongly associated with the success or failure of installation of osseointegrated implants. This study investigated the influence of two bisphosphonates, sodium alendronate (SA) and zoledronic acid (ZA), on the metabolism of osteoblasts. Human osteoblasts (Saos-2) were seeded onto machined or acid-treated titanium discs previously placed on 24-well plates in complete culture medium. After 24 h, cells were exposed to bisphosphonates at 0.5, 1 or 5 µM for 24 h, 48 h or 7 days. The effects of SA and ZA on osteoblasts were assessed based on the adhesion of these cells to the titanium surfaces by direct fluorescence, cell viability, total protein and collagen synthesis. Alkaline phosphatase activity and mineral nodule deposition by these cells were also evaluated. Data were evaluated by ANOVA and Tukey tests (α=0.05). Decreased adhesion of cells to the titanium discs was observed when exposed to both bisphosphonates; however, this lack of cell adhesion was more evident for ZA-treated cells. In addition, the exposure of osteoblasts to ZA decreased the viability, ALP activity and mineral nodule deposition, which may be related to poor osseointegration after implant installation.


Assuntos
Difosfonatos , Titânio , Fosfatase Alcalina , Adesão Celular , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Osteoblastos , Propriedades de Superfície , Ácido Zoledrônico
2.
Int J Nanomedicine ; 15: 2633-2646, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32368045

RESUMO

Objective: The aim of this study is to fabricate functional scaffolds to gene delivery bone morphogenetic protein-2 (BMP-2) plasmid for bone formation in bone tissue engineering. Methods: Dendriplexes (DPs) of generation 4 polyamidoamin (G4-PAMAM)/BMP-2 plasmid were prepared through microfluidic (MF) platform. The physiochemical properties and toxicity of DPs were evaluated by DLS, AFM, FESEM and MTT assay. In order to create a suitable environment for stem cell growth and differentiation, poly-l-lactic acid (PLLA) and poly-l-lactic acid/poly (ethylene oxide) (PLLA/PEO) scaffolds containing hydroxyapatite nanoparticles (HA) and DPs were fabricated by the electrospinning method. The osteogenic potency of the scaffolds on human adipose tissue-derived mesenchymal stem cells (hASCs) was investigated. Results: The results revealed that tuning the physical properties of DPs by adjusting flow parameters in microfluidic platform can easily improve the cell viability compared to conventional bulk mixing method. Also, the result showed that the presence of HA and DPs in PLLA/PEO scaffold enhanced alkaline phosphatase (ALP) activity and increased the amount of deposited Ca, as well as, related to osteogenesis gen markers. Conclusion: This study indicated that on using the MF platform in preparation of DPs and loading them along with HA in PLLA/PEO scaffold, the osteogenic differentiation of hASCs could be tuned.


Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Osso e Ossos/fisiologia , Durapatita/química , Microfluídica , Nanofibras/química , Poliaminas/química , Engenharia Tecidual/métodos , Tecidos Suporte/química , Fosfatase Alcalina/metabolismo , Cálcio/metabolismo , Adesão Celular , Morte Celular , Diferenciação Celular , Proliferação de Células , Forma Celular , DNA/metabolismo , Dendrímeros/química , Humanos , Células-Tronco Mesenquimais/metabolismo , Nanopartículas/química , Tamanho da Partícula , Plasmídeos/metabolismo , Poliésteres/química , Resistência à Tração
3.
PLoS One ; 15(4): e0232321, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32353084

RESUMO

Decorin is a member of small leucine-rich proteoglycan family, which is involved in multiple biological functions mainly as a structural and signaling molecule, and disturbances in its own metabolism plays a crucial role in the pathogenesis of osteoarthropathy. In this study, we aim to further explore the biological function of decorin and their role in human chondrocyte cell line, C28/I2. Lentivirus-mediated shRNA was applied to down-regulate decorin expression in C28/I2 chondrocytes. Effect of decorin knockdown on gene expression profiles was determined by RNA sequencing followed by bioinformatics analysis. MTT, adhesion assays and flow cytometry were used to investigate the effect of decorin knockdown on cell proliferation, adhesion, and apoptosis. sGAG content in the culture medium was determined by DMMB assay. Stably transfected C28/I2 cells were seeded onto the cancellous bone matrix gelatin (BMG) to construct tissue-engineered cartilage. The histological patterns were evaluated by H&E and Toluidine blue staining. In this study, 1780 differentially expressed genes (DEGs) including 864 up-regulated and 916 down-regulated genes were identified using RNA-Seq. The reliability of the gene expression was further verified by qRT-PCR. GO and KEGG pathway enrichment analysis revealed diverse cellular processes were affected by decorin silencing such as: cell adhesion, growth, and metabolism of extracellular matrix. In addition, we confirmed that down-regulation of decorin significantly suppressed cell proliferation and adhesion and induced apoptosis. The sGAG content in the media was significantly increased after decorin silencing. Engineered articular tissues in the decorin knockdown group exhibited cartilage destruction and proteoglycan loss as evidenced by H&E and Toluidine blue stains. Overall, this combined data helps to provide a comprehensive understanding of the roles of decorin following its knockdown in C28/I2 cells.


Assuntos
Adesão Celular , Proliferação de Células , Condrócitos/metabolismo , Decorina/metabolismo , Matriz Extracelular/metabolismo , Transcriptoma , Animais , Apoptose , Condrócitos/citologia , Condrócitos/fisiologia , Decorina/genética , Células HEK293 , Humanos , Coelhos
4.
PLoS One ; 15(5): e0232432, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32365067

RESUMO

CR3 and CR4, the leukocyte specific ß2-integrins, involved in cellular adherence, migration and phagocytosis, are often assumed to have similar functions. Previously however, we proved that under physiological conditions CR4 is dominant in the adhesion to fibrinogen of human monocyte-derived macrophages (MDMs) and dendritic cells (MDDCs). Here, using inflammatory conditions, we provide further evidence that the expression and function of CR3 and CR4 are not identical in these cell types. We found that LPS treatment changes their expression differently on MDMs and MDDCs, suggesting a cell type specific regulation. Using mAb24, specific for the high affinity conformation of CD18, we proved that the activation and recycling of ß2-integrins is significantly enhanced upon LPS treatment. Adherence to fibrinogen was assessed by two fundamentally different approaches: a classical adhesion assay and a computer-controlled micropipette, capable of measuring adhesion strength. While both receptors participated in adhesion, we demonstrated that CR4 exerts a dominant role in the strong attachment of MDDCs. Studying the formation of podosomes we found that MDMs retain podosome formation after LPS activation, whereas MDDCs lose this ability, resulting in a significantly reduced adhesion force and an altered cellular distribution of CR3 and CR4. Our results suggest that inflammatory conditions reshape differentially the expression and role of CR3 and CR4 in macrophages and dendritic cells.


Assuntos
Células Dendríticas/imunologia , Inflamação/imunologia , Integrina alfaXbeta2/imunologia , Antígeno de Macrófago 1/imunologia , Macrófagos/imunologia , Podossomos/imunologia , Anticorpos Bloqueadores/imunologia , Antígenos CD18/imunologia , Adesão Celular/imunologia , Adesão Celular/fisiologia , Diferenciação Celular/imunologia , Movimento Celular/imunologia , Movimento Celular/fisiologia , Células Dendríticas/patologia , Células Dendríticas/fisiologia , Fibrinogênio/imunologia , Humanos , Técnicas In Vitro , Inflamação/patologia , Lipopolissacarídeos/imunologia , Macrófagos/patologia , Macrófagos/fisiologia , Fagocitose/imunologia , Fagocitose/fisiologia , Podossomos/patologia
5.
Life Sci ; 254: 117780, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32407844

RESUMO

AIMS: In vivo studies suggest a positive influence of fresh frozen plasma (FFP) on endothelial properties and vascular barrier function, leading to improved outcomes in animal sepsis models as well as in major abdominal surgery. However, those effects are incompletely described. It was our aim to evaluate in vitro effects of FFP on endothelial key functions and to identify underlying mechanisms. MATERIALS AND METHODS: Human pulmonary microvascular endothelial cells (HPMECs) were prestimulated with LPS, followed by incubation with FFP. Permeability for FITC-dextran was assessed, and intercellular gap formation was visualized. NF-κB nuclear translocation and expression of pro-inflammatory, pro-adhesion, and leakage-related genes were evaluated, and monocyte adhesion to ECs was assessed. Intracellular cAMP levels as well as phosphorylation of functional proteins were analyzed. In patients undergoing major abdominal surgery, Syndecan-1 serum levels were assessed prior to and following FFP transfusion. KEY FINDINGS: Post-incubation of HPMVECs with FFP increased intracellular cAMP levels that had been decreased by preceding LPS stimulation. On one hand, this reduced endotoxin-mediated upregulation of IL-8, ICAM-1, VCAM-1, VEGF, and ANG-2. Impaired phosphorylation of functional proteins was restored, and intercellular cohesion and barrier function were rescued. On the other hand, NF-κB nuclear translocation as well as monocyte adhesion was markedly increased by the combination of LPS and FFP. Syndecan-1 serum levels were lower in surgery patients that were transfused with FFP compared to those that were not. SIGNIFICANCE: Our data provide evidence for a differential modulation of crucial endothelial properties by FFP, potentially mediated by elevation of intracellular cAMP levels.


Assuntos
Células Endoteliais/fisiologia , Endotélio Vascular/metabolismo , Plasma/fisiologia , Idoso , Adesão Celular/fisiologia , Permeabilidade da Membrana Celular/fisiologia , Células Cultivadas , AMP Cíclico/metabolismo , Dextranos/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Junções Comunicantes/fisiologia , Humanos , Lipopolissacarídeos , Pessoa de Meia-Idade , Monócitos/fisiologia , NF-kappa B/metabolismo , Fosforilação , Sindecana-1/sangue
6.
Invest Ophthalmol Vis Sci ; 61(3): 44, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32232343

RESUMO

Purpose: To determine the composition of extracellular matrix (ECM) proteins secreted by a conjunctival epithelial cell line and to identify components that aid conjunctival epithelial cell culture. Methods: Human conjunctival epithelial cell line (HCjE-Gi) cells were cultured in serum-free media and their ECM isolated using ammonium hydroxide. Growth characteristics were evaluated for fresh HCjE-Gi cells plated onto ECMs obtained from 3- to 28-day cell cultures. Mass spectrometry was used to characterize the ECM composition over 42 culture days. Cell adhesion and growth on pre-adsorbed fibronectin and α-2-HS-glycoprotein (α-2-HS-GP) were investigated. Results: Day 3 ECM provided the best substrate for cell growth compared to ECM obtained from 5- to 28-day cell cultures. Mass spectrometry identified a predominantly laminin 332 matrix throughout the time course, with progressive changes to matrix composition over time: proportional decreases in matrix-bound growth factors and increases in proteases. Fibronectin and α-2-HS-GP were 5- and 200-fold enriched as a proportion of the early ECM relative to the late ECM, respectively. Experiments on these proteins in isolation demonstrated that fibronectin supported rapid cell adhesion, whereas fibronectin and α-2-HS-GP both supported enhanced cell growth compared to tissue culture polystyrene. Conclusions: These data reveal α-2-HS-GP as a candidate protein to enhance the growth of conjunctival epithelial cells and raise the possibility of exploiting these findings for targeted improvement to synthetic tissue engineered conjunctival substrates.


Assuntos
Túnica Conjuntiva/metabolismo , Proteínas da Matriz Extracelular/metabolismo , alfa-2-Glicoproteína-HS/metabolismo , Biomarcadores/metabolismo , Adesão Celular/fisiologia , Contagem de Células , Linhagem Celular , Proliferação de Células/fisiologia , Túnica Conjuntiva/citologia , Meios de Cultura Livres de Soro , Células Epiteliais/metabolismo , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Humanos , Espectrometria de Massas
7.
Proc Natl Acad Sci U S A ; 117(16): 9064-9073, 2020 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-32273388

RESUMO

The invasive behavior of glioblastoma is essential to its aggressive potential. Here, we show that pleckstrin homology domain interacting protein (PHIP), acting through effects on the force transduction layer of the focal adhesion complex, drives glioblastoma motility and invasion. Immunofluorescence analysis localized PHIP to the leading edge of glioblastoma cells, together with several focal adhesion proteins: vinculin (VCL), talin 1 (TLN1), integrin beta 1 (ITGB1), as well as phosphorylated forms of paxillin (pPXN) and focal adhesion kinase (pFAK). Confocal microscopy specifically localized PHIP to the force transduction layer, together with TLN1 and VCL. Immunoprecipitation revealed a physical interaction between PHIP and VCL. Targeted suppression of PHIP resulted in significant down-regulation of these focal adhesion proteins, along with zyxin (ZYX), and produced profoundly disorganized stress fibers. Live-cell imaging of glioblastoma cells overexpressing a ZYX-GFP construct demonstrated a role for PHIP in regulating focal adhesion dynamics. PHIP silencing significantly suppressed the migratory and invasive capacity of glioblastoma cells, partially restored following TLN1 or ZYX cDNA overexpression. PHIP knockdown produced substantial suppression of tumor growth upon intracranial implantation, as well as significantly reduced microvessel density and secreted VEGF levels. PHIP copy number was elevated in the classical glioblastoma subtype and correlated with elevated EGFR levels. These results demonstrate PHIP's role in regulating the actin cytoskeleton, focal adhesion dynamics, and tumor cell motility, and identify PHIP as a key driver of glioblastoma migration and invasion.


Assuntos
Neoplasias Encefálicas/patologia , Adesões Focais/patologia , Glioblastoma/patologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Neovascularização Patológica/patologia , Citoesqueleto de Actina/metabolismo , Animais , Encéfalo/patologia , Neoplasias Encefálicas/irrigação sanguínea , Neoplasias Encefálicas/genética , Adesão Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Estudos de Coortes , Progressão da Doença , Feminino , Dosagem de Genes , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Glioblastoma/irrigação sanguínea , Glioblastoma/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Microscopia Intravital , Camundongos , Microscopia Confocal , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Neovascularização Patológica/genética , Imagem com Lapso de Tempo , Vinculina/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Sheng Li Xue Bao ; 72(2): 220-226, 2020 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-32328615

RESUMO

Synaptic cell adhesion molecules (CAMs) are a type of membrane surface glycoproteins that mediate the structural and functional interactions between pre- and post-synaptic sites. Synaptic CAMs dynamically regulate synaptic activity and plasticity, and their expression and function are modulated by environmental factors. Synaptic CAMs are also important effector molecules of stress response, and mediate the adverse impact of stress on cognition and emotion. In this review, we will summarize the recent progress on the role of synaptic CAMs in stress, and aim to provide insight into the molecular mechanisms and drug development of stress-related disorders.


Assuntos
Moléculas de Adesão Celular/fisiologia , Estresse Fisiológico , Estresse Psicológico , Sinapses , Adesão Celular , Humanos , Plasticidade Neuronal
9.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 55(4): 253-258, 2020 Apr 09.
Artigo em Chinês | MEDLINE | ID: mdl-32268625

RESUMO

Objective: To study the effects of titania nanotubes with three different diameters on human gingival fibroblast (HGF). Methods: Three groups of specimens were prepared. Titania nanotubes with diameters of 30, 100, and 200 nm were synthesized on titanium surfaces through electrochemical anodization at 10, 30, and 60 V, respectively. Specimens were assigned into the three groups according to the diameter of the titania nanotubes. Pure smooth titanium without any treatment was set as the control group. HGF were seeded on the surface of the samples. The cell morphology on the specimens was observed with immunofluorescence staining after 2 h, the cell adhesion after 2 d and cell proliferation after 1, 3, and 7 d were detected using methyl thiazolyl tetrazolium assay, and the secretion of type Ⅰ collagen after 7 d was determined using enzyme-linked immunosorbent assay (each group has three samples for each experiment). Results: HGF on the control group exhibited an oval shape without noticeable extensions. HGF on titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm elongated further and were arranged orderly. HGF on titania nanotubes with a diameter of 200 nm were sparsely distributed without noticeable extensions. Titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm could enhance the cell attachment (0.603±0.021 and 0.773±0.045), and secretion of type Ⅰ collagen [(36.5±9.5) and (47.7±4.5) µg/ml, respectively] compared with the control group whose cell attactment was 0.427±0.057, and secretion of type Ⅰ collagen was (22.2±5.9) µg/ml (P<0.05). Furthermore, titania nanotubes with a diameter of 100 nm showed more cell attchment than titania nanotubes with a diameter of 30 nm did (P<0.05). Ttania nanotubes with a diameter of 200 nm clearly impaired the cell adhesion (0.250±0.046) and secretion of type Ⅰ collagen [(10.1±3.7) µg/ml] compared with the control group (P<0.05). At each time point, titania nanotubes with a diameter of 100 nm showed the highest cell proliferation, and the amount of cell proliferation was significantly higher than that on the titania nanotubes with a diameter of 200 nm and the control group at each time point (P<0.05), and was also significantly higher than that on the titania nanotubes with a diameter of 30 nm at day three (P<0.05). At each time point, titania nanotubes with a diameter of 200 nm showed the lowest cell proliferation, which was significantly lower than that on the control group at each time point (P<0.05), except that there was no significant difference in the amount of cell proliferation between titania nanotubes with a diameter of 200 nm and the control group at day one (P>0.05). Conclusions: Titania nanotubes with a diameter of 100 nm can improve the HGF attachment, proliferation, and secretion of type Ⅰ collagen.


Assuntos
Adesão Celular , Proliferação de Células , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Nanotubos , Titânio/farmacologia , Colágeno Tipo I/metabolismo , Humanos , Propriedades de Superfície
10.
PLoS One ; 15(4): e0231752, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32330152

RESUMO

Astrocytes (AC) are the most abundant cells in the central nervous system. In the retina, astrocytes play important roles in the development and integrity of the retinal neurovasculature. Astrocytes dysfunction contributes to pathogenesis of a variety of neurovascular diseases including diabetic retinopathy. Recent studies have demonstrated the expression of Cyp1b1 in the neurovascular cells of the central nervous system including AC. We recently showed retinal AC constitutively express Cyp1b1, and global Cyp1b1-deficiency (Cyp1b1-/-) attenuates retinal ischemia-mediated neovascularization in vivo and the pro-angiogenic activity of retinal vascular cells in vitro. We also demonstrated that Cyp1b1 expression is a key regulator of retinal AC function. However, the underlying mechanisms involved need further investigation. Here we determined changes in the transcriptome profiles of Cyp1b1+/+ and Cyp1b1-/- retinal AC by RNA sequencing. We identified 585 differentially expressed genes, whose pathway enrichment analysis revealed the most significant pathways impacted in Cyp1b1-/- AC. These genes included those of axon guidance, extracellular matrix proteins and their receptors, cancer, cell adhesion molecules, TGF-ß signaling, and the focal adhesion modulation. The expression of a selected set of differentially expressed genes was confirmed by RT-qPCR analysis. To our knowledge, this is the first report of RNAseq investigation of the retinal AC transcriptome and the molecular pathways impacted by Cyp1b1 expression. These results demonstrated an important role for Cyp1b1 expression in the regulation of various retinal AC functions, which are important in neurovascular development and integrity.


Assuntos
Astrócitos/fisiologia , Adesão Celular/genética , Citocromo P-450 CYP1B1/metabolismo , Regulação da Expressão Gênica/fisiologia , Retina/fisiologia , Animais , Movimento Celular/genética , Células Cultivadas , Citocromo P-450 CYP1B1/genética , Feminino , Masculino , Camundongos , Camundongos Knockout , Cultura Primária de Células , RNA-Seq , Retina/citologia
11.
Nat Commun ; 11(1): 1606, 2020 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-32231207

RESUMO

Tetraspanins play critical roles in various physiological processes, ranging from cell adhesion to virus infection. The members of the tetraspanin family have four membrane-spanning domains and short and large extracellular loops, and associate with a broad range of other functional proteins to exert cellular functions. Here we report the crystal structure of CD9 and the cryo-electron microscopic structure of CD9 in complex with its single membrane-spanning partner protein, EWI-2. The reversed cone-like molecular shape of CD9 generates membrane curvature in the crystalline lipid layers, which explains the CD9 localization in regions with high membrane curvature and its implications in membrane remodeling. The molecular interaction between CD9 and EWI-2 is mainly mediated through the small residues in the transmembrane region and protein/lipid interactions, whereas the fertilization assay revealed the critical involvement of the LEL region in the sperm-egg fusion, indicating the different dependency of each binding domain for other partner proteins.


Assuntos
Tetraspanina 29/química , Tetraspanina 29/fisiologia , Animais , Antígenos CD/química , Adesão Celular/fisiologia , Microscopia Crioeletrônica , Cristalografia por Raios X , Feminino , Fertilização/fisiologia , Humanos , Masculino , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Knockout , Modelos Moleculares , Conformação Proteica , Tetraspanina 29/genética
12.
J Biol Regul Homeost Agents ; 34(1 Suppl. 2): 31-35, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32270666

RESUMO

Titanium (Ti) is that the most generally used material for dental, orthopedic and maxillofacial purposes thanks to its excellent biocompatibility and mechanical properties. Several data suggest that prosthesis anchorage to bone and soft tissue are often modulated by surface characteristics. Fibroblasts are the soft tissues cells concerned in producing extracellular matrix and collagen and their tight connection to implant neck is of paramount importance in preventing peri-implant infection. The aim of this work is to grow Human Fibroblast (HFb) for seven days in wells containing (or not) dental implants. The expression levels of some adhesion and traction-resistance related genes (COL11A1, COL2A1, COL9A1, DSP, ELN, HAS1, and TFRC) were analyzed using Polymerase Chain Reaction. Our results demonstrated that several genes encoding for extracellular matrix proteins are activated so giving more insight to the comprehension of the mechanism of cell to surface adhesion.


Assuntos
Adesão Celular , Implantes Dentários , Fibroblastos/citologia , Titânio , Células Cultivadas , Colágeno , Regulação da Expressão Gênica , Humanos , Propriedades de Superfície
13.
Adv Exp Med Biol ; 1221: 309-329, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32274715

RESUMO

Tumor progression associated with hematogenous metastatic spread is a multistep process based on a cross-talk between tumor and stromal cells in a tumor microenvironment. In the blood circulation, tumor cells interact with blood cells through receptors such as selectin and integrins that promote tumor cells survival. At the metastatic sites, heparanase secreted by tumor or stromal cells is an important modifier of the tumor microenvironment while promoting tumor invasiveness and angiogenesis. Heparin, particularly low molecular weight heparin, is used for treatment of cancer patients with evidence of hypercoagulability. However, in preclinical studies heparins was shown to contain other biological activities that affect cancer progression including inhibition of heparanase, selectins and integrins. While ongoing clinical trials are assessing inhibition of heparanase on cancer progression, the remaining biological activities of heparins inhibiting cells adhesion, through selectins and integrins remains largely unexplored. This chapter addresses the potential role of heparins in oncology with respect to their anti-heparanase and anti-adhesive activities and aims to discuss aspects relevant for broader therapeutic application of heparins.


Assuntos
Moléculas de Adesão Celular/antagonistas & inibidores , Glucuronidase/antagonistas & inibidores , Glucuronidase/metabolismo , Heparina/farmacologia , Metástase Neoplásica , Adesão Celular/efeitos dos fármacos , Moléculas de Adesão Celular/metabolismo , Heparina/uso terapêutico , Heparina de Baixo Peso Molecular/farmacologia , Heparina de Baixo Peso Molecular/uso terapêutico , Humanos , Metástase Neoplásica/tratamento farmacológico
14.
BMC Bioinformatics ; 21(1): 95, 2020 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-32126976

RESUMO

BACKGROUND: Many cancers arise from mutations in cells within epithelial tissues. Mutations manifesting at the subcellular level influence the structure and function of the tissue resulting in cancer. Previous work has proposed how cell level properties can lead to mutant cell invasion, but has not incorporated detailed subcellular modelling RESULTS: We present a framework that allows the straightforward integration and simulation of SBML representations of subcellular dynamics within multiscale models of epithelial tissues. This allows us to investigate the effect of mutations in subcellular pathways on the migration of cells within the colorectal crypt. Using multiple models we find that mutations in APC, a key component in the Wnt signalling pathway, can bias neutral drift and can also cause downward invasion of mutant cells in the crypt. CONCLUSIONS: Our framework allows us to investigate how subcellular mutations, i.e. knockouts and knockdowns, affect cell-level properties and the resultant migration of cells within epithelial tissues. In the context of the colorectal crypt, we see that mutations in APC can lead directly to mutant cell invasion.


Assuntos
Neoplasias Colorretais/metabolismo , Modelos Biológicos , Adesão Celular , Movimento Celular , Proliferação de Células , Neoplasias Colorretais/patologia , Bases de Dados Factuais , Humanos , Mutação , Via de Sinalização Wnt
15.
Life Sci ; 249: 117518, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32147432

RESUMO

AIMS: The objectives of the present study were to investigate the mechanisms of Ninj-1 regulation in TNFα-activated human endothelial cells (HEC), and to test if Amlodipine (AML) ameliorates the inflammatory stress by decreasing Ninj-1 expression. MAIN METHODS: TNFα-activated HEC with/without AML (0.1 µM and 1 µM) were used. TNFα-receptor 1 (TNFR1) was silenced and inhibitors for oxidative stress (N-acetyl cysteine), endoplasmic reticulum stress (salubrinal, 4-phenyl butyric acid), or NF-kB (Bay 11-7085) and p38 MAPK (SB203580) were used. Levels of Ninj-1, TNFR1, monocyte adhesion, endoplasmic reticulum stress (ERS) sensors, NADPH oxidase- and mitochondria-derived oxidative species were evaluated. KEY FINDINGS: The novel findings that we report here are: (i) silencing the endothelial TNFR1 leads to decreased Ninj-1 expression and diminished monocyte adhesion; (ii) increased oxidative stress, ERS and NF-kB activation enhance Ninj-1 expression and monocyte adhesion; (iii) up-regulation of endothelial Ninj-1 expression stimulates monocytes adhesion to TNFα - activated HEC; (iv) AML diminishes monocyte adhesion by reducing Ninj-1 expression through mechanisms involving the decrease of NADPH oxidase and mitochondria-dependent oxidative stress, ERS and NF-kB. In addition, AML alleviates apoptosis by reducing the pro-apoptotic CHOP expression and re-establishing the mitochondrial transmembrane potential. SIGNIFICANCE: The results of the present study suggest that Ninj-1 and the proteins involved in its regulation can be considered therapeutic targets for the alleviation of inflammation- dependent disorders. In addition, we demonstrate that some of the benefic effects of AML can be achieved through regulation of Ninj-1.


Assuntos
Anlodipino/farmacologia , Moléculas de Adesão Celular Neuronais/fisiologia , Adesão Celular/fisiologia , Monócitos/citologia , Fatores de Crescimento Neural/fisiologia , Receptores Tipo I de Fatores de Necrose Tumoral/fisiologia , Fator de Necrose Tumoral alfa/fisiologia , Regulação para Cima , Vasodilatadores/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inativação Gênica , Células Endoteliais da Veia Umbilical Humana , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NADPH Oxidases/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética
16.
J Surg Oncol ; 121(7): 1084-1089, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32153051

RESUMO

BACKGROUND AND OBJECTIVES: The aim of this study was to evaluate whether the amount of signet ring cells (SRCs) affects clinicopathological characteristics and prognosis of poorly cohesive (PC) gastric tumours. STUDY DESIGN: One hundred seventy-three patients with PC tumours treated at three European centres from 2004 to 2014 were reclassified in three categories: (a) pure SRC cancers (SRC1) (≥90% SRCs); (b) PC carcinoma with SRC component (SRC2) (>10%, <90% SRCs); (c) PC carcinoma not otherwise specified (SRC3) (≤10% SRCs). RESULTS: The percentage of SRCs was inversely related to the pT stage (Spearman's ρ = -0.174, P < .001) and the number of positive nodes coded as a continuous variable (P = .009). Five year cancer-related survival was significantly higher (58%, 95% confidence interval [CI]: 36%-75%) in SRC1 compared with SRC2 (39%, 95% CI: 28%-50%) and SRC3 (38%, 95% CI: 22%-53%), (P = .048). In multivariable analysis, the impact of PC categories on cancer-related survival was significant when controlling for sex, age, pT, pN, and curativity (hazard ratio [HR] of sSRC2 vs SRC1 = 2.08, 95% CI: 1.01-4.29, P = .046; HR of SRC3 vs SRC1 = 2.38, 95% CI: 1.05-5.41, P = .039). CONCLUSION: The percentage of SRCs was inversely related to tumour aggressiveness, with long-term survival significantly higher in SRC1 compared with SRC2 and SRC3 tumours.


Assuntos
Carcinoma de Células em Anel de Sinete/patologia , Neoplasias Gástricas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células em Anel de Sinete/mortalidade , Adesão Celular/fisiologia , Europa (Continente)/epidemiologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Retrospectivos , Neoplasias Gástricas/mortalidade
17.
Nat Commun ; 11(1): 1143, 2020 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-32123168

RESUMO

By offering the possibility to manipulate cellular functions with spatiotemporal control, optogenetics represents an attractive tool for dissecting immune responses. However, applying these approaches to single cells in vivo remains particularly challenging for immune cells that are typically located in scattering tissues. Here, we introduce an improved calcium actuator with sensitivity allowing for two-photon photoactivation. Furthermore, we identify an actuator/reporter combination that permits the simultaneous manipulation and visualization of calcium signals in individual T cells in vivo. With this strategy, we document the consequences of defined patterns of calcium signals on T cell migration, adhesion, and chemokine release. Manipulation of individual immune cells in vivo should open new avenues for establishing the functional contribution of single immune cells engaged in complex reactions.


Assuntos
Sinalização do Cálcio/fisiologia , Optogenética/métodos , Linfócitos T/metabolismo , Animais , Proteínas de Arabidopsis/genética , Linfócitos T CD8-Positivos/metabolismo , Adesão Celular , Movimento Celular , Quimiocinas/metabolismo , Criptocromos/genética , Células HEK293 , Humanos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fótons , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Célula Única/métodos , Molécula 1 de Interação Estromal/genética , Molécula 1 de Interação Estromal/metabolismo , Linfócitos T/citologia
18.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(2): 165-170, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32220183

RESUMO

Objective: To investigate the regulation of fibromodulin (FMOD) on proliferation, adhesion and migration of non-small cell lung cancer cell line H322, and discuss its action mechanism. Methods: H322 cells were randomly divided into control group, small interfering RNA (siRNA) silencing FMOD ( FMOD siRNA) group and control siRNA (Con siRNA) group. FMOD siRNA and Con siRNA were transfected into H322 cells. The cell viability of each group was detected by CCK-8 method. The adhesion ability of cells was detected by fluorescein diacetate (FDA) fluorescent staining. The cell migration ability was detected by Transwell method. Real time-PCR was used to detect the mRNA expressions of Cyclin D1, intercellular adhesion molecule -1 (ICAM-1), E-cadherin, FMOD, transforming growth factor-ß (TGF-ß), Smad2, Smad3, Smad4 and Smad7 in cells. The protein expressions of Cyclin D1, ICAM-1, E-cadherin, FMOD, TGF-ß1, Smad2, Smad3, Smad4 and Smad7 were detected by Western blot. Results: Compared with the Con siRNA group, the cell viability, cell adhesion and migration ability of the FMOD siRNA group were decreased, and the difference was statistically significant ( P<0.01). There was no significant difference between the control group and the Con siRNA group. Real time-PCR and Western blot results showed that the mRNA and protein expression levels of Cyclin D1, ICAM-1, TGF-ß1, Smad2, Smad3 and Smad4 were decreased in FMOD siRNA group, compared with Con siRNA group, while the mRNA and protein expression levels of E-cadherin and Smad7 are elevated. Conclusion: Silencing of the FMOD gene significantly reduces the proliferation, adhesion and migration of H322 cells, which may be conducted by inhibiting the TGF-ß/Smad signaling pathway.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Fibromodulina/genética , Inativação Gênica , Neoplasias Pulmonares , Proteínas Smad , Fator de Crescimento Transformador beta , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Adesão Celular , Movimento Celular , Proliferação de Células , Fibromodulina/fisiologia , Expressão Gênica , Humanos , Neoplasias Pulmonares/metabolismo , RNA Interferente Pequeno , Transdução de Sinais , Proteínas Smad/fisiologia , Fator de Crescimento Transformador beta/fisiologia
19.
Nat Cell Biol ; 22(4): 498-511, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32203420

RESUMO

Rho GTPases are central regulators of the cytoskeleton and, in humans, are controlled by 145 multidomain guanine nucleotide exchange factors (RhoGEFs) and GTPase-activating proteins (RhoGAPs). How Rho signalling patterns are established in dynamic cell spaces to control cellular morphogenesis is unclear. Through a family-wide characterization of substrate specificities, interactomes and localization, we reveal at the systems level how RhoGEFs and RhoGAPs contextualize and spatiotemporally control Rho signalling. These proteins are widely autoinhibited to allow local regulation, form complexes to jointly coordinate their networks and provide positional information for signalling. RhoGAPs are more promiscuous than RhoGEFs to confine Rho activity gradients. Our resource enabled us to uncover a multi-RhoGEF complex downstream of G-protein-coupled receptors controlling CDC42-RHOA crosstalk. Moreover, we show that integrin adhesions spatially segregate GEFs and GAPs to shape RAC1 activity zones in response to mechanical cues. This mechanism controls the protrusion and contraction dynamics fundamental to cell motility. Our systems analysis of Rho regulators is key to revealing emergent organization principles of Rho signalling.


Assuntos
Citoesqueleto/genética , Proteínas Ativadoras de GTPase/genética , Integrinas/genética , Mecanotransdução Celular/genética , Fatores de Troca de Nucleotídeo Guanina Rho/genética , Proteínas rac1 de Ligação ao GTP/genética , Animais , Células COS , Adesão Celular , Linhagem Celular , Movimento Celular , Chlorocebus aethiops , Biologia Computacional , Citoesqueleto/metabolismo , Citoesqueleto/ultraestrutura , Cães , Fibroblastos/metabolismo , Fibroblastos/ultraestrutura , Proteínas Ativadoras de GTPase/classificação , Proteínas Ativadoras de GTPase/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Células HEK293 , Células HeLa , Humanos , Integrinas/metabolismo , Células Madin Darby de Rim Canino , Camundongos , Pan troglodytes , Domínios Proteicos , Ratos , Fatores de Troca de Nucleotídeo Guanina Rho/classificação , Fatores de Troca de Nucleotídeo Guanina Rho/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo
20.
Environ Sci Pollut Res Int ; 27(15): 17770-17778, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32162219

RESUMO

Pardosa pseudoannulata (Araneae: Lycosidae), as an important predator of crop pests, has served as a strong driver for ecological regulation of pests. Cadmium (Cd) is a toxic heavy metal widely distributed in the soil in China, which not only seriously pollutes the ecological environment, but also poses a great threat to the survival of organisms. Palpal bulbs are the genital organs of male spiders, playing an important role in reproductive physiology. However, the effects of long-term Cd stress on the genital organ of the primary pest predator were poorly understood. Therefore, we investigated the Cd effect on the male palpal organ of P. pseudoannulata at morphological and gene expression levels. The results showed that no obvious difference in the morphology between the Cd-treated and control groups was observed, but cell adhesion was affected at molecular level. Transcriptome sequencing analysis revealed that under long-term Cd stress, the biological processes including cell-cell adhesion via plasma-membrane adhesion molecules, cell-cell adhesion, and homophilic cell adhesion via plasma membrane adhesion molecules were the top three differentially expressed terms (p-adj < 0.001), and 51 unigenes were annotated into cadherin-related proteins, such as protocadherin, cadherin-87A, and cadherin-96Ca, among which, 18 unigenes were significantly upregulated under the Cd stress. Our outcomes indicate that the differentially expressed genes involved in cell adhesion may explain the negative effects of Cd stress on the spider genital organ, and the comprehensive transcriptome dataset will also provide a profound molecular information of the genital organ of P. pseudoannulata.


Assuntos
Cádmio , Aranhas/genética , Animais , Caderinas , Adesão Celular , China , Genitália Masculina , Masculino , Transcriptoma
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