Chronic Resveratrol Treatment Ameliorates Cell Adhesion and Mitigates the Inflammatory Phenotype in Senescent Human Fibroblasts

Chondroadherin (the 36-kD protein) is a leucine-rich, cartilage matrix protein known to mediate adhesion of isolated chondrocytes. In the present study we investigated cell surface proteins involved in the interaction of cells with chondroadherin in cell adhesion and by affinity purification. Adhesion of bovine articular chondrocytes to chondroadherin-coated dishes was dependent on Mg2+ or Mn2+ but not Ca2+. Adhesion was partially inhibited by an antibody recognizing β1 integrin subunit. Chondroadherin-binding proteins from chondrocyte lysates were affinity purified on chondroadherin-Sepharose. The β1 integrin antibody immunoprecipitated two proteins with molecular mass ∼110 and 140 kD (nonreduced) from the EDTA-eluted material. These results indicate that a β1 integrin on chondrocytes interacts with chondroadherin. To identify the α integrin subunit(s) involved in interaction of cells with the protein, we affinity purified chondroadherin-binding membrane proteins from human fibroblasts. Immunoprecipitation of the EDTA-eluted material from the affinity column identified α2β1 as a chondroadherin-binding integrin. These results are in agreement with cell adhesion experiments where antibodies against the integrin subunit α2 partially inhibited adhesion of human fibroblast and human chondrocytes to chondroadherin. Since α2β1 also is a receptor for collagen type II, we tested the ability of different antibodies against the α2 subunit to inhibit adhesion of T47D cells to collagen type II and chondroadherin. The results suggested that adhesion to collagen type II and chondroadherin involves similar or nearby sites on the α2β1 integrin. Although α2β1 is a receptor for both collagen type II and chondroadherin, only adhesion of cells to collagen type II was found to mediate spreading.

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Cytoskeletal integrity is required throughout the mitogen stimulation phase of the cell cycle and mediates the anchorage-dependent expression of cyclin D1.

Molecular Biology of the Cell ◽

10.1091/mbc.7.1.101 ◽

1996 ◽

Vol 7 (1) ◽

pp. 101-111 ◽

Cited By ~ 147

Author(s):

R M Böhmer ◽

E Scharf ◽

R K Assoian

Keyword(s):

Cell Cycle ◽

Cell Adhesion ◽

Cyclin D1 ◽

Cell Cycle Progression ◽

Human Fibroblasts ◽

Cytochalasin D ◽

Phase Cell ◽

Dependent Part ◽

Molecular Events ◽

Normal Human

The proliferation of many nontransformed cells depends on cell adhesion. We report here that disrupting the cytoskeleton in normal human fibroblasts causes the same cell cycle phenotype that is observed after blocking cell adhesion: suspended cells and cytochalasin D-treated monolayers fail to progress through G1 despite normal mitogen-induced expression of c-myc mRNA. Midway between G0 and the beginning of S-phase, cell cycle progression becomes independent of adhesion and the cytoskeleton. At this stage, the cells are also mitogen independent. Molecular analyses showed that Rb hyperphosphorylation and the induction of cyclin D1 occur slightly earlier than the transition to cytoskeleton independence. Moreover, these molecular events are blocked by cytochalasin D. Overall, our data indicate the following: 1) anchorage and cytoskeletal integrity are required throughout the mitogen-dependent part of G1; 2) mitogens and the cytoskeleton jointly regulate the phosphorylation of Rb; and 3) this interdependence is manifest in the regulation of cyclin D1.

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Focal Adhesion Kinase Modulates Cell Adhesion Strengthening via Integrin Activation

Molecular Biology of the Cell ◽

10.1091/mbc.e08-01-0076 ◽

2009 ◽

Vol 20 (9) ◽

pp. 2508-2519 ◽

Cited By ~ 139

Author(s):

Kristin E. Michael ◽

David W. Dumbauld ◽

Kellie L. Burns ◽

Steven K. Hanks ◽

Andrés J. García

Keyword(s):

Cell Migration ◽

Cell Adhesion ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Human Fibroblasts ◽

Inducible Promoter ◽

Time Dependent ◽

Integrin Activation ◽

Nonreceptor Tyrosine Kinase ◽

Adhesive Forces

Focal adhesion kinase (FAK) is an essential nonreceptor tyrosine kinase regulating cell migration, adhesive signaling, and mechanosensing. Using FAK-null cells expressing FAK under an inducible promoter, we demonstrate that FAK regulates the time-dependent generation of adhesive forces. During the early stages of adhesion, FAK expression in FAK-null cells enhances integrin activation to promote integrin binding and, hence, the adhesion strengthening rate. Importantly, FAK expression regulated integrin activation, and talin was required for the FAK-dependent effects. A role for FAK in integrin activation was confirmed in human fibroblasts with knocked-down FAK expression. The FAK autophosphorylation Y397 site was required for the enhancements in adhesion strengthening and integrin-binding responses. This work demonstrates a novel role for FAK in integrin activation and the time-dependent generation of cell–ECM forces.

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Mast Cells Induce T-Cell Adhesion to Human Fibroblasts by Regulating Intercellular Adhesion Molecule-1 and Vascular Cell Adhesion with Expression

Journal of Investigative Dermatology ◽

10.1111/1523-1747.ep12326075 ◽

1995 ◽

Vol 105 (6) ◽

pp. 789-796 ◽

Cited By ~ 29

Author(s):

Hong Meng ◽

Mary J. Marchese ◽

Jonathan A. Garlick ◽

Ante Jelaska ◽

Joseph H. Korn ◽

...

Keyword(s):

Cell Adhesion ◽

T Cell ◽

Mast Cells ◽

Adhesion Molecule ◽

Human Fibroblasts ◽

Intercellular Adhesion ◽

Intercellular Adhesion Molecule ◽

Vascular Cell Adhesion ◽

Intercellular Adhesion Molecule 1 ◽

Vascular Cell

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Modulation of the Senescence-Associated Inflammatory Phenotype in Human Fibroblasts by Olive Phenols

International Journal of Molecular Sciences ◽

10.3390/ijms18112275 ◽

2017 ◽

Vol 18 (11) ◽

pp. 2275 ◽

Cited By ~ 13

Author(s):

Beatrice Menicacci ◽

Caterina Cipriani ◽

Francesca Margheri ◽

Alessandra Mocali ◽

Lisa Giovannelli

Keyword(s):

Human Fibroblasts ◽

Inflammatory Phenotype

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Chimpanzee fibroblasts exhibit greater adherence and migratory phenotypes than human fibroblasts

10.1101/838755 ◽

2019 ◽

Author(s):

Trisha M. Zintel ◽

Delaney Ducey ◽

Courtney C. Babbitt

Keyword(s):

Gene Expression ◽

Wound Healing ◽

Cell Adhesion ◽

Cell Lines ◽

Focal Adhesion ◽

Cell Biology ◽

Human Fibroblasts ◽

Cellular Adhesion ◽

Specific Cell ◽

And Migration

ABSTRACTBackground and objectivesPrevious work has identified that gene expression differences in cell adhesion pathways exist between humans and chimpanzees. Here, we used a comparative cell biology approach to assay interspecies differences in cell adhesion phenotypes in order to better understand the basic biological differences between species’ epithelial cells that may underly the organism-level differences we see in wound healing and cancer.MethodologyWe used skin fibroblast cell lines from humans and chimpanzees to assay cell adhesion and migration. We then utilized published RNA-Seq data from the same cell lines exposed to a cancer / wound-healing mimic to determine what gene expression changes may be corresponding to altered cellular adhesion dynamics between species.ResultsThe functional adhesion and migration assays revealed that chimpanzee fibroblasts adhered sooner and remained adherent for significantly longer and move into a “wound” at faster rate than human fibroblasts. The gene expression data suggest that the enhanced adhesive properties of chimpanzee fibroblasts may be due to chimpanzee fibroblasts exhibiting significantly higher expression of cell and focal adhesion molecule genes than human cells, both during a wound healing assay and at rest.Conclusions and implicationsChimpanzee fibroblasts exhibit stronger adhesion and greater cell migration than human fibroblasts. This may be due to divergent gene expression of focal adhesion and cell adhesion molecules, such as integrins, laminins, and cadherins, as well as ECM proteins like collagens. This is one of few studies demonstrating that these divergences in gene expression between closely related species can manifest in fundamental differences in cell biology. Our results provide better insight into species-specific cell biology phenotypes and how they may influence more complex traits, such as cancer metastasis and wound healing.

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Stem-like Cancer Cells in a Dynamic 3D Culture System: A Model to Study Metastatic Cell Adhesion and Anti-cancer Drugs

Cells ◽

10.3390/cells8111434 ◽

2019 ◽

Vol 8 (11) ◽

pp. 1434 ◽

Cited By ~ 4

Author(s):

Paolillo ◽

Colombo ◽

Serra ◽

Belvisi ◽

Papetti ◽

...

Keyword(s):

Cell Adhesion ◽

Cancer Cells ◽

Culture System ◽

Human Fibroblasts ◽

3D Culture ◽

Target Tissue ◽

Cancer Drugs ◽

Mesenchymal Transition ◽

Anti Cancer

Metastatic spread is mainly sustained by cancer stem cells (CSC), a subpopulation of cancer cells that displays stemness features. CSC are thought to be derived from cancer cells that undergo epithelial to mesenchymal transition (EMT), thus acquiring resistance to anoikis and anti-cancer drugs. After detachment from the primary tumor mass, CSC reach the blood and lymphatic flow, and disseminate to the target tissue. This process is by nature dynamic and in vitro models are quite far from the in vivo situation. In this study, we have tried to reproduce the adhesion process of CSC to a target tissue by using a 3D dynamic cell culture system. We isolated two populations of 3D tumor spheroids displaying CSC-like features from breast carcinoma (MCF-7) and lung carcinoma (A549) cell lines. Human fibroblasts were layered on a polystyrene scaffold placed in a dynamically perfused millifluidic system and then the adhesion of tumor cell derived from spheroids to fibroblasts was investigated under continuous perfusion. After 24 h of perfusion, we found that spheroid cells tightly adhered to fibroblasts layered on the scaffold, as assessed by a scanning electron microscope (SEM). To further investigate mechanisms involved in spheroid cell adhesion to fibroblasts, we tested the effect of three RGD integrin antagonists with different molecular structures on cell adhesion; when injected into the circuit, only cilengitide was able to inhibit cell adhesion to fibroblasts. Although our model needs further refinements and improvements, we do believe this study could represent a promising approach in improving current models to study metastatic infiltration in vitro and a new tool to screen new potential anti-metastatic molecules.

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Dual functions of transglutaminase in novel cell adhesion

Journal of Cell Science ◽

10.1242/jcs.109.11.2727 ◽

1996 ◽

Vol 109 (11) ◽

pp. 2727-2735 ◽

Cited By ~ 1

Author(s):

S. Ueki ◽

J. Takagi ◽

Y. Saito

Keyword(s):

Cell Adhesion ◽

Enzyme Preparation ◽

Human Fibroblasts ◽

Coagulation Factor ◽

Factor Xiiia ◽

Ammonium Ion ◽

Dependent Manner ◽

Tgase Activity ◽

Adhesion Activity ◽

First Time

Transglutaminases (TGases) are enzymes which catalyze cross-link formation between glutamine residues and lysine residues in substrate proteins. In the present study, we report for the first time that a representative enzyme, blood coagulation factor XIIIa (FXIIIa), is capable of mediating adhesion of various cells. When coated on plastic surfaces FXIIIa promoted adhesion and spreading of various cells of both normal and tumor origin, in a concentration-dependent manner. The adhesion was not inhibited by antibodies against possible contaminants in the enzyme preparation such as fibronectin and vitronectin, but was completely inhibited by a polyclonal antibody against the enzyme. Therefore, if there were any contaminating cell adhesive substrates in the enzyme preparation, they cannot account for the observed cell adhesion to the enzyme; FXIIIa itself mediates the cell adhesion. Furthermore, phosphorylation of tyrosine residues in 120 kDa and 70 kDa proteins was clearly shown in human fibroblasts adhering to the enzyme. Formation of actin stress fibers was also unambiguously observed in the adhering cells. These biochemical reactions, which are also observed when cells adhere to a typical cell adhesion protein, fibronectin, are believed to be of importance in the process of cell adhesion. This adhesion activity of FXIIIa was dependent on its TGase activity, because both a modification of the active center cysteine with iodoacetamide and the addition of ammonium ion abolished the cell adhesion activity along with the enzyme activity. The cell adhesion to fibronectin, however, was not affected by these treatments. The effects of various anti-integrin antibodies suggested that both alpha v beta 3 and beta 1 family integrins participated in the cell adhesion to FXIIIa. Taken together, these data demonstrate for the first time that there is a unique TGase activity-mediated cell adhesion. This novel function of the enzyme may be of physiological importance.

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