DICAM, a novel dual immunoglobulin domain containing cell adhesion molecule interacts with alphavbeta3 integrin

Immunoglobulin (Ig) superfamily members are abundant with diverse functions including cell adhesion in various tissues. Here, we identified and characterized a novel adhesion molecule that belongs to the CTX protein family and named as DICAM (Dual Ig domain containing cell adhesion molecule). DICAM is a type I transmembrane protein with two V-type Ig domains in the extracellular region and a short cytoplasmic tail of 442 amino acids. DICAM is found to be expressed ubiquitously in various organs and cell lines. Subcellular localization of DICAM was observed in the cell-cell contact region and nucleus of cultured epithelial cells. Cell-cell contact region was colocalized with tight junction protein, ZO-1. The DICAM increased MDCK cell adhesion to 60% levels of fibronectin. DICAM mediated cell adhesion was specific for the alphavbeta3 integrin; other integrins, alpha2, alpha5, beta1, alpha2beta1, alpha5beta1, were not involved in cell adhesion. In identifying the interacting domain of DICAM with alphavbeta3, the Ig domain 2 showed higher cell adhesion activity than that of Ig domain 1. Although RGD motif in Ig domain 2 was engaged in cell adhesion, it was not participated in DICAM-alphavbeta3 mediated cell adhesion. Furthermore, differentially expressing DICAM stable cells showed well correlated cell to cell adhesion capability with integrin beta3-overexpressing cells. Collectively, these results indicate that DICAM, a novel dual Ig domain containing adhesion molecule, mediates cell adhesion via alphavbeta3 integrin.     

The roles of cell adhesion molecules in tumor suppression and cell migration

In addition to mediating cell adhesion, many cell adhesion molecules act as tumor suppressors. These proteins are capable of restricting cell growth mainly through contact inhibition. Alterations of these cell adhesion molecules are a common event in cancer. The resulting loss of cell-cell and/or cell-extracellular matrix adhesion promotes cell growth as well as tumor dissemination. Therefore, it is conventionally accepted that cell adhesion molecules that function as tumor suppressors are also involved in limiting tumor cell migration. Paradoxically, in 2005, we identified an immunoglobulin superfamily cell adhesion molecule hepaCAM that is able to suppress cancer cell growth and yet induce migration. Almost concurrently, CEACAM1 was verified to co-function as a tumor suppressor and invasion promoter. To date, the reason and mechanism responsible for this exceptional phenomenon remain unclear. Nevertheless, the emergence of these intriguing cell adhesion molecules with conflicting roles may open a new chapter to the biological significance of cell adhesion molecules.     

TRPM7 regulates cell adhesion by controlling the calcium-dependent protease calpain

m-Calpain is a protease implicated in the control of cell adhesion through focal adhesion disassembly. The mechanism by which the enzyme is spatially and temporally controlled is not well understood, particularly because the dependence of calpain on calcium exceeds the submicromolar concentrations normally observed in cells. Here we show that the channel kinase TRPM7 localizes to peripheral adhesion complexes with m-calpain, where it regulates cell adhesion by controlling the activity of the protease. Our research revealed that overexpression of TRPM7 in cells caused cell rounding with a concomitant loss of cell adhesion that is dependent upon the channel of the protein but not its kinase activities. Knockdown of m-calpain blocked TRPM7-induced cell rounding and cell detachment. Silencing of TRPM7 by RNA interference, however, strengthened cell adhesion and increased the number of peripheral adhesion complexes in the cells. Together, our results suggest that the ion channel TRPM7 regulates cell adhesion through m-calpain by mediating the local influx of calcium into peripheral adhesion complexes.     

The adhesion of BHK and PyBHK cells to the substratum

The adhesion of BHK 21/13 cells and their polyoma-transformed derivatives was studied by detaching them from plastic dishes with EGTA. The PyBHK cells were less adhesive, and the possibility that cyclic nucleotides might play a role in cell adhesion was examined. Both cAMP and cGMP increase cell adhesion through a mechanism involving microtubules, but glucocorticoids act to increase cell adhesion through an independent mechanism.We also examined the role of membrane fluidity in cell adhesion, and the results are discussed in terms of a general model for cell adhesion and locomotion.     

Effect of central myelin on the proliferation and differentiation into O4+ oligodendrocytes of GFP-NSCs

Cell adhesion is an important process during morphogenesis, differentiation, and homeostasis in cell biology. The role of vascular endothelial growth factor (VEGF) in cell adhesion of keratinocytes is unclear. In our study, a human keratinocyte cell line, HaCaT cells, which mimics various properties of normal epidermal keratinocytes, was included to elucidate the effect of VEGF on cell-cell adhesion and cell-plate adhesion. Expression of adhesion molecules account for cell adhesion and signal transduction pathways involved in the effect of VEGF on adhesion of HaCaT cells were further investigated. Significant increase of cell-cell adhesion but decrease of the cell-plate adhesion of HaCaT cells induced by VEGF(165) was detected. VEGF increases expression of E-cadherin, but inhibits expression of integrin α6β4 subunit. VEGF(165) at 100?ng/ml activates extracellular signal-regulated kinase. These changes of cell adhesion induced by VEGF were blocked by ERK and VEGFR-2 inhibitor. Our findings suggest that VEGF may modulate cell adhesion of HaCaT cells partly through activation of VEGFR-2/ERK1/2 signaling pathways.     

Activity-independent cell adhesion to tissue-type transglutaminase is mediated by alpha4beta1 integrin

Transglutaminases (TGases) are enzymes which catalyze cross-link formation between glutamine residues and lysine residues in substrate proteins. We have previously reported that one of the TGases, blood coagulation factor XIIIa (FXIIIa), is capable of mediating adhesion of various cells. In this paper, we report for the first time that tissue-type transglutaminase (TGc) also has cell adhesion activity. TGc-coated plastic surface promoted adhesion and spreading of cells in a TGc concentration-dependent manner. However, there are some obvious differences between cell adhesion mediated by TGc and FXIIIa. As was reported previously, the adhesion to FXIIIa is dependent on its TGase activity. In contrast, the TGc-mediated cell adhesion is independent of its TGase activity: 1) The modification of the active center cysteine with iodoacetamide blocked the enzyme activity without any effect on cell adhesion; 2) the addition of Mg2+ did not induce the enzyme activity, but it was as effective as Ca2+ for cell adhesion; 3) the addition of NH4+ inhibited the enzyme activity but did not affect the cell adhesion significantly. The integrins involved in these cell adhesions are quite different. In the case of FXIIIa, alpha vbeta3 and alpha5beta1 integrins are involved and consequently the RGD peptide substantially inhibited the adhesion. On the other hand, the cell adhesion to TGc is mediated by alpha4beta1 integrin but not alpha5beta1; a CS-1 peptide, which represents the binding site of fibronectin to alpha4beta1 integrin, completely inhibited the cell adhesion to TGc. It is possible that TGc and FXIIIa may mediate cell adhesion under different physiological and pathological situations.     

The relation of temperature and lipid composition to cell adhesion

We have examined as a function of temperature the effect of changes in the composition of the fatty acid chains of membrane phospholipids on the rate of cell to cell adhesion in the neuronal cell line B103. The rate of cell to cell adhesion in this cell line is highly temperature dependent but is not influenced by changes in the fatty acid composition of the plasma membrane generated by growing the cells either in the presence of oleic acid or elaidic acid. In contrast the temperature dependence of the rate of cell to cell adhesion, measured in a monolayer adhesion assay, is highly dependent on the shear force used during the assay. A two-step model of cell to cell adhesion involving multiple adhesion ligands is presented which can be used to explain these observations.     

Phosphorylation of Trask by Src kinases inhibits integrin clustering and functions in exclusion with focal adhesion signaling

Trask is a recently described transmembrane substrate of Src kinases whose expression and phosphorylation has been correlated with the biology of some cancers. Little is known about the molecular functions of Trask, although its phosphorylation has been associated with cell adhesion. We have studied the effects of Trask phosphorylation on cell adhesion, integrin activation, clustering, and focal adhesion signaling. The small hairpin RNA (shRNA) knockdown of Trask results in increased cell adhesiveness and a failure to properly inactivate focal adhesion signaling, even in the unanchored state. On the contrary, the experimentally induced phosphorylation of Trask results in the inhibition of cell adhesion and inhibition of focal adhesion signaling. This is mediated through the inhibition of integrin clustering without affecting integrin affinity state or ligand binding activity. Furthermore, Trask signaling and focal adhesion signaling inactivate each other and signal in exclusion with each other, constituting a switch that underlies cell anchorage state. These data provide considerable insight into how Trask functions to regulate cell adhesion and reveal a novel pathway through which Src kinases can oppose integrin-mediated cell adhesion.     

The relation of temperature and lipid composition to cell adhesion.

We have examined as a function of temperature the effect of changes in the composition of the fatty acid chains of membrane phospholipids on the rate of cell to cell adhesion in the neuronal cell line B103. The rate of cell to cell adhesion in this cell line is highly temperature dependent but is not influenced by changes in the fatty acid composition of the plasma membrane generated by growing the cells either in the presence of oleic acid or elaidic acid. In contrast the temperature dependence of the rate of cell to cell adhesion, measured in a monolayer adhesion assay, is highly dependent on the shear force used during the assay. A two-step model of cell to cell adhesion involving multiple adhesion ligands is presented which can be used to explain these observations.     

Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Tissue‐embedded cells are often exposed to a complex mixture of extracellular matrix (ECM) molecules, to which they bind with different cell adhesion receptors and affinities. Differential cell adhesion to ECM components is believed to regulate many aspects of tissue function, such as the sorting of specific cell types into different tissue compartments or ECM niches. In turn, aberrant switches in cell adhesion preferences may contribute to cell misplacement, tissue invasion, and metastasis. Methods to determine differential adhesion profiles of single cells are therefore desirable, but established bulk assays usually only test cell population adhesion to a single type of ECM molecule. We have recently demonstrated that atomic force microscopy‐based single‐cell force spectroscopy (SCFS), performed on bifunctional, microstructured adhesion substrates, provides a useful tool for accurately quantitating differential matrix adhesion of single Chinese hamster ovary cells to laminin and collagen I. Here, we have extended this approach to include additional ECM substrates, such as bifunctional collagen I/collagen IV surfaces, as well as adhesion‐passivated control surfaces. We investigate differential single cell adhesion to these substrates and analyze in detail suitable experimental conditions for comparative SCFS, including optimal cell‐substrate contact times and the impact of force cycle repetitions on single cell adhesion force statistics. Insight gained through these experiments may help in adapting this technique to other ECM molecules and cell systems, making directly comparative SCFS a versatile tool for comparing receptor‐mediated cell adhesion to different matrix molecules in a wide range of biological contexts. Copyright © 2013 John Wiley & Sons, Ltd.     

CTL adhesion and antigen recognition are discrete steps in the human CTL-target cell interaction

Th initial step in cytolytic T lymphocyte (CTL)-mediated cytolysis involves target cell adhesion and antigen recognition. To investigate these initial events in the CTL-target interaction, we used HLA-A2- and HLA-B7-specific human CTL clones and HLA-typed B lymphoblastoid target cells. By using two different adhesion assays, we demonstrated antigen nonspecific CTL-target cell adhesion. To more precisely define the contribution of the antigen-specific receptor to CTL-target cell adhesion, we used the HLA-A2, HLA-B7, and mock transfected RD target cells. Consistent with the results when using B lymphoblastoid target cells, the CTL clones demonstrated equivalent adhesions to the RD target cells whether or not they expressed HLA-A2 or HLA-B7. These results suggested that CTL-target cell adhesion occurred independent of the T cell receptor. By using the calcium-sensitive dye Indo-1 and flow cytometry, we assessed CTL-target cell adhesion and CTL activation. Simultaneous measurement of adhesion and intracellular free calcium demonstrated that CTL-target cell adhesion alone did not activate CTL clones. Both CTL-target cell adhesion and the presence of the appropriate HLA target molecule were necessary for the efficient activation of human CTL. MAb inhibition studies indicated that antigen nonspecific adhesion is largely regulated by the LFA-1, CD2 (LFA-2/T11), and LFA-3 cell surface molecules. These antigen nonspecific cell-cell interaction molecules appear to play an important role in facilitating antigen recognition and subsequent target cell lysis.     

白细胞与内皮细胞的粘附

白细胞与内皮细胞相互作用由粘附分子介导.整合素、免疫球蛋白及选择素家族的粘附分子在这两种细胞的粘附中起关键作用.粘附的起始阶段由选择素介导,随后由CD11/CD18复合物与ICAM-1形成更为紧密的结合.多种细胞因子及炎症反应可诱导粘附.抗粘附分子单抗、药物等可抑制粘附.     

Flow-conditioned HUVECs support clustered leukocyte adhesion by coexpressing ICAM-1 and E-selectin

Endothelial sequestration of circulating monocytes is a key event in early atherosclerosis. Hemodynamics is proposed to regulate monocyte-endothelial cell interactions by direct cell activation and establishment of flow environments that are conducive or prohibitive to cell-cell interaction. We investigated fluid shear regulation of monocyte-endothelial cell adhesion in vitro using a disturbed laminar shear system that models in vivo hemodynamics characteristic of lesion-prone vascular regions. Human endothelial cell monolayers were flow conditioned for 6 h before evaluation of monocyte adhesion under static and dynamic flow conditions. Results revealed a distinctive clustered cell pattern of monocyte adhesion that strongly resembles in vivo leukocyte adhesion in early- and late-stage atherosclerosis. Clustered monocyte cell adhesion correlated with endothelial cells coexpressing intercellular adhesion molecule-1 (ICAM-1) and E-selectin as result of a flow-induced, selective upregulation of E-selectin expression in a subset of ICAM-1-expressing cells. Clustered monocyte cell adhesion assayed under static conditions exhibited a spatial variation in size and frequency of occurrence, which demonstrates differential regulation of endothelial cell adhesiveness by the local flow environment. Dynamic adhesion studies conducted with circulating monocytes resulted in clustered cell adhesion only within the disturbed flow region, where the monocyte rate of motion is sufficiently low for cell-cell interaction. These studies provide evidence and reveal mechanisms of local hemodynamic regulation of endothelial adhesiveness and endothelial monocyte interaction that lead to localized monocyte adhesion and potentially contribute to the focal origin of arterial diseases such as atherosclerosis.     

紫锥菊多糖抑制致病性大肠埃希菌细胞黏附的试验研究

目的研究紫锥菊多糖能否影响致病性大肠埃希菌对细胞的黏附。方法使用PK-15细胞进行黏附试验及黏附抑制试验。结果发现紫锥菊多糖浓度为1.6 mg/ml时,对细菌黏附细胞的抑制作用最好,黏附率由50个细菌/细胞降低到6.8个细菌/细胞。结论紫锥菊多糖对致病性大肠埃希菌的细胞黏附具有抑制作用,提示该多糖具有调节肠道微生态的潜在应用价值。     

板蓝根多糖抑制致病性大肠埃希菌细胞黏附的试验研究

研究板蓝根多糖能否影响致病性大肠埃希菌对细胞的黏附。使用PK-15细胞进行了黏附试验及黏附抑制试验。在所选的4个浓度中,板蓝根多糖浓度为1.6mg/mL时,对细菌黏附细胞的抑制作用最好,黏附力由每个细胞黏附44.8个细菌降低到6.3个细菌。板蓝根多糖对致病性大肠埃希菌的细胞黏附具有抑制作用,提示该多糖具有调节肠道微生态的潜在应用价值。     

Expression level of neural cell adhesion molecule (NCAM) inversely correlates with the ability of neuroblastoma cells to adhere to endothelium in vitro

The precise function of cell adhesion molecules in the hematogenous phase of neuroblastoma metastasis is poorly understood. The aim of this study was to investigate whether neural cell adhesion molecule (NCAM) modulates neuroblastoma cell (NB) adhesion and transendothelial penetration in a coculture model. Our data, assessed on 11 NB cell lines, demonstrate an inverse correlation between NCAM expression and NB cell adhesion. Transfection of the NB cell line UKF-NB-4 with a cDNA encoding the human NCAM-140 kD isoform enhanced NCAM expression and the amount of tumor cell aggregates, reduced the amount of single tumor cells, and diminished initial NB cell adhesion to an endothelial cell monolayer. Treatment of UKF-NB-4 with NCAM antisense oligonucleotides reduced NCAM surface level, increased the number of single tumor cells, and induced up-regulation of NB cell adhesion to endothelium. Modulation of NCAM expression had no effect on transendothelial penetration. Fluorescence analysis revealed a down-regulation of NCAM in single tumor cells, prior to NB adhesion. The data support the view that low levels of NCAM are necessary for NB cells to leave a tumor cell aggregate and adhere to endothelial cells.     

蛋白酶活化受体2促进结肠癌细胞与基质粘附

目的:蛋白酶活化受体2激活对结肠癌细胞HT29粘附能力的影响。方法:通过激活多肽活化细胞膜表面的蛋白酶活化受体2,检测对结肠癌细胞与FN和Matrigel粘附能力的影响。建立蛋白酶活化受2敲降的结肠癌细胞,检测对结肠癌细胞与基质粘附能力的影响。结果:蛋白酶活化受体2激活,能够促进结肠癌细胞与基质的粘附能力;而蛋白酶活化受体敲降则抑制粘附。结论:蛋白酶活化受体2参与对结肠癌细胞粘附能力的调控,其激活能够促进结肠癌细胞与基质的粘附能力。     

Fibronectin and culture temperature modulate the efficacy of an avidin-biotin binding system for chondrocyte adhesion and growth on biodegradable polymers

Cell adhesion to a scaffold is a prerequisite for tissue engineering. Many studies have been focused on enhancing cell adhesion to synthetic materials that are used for scaffold fabrication. Previously, we showed that immobilization of biotin molecules to chondrocyte surfaces enhanced cell adhesion to avidin-coated biodegradable polymers such as poly-L-lactic acid, poly-D,L-lactic acid and polycaprolactone. However, the endocytosis of cell membrane biotin molecules decreases binding strength between biotinylated-chondrocytes (B-chondrocytes) and avidin-coated substrata, and therefore decreases cell spreading and discourages long-term chondrocytes culture. In this study, we proposed two strategies to solve the shortcoming of the avidin-biotin binding system. First, the avidin-biotin binding system is combined with the intrinsic integrin-dependent adhesion systems in order to enhance long-term cell culture. Second, the incubation temperature is lowered in order to slow down the endocytosis process. We found that the avidin-biotin binding system in combination with FN-integrin binding system enhanced cell adhesion, cell spreading and cell growth. Decrease of cell culture temperature to 4 degrees C enhanced the adhesion of B-chondrocytes to the avidin-coated surfaces, but decreased cell viability and proliferation, compared to culture temperature of 37 degrees C. Whether there is an optimal seeding temperature between 4 and 37 degrees C for both adhesion and proliferation of B-chondrocytes needs further investigation. Our results indicated that modulation of the adhesion conditions could further enhance the efficacy of the avidin-biotin binding system in mediating cell adhesion, and subsequent tissue culture.     

Mechanism of Xenopus cranial neural crest cell migration

This Review focuses on recent advances in the field of cranial neural crest cell migration in Xenopus laevis with specific emphasis on cell adhesion and the regulation of cell migration. Our goal is to combine the understanding of cell adhesion to the extracellular matrix with the regulation of cell-cell adhesion and the involvement of the planar cell polarity signaling-pathway in guiding the migration of cranial neural crest cells during embryogenesis.Key words: neural crest, cell migration, extracellular matrix, cell adhesion, Wnt, planar cell polarity     

细胞表面糖链末端唾液酸和岩藻糖和某些细胞生物学行为的关系

用神经氨酸酶和α-L-岩藻糖苷酶分别切除人肝癌细胞株7721细胞表面糖链中的末端唾液酸（SA）和岩藻糖（Fuc)残基来研究表面聚糖结构和某些细胞生物学行为之间的关系。选择细胞对纤连蛋白（Fn）,层黏蛋白（Ln)和人脐静脉内皮细胞（HUVEC）的黏附能力,细胞趋化性迁移以及趋化性侵袭作为细胞行为的指标。结果表明：表面人脐静脉内皮细胞（HUVEC）的黏附能力,细胞趋化性迁移以及趋化性侵袭作为细胞行为的指标。结果表明：表面糖链末端SA对细胞黏附至Fn并不必需,对细胞黏附至Ln和细胞的趋化性侵袭却至为重要,而对细胞黏附至HUVEC以及趋化性迁移则为关键性残基。与SA相比,Fuc可能参与细胞Fn,Ln和HUVEC的黏附,但对趋化性迁移以及趋化性侵袭并不重要。细胞对HUVEC的黏附以及趋化性迁移和侵袭可被唾液酸化Lewis X(SLe^x)单抗抑制,但不被未唾液酸化的Lewis X(Le^x)单抗抑制,这一结果支持SA在上述三种细胞过程中Fuc残基重要。