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PubMed2004

The chemokine network in cancer--much more than directing cell movement.

Cytokine and chemokine gradients are central to the directed movement of cells in both homeostatic and pathological processes. Most cancers have a complex chemokine network which can influence immune responses to the tumor, direct the extent and cellular composition of the leukocyte infiltrate and also play a role in angiogenesis. Tumor cells can also hijack the chemokine system and gain expression of certain chemokine receptors and respond to specific chemokine gradients. Chemokine receptor expression and activation on malignant cells may be central to the growth, survival and migration of cancer cells from the primary tumor. Chemokine receptors, both CC and CXC have been detected on malignant cells and the relevant ligands are sometimes expressed at the tumor site and at sites of tumor spread, suggesting a role for the chemokine family in malignant growth and metastasis.

Animals↗

PubMed1989

Studies of bronchoalveolar lavage cells and fluids in pulmonary sarcoidosis. II. Enhanced capacity of bronchoalveolar lavage fluids from patients with pulmonary sarcoidosis to induce cell movement in vitro.

The ability to increase the motility of endothelial cells in vitro is a property common to most if not all angiogenesis-inducing factors. Because bronchoalveolar lavage (BAL) cells from patients with pulmonary sarcoidosis have an enhanced capacity to induce neovascularization, the BAL fluids from these patients were assessed for their effect on human and murine endothelial cells and fibroblasts obtained from a variety of tissue sources. A recently developed computer-assisted image analysis system was used to determine the extent and pattern of cell migration in a microwell screening assay. Data were obtained for BAL fluids from 10 patients with pulmonary sarcoidosis and from five normal volunteers. BAL supernatants from patients with active sarcoidosis showed an enhanced (2- to 8-fold) capacity to induce chemokinesis of both endothelial cells and fibroblasts, as measured by increased area of migration and polarized cell movement. There was a marked heterogeneity in the motility of cells from different organ origins, but enhanced cell movement was observed with both endothelial cells and fibroblasts. In contrast, BAL fluids from normal and sarcoid patients were similar in their effect on muscle cells and urothelial cells, whereas pericytes, which responded to BAL fluids from normal subjects or patients with nongranulomatous pulmonary disease, were inhibited by BAL fluids from patients with pulmonary sarcoidosis. The induction of endothelial cell movement in vitro induced by individual supernatants generally correlated with the capacity of BAL cells from these patients to induce angiogenesis in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1999

Interactive image processing system for assessment of cell movement.

The study of cancer cell motility is considered to be important in understanding cancer metastasis. The movement behaviour of cells within clustered cell colonies is of particular interest. Changes in cell movement, area and velocity can be an indicator of cell spreading. The aim of the study is to develop and apply a computerised interactive image processing system to quantify the movement of cells within cell clusters. A semi-automatic boundary description method based on two-dimensional rendering is devised. The system is later combined with image-processing methods that facilitate the relocation of the cell boundary over time; this forms a new approach to assessing cell movement. These methods are incorporated into a software system, enabling an interactive procedure to define and monitor the movement of single cells in cell clusters from digitised microscope images. Validation of the method shows a maximum error of 10% in defining the area through a cubic spline interpolation. The system is applied to analyse the movement and area of HT115 human colon cancer cells. The system provides tools for the analysis of movement, area and velocity of single cells in cancer cell colonies and may thus be of value in further understanding cancer cell motility.

Cell Movement↗

PubMed2002

Collective cell movement in primary melanoma explants: plasticity of cell-cell interaction, beta1-integrin function, and migration strategies.

Collective cell movement represents an efficient dissemination strategy in neoplastic epithelial and mesenchymal cancer. In primary melanoma explants cultured in three-dimensional collagen lattices, invasive migration of multicellular clusters was dependent on the function of beta1 integrins, as shown by preferential beta1-integrin expression and clustering in a subset of promigratory cells at the leading edge ("guiding cells") and the abrogation of multicellular migration by adhesion-perturbing anti-beta1-integrin antibody. Interference with beta1-integrin function induced complex changes in cluster polarity and cohesion, including development of two or several opposing leading edges, cluster disruption, and the detachment of individual cells followed by beta1-integrin-independent "amoeboid" crawling and dissemination. The conversion from beta1-integrin-dependent collective movement to beta1-integrin-independent single-cell motility suggests efficient cellular and molecular plasticity in tumor cell migration strategies.

Cell Adhesion↗

PubMed1992

On the convergent cell movements of gastrulation in Fundulus.

Mainly because of its transparency, the Fundulus gastrula constitutes ideal material for direct study of morphogenetic cell movements in vivo. Marking studies show that deep cells of the germ ring converge toward and enter the embryonic shield, where they undergo extension. Those close to the shield move faster. Analysis of videotapes reveals that all deep cells of the dorsal germ ring move toward the shield. But none moves in a direct line. All meander considerably. Germ ring cells nearer the shield move toward it at a higher net rate than those farther away because they meander less. This suggests that exogenous factors promote their directionality. Cells in the prospective yolk sac adjacent to the germ ring also show net convergence, but they meander more. Directional forces are apparently stronger in the germ ring. Converging deep cells move both by filolamellipodia and, less frequently, by blebs. However, there is very little individual cell movement; all cells are almost always in adhesive contact with other cells in moving cell clusters. Clusters vary constantly in size, continually aggregating with other cells and other clusters and splitting. Filolamellipodial cells show contact inhibition of cell movement. Nevertheless, they move and do so directionally, presumably in part because, as members of cell clusters, much of their movement is passive. They also show intercalation or invasive activity, but, consistent with their contact-inhibiting properties, only when neighboring cells separate and provide free space. Cells moving by blebbing locomotion are non-contact inhibiting and intercalate readily. Cell division continues during convergence. Although this temporarily arrests their movement, the daughter cells soon join in the mass convergent movement.

Animals↗

PubMed1990

An autoradiography study of myogenic cell movement in avian limb buds following heterospecific and homospecific transplantation.

Species specificity and the use of quail cells as a marker in the study of myogenic cell movement in the developing avian limb was investigated. In order to establish whether or not observed myogenic cell movement in quail/chick limb transplantation experiments might be an artefact produced by cellular interaction between these cell types a series of homospecific and heterospecific transplantations was performed. Chick wing fragments (staged 20-25 H.H.) were labelled with tritiated thymidine and inserted into unlabelled chick wing bud (homospecific) in ovo. In addition, quail wing fragments were also labelled with tritiated thymidine and transplanted in the same manner into chick (heterospecific), so that the effectiveness of tritium as a marker could be assessed. After 4 days post-incubation, myogenic cell movement was detected in eight out of the ten homospecific transplantations performed. Myogenic cell movement in avian limbs is therefore not produced by interaction between chick and quail cells, as migration was also detected in the chick/chick transplants. Nonetheless, heterospecific transplantation results revealed that autoradiographic methods failed to reveal completely the true extent to which myogenic cell movement occurred, because tritiated thymidine was subject to dilution.

Animals↗

PubMed1988

Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis.

We have found that the spontaneous migration of bovine aortic endothelial cells from the edge of a denuded area in a confluent monolayer is dependent upon the release of endogenous basic fibroblast growth factor (bFGF). Cell movement is blocked by purified polyclonal rabbit IgG to bFGF as well as affinity purified anti-bFGF IgG and anti-bFGF F(ab')2 fragments. The inhibitory effect of the immunoglobulins is dependent upon antibody concentration, is reversible, is overcome by the addition of recombinant bFGF, and is removed by affinity chromatography of the antiserum through a column of bFGF-Sepharose. Cell movement is also reversibly inhibited by the addition of protamine sulfate and suramin; two agents reported to block bFGF binding to its receptor. The addition of recombinant bFGF to wounded monolayers accelerates the movement of cells into the denuded area. Transforming growth factor beta which has been shown to antagonize several other effects of bFGF also inhibits cell movement. The anti-bFGF IgG prevents the movement of bovine capillary endothelial cells, BHK-21, NIH 3T3, and human skin fibroblasts into a denuded area. Antibodies to bFGF, as well as suramin and protamine sulfate also suppress the basal levels of plasminogen activator and DNA synthesis in bovine aortic endothelial cells.

Animals↗

PubMed1996

Intraperitoneal cell movement during abdominal carbon dioxide insufflation and laparoscopy. An in vivo model.

PURPOSE: Possible mechanisms of movement of malignant cells within the peritoneal cavity during CO2 insufflation and laparoscopy involve direct transfer via laparoscopic instruments or dispersion of cells by CO2 or water vapor. An in vivo model has been developed to study these mechanisms. METHODS: Laparoscopy was performed on an animal model (domestic white pig). Cells derived from colorectal cancer cell line Lim 1215 were injected to lie free within the peritoneal cavity. A polycarbonate filter system with a 5-micron pore diameter was used to examine CO2 expelled from the peritoneal cavity, during laparoscopy and manipulation of abdominal viscera, for malignant cells. Laparoscopic instruments and laparoscopic ports were washed independently, and fluid was centrifuged and examined for malignant cells. RESULTS: Malignant cells were identified on 1 of 30 filters used to examine exhaust carbon dioxide. Malignant cells also were identified from 2 of 10 washings from laparoscopic ports and from 4 of 10 washings of laparoscopic instruments. CONCLUSIONS: These results suggest that movement of cells throughout the peritoneal cavity during laparoscopy is via contaminated instruments, but local cell movement by dispersion possibly within water vapor from the port may also occur.

Animals↗

PubMed2004

The C terminus of the movement protein of Brome mosaic virus controls the requirement for coat protein in cell-to-cell movement and plays a role in long-distance movement.

The 3a movement protein (MP) plays a central role in the movement of Brome mosaic virus (BMV). To identify the functional regions in BMV MP, 24 alanine-scanning (AS) MP mutants of BMV were constructed. Infectivity of the AS mutants in the host plant Chenopodium quinoa showed that the central region of BMV MP is important for viral movement and both termini of BMV MP have effects on the development of systemic symptoms. A green-fluorescent-protein-expressing RNA3-based BMV vector containing a 2A sequence from Foot-and-mouth disease virus was also constructed. Using this vector, two AS mutants that showed more efficient cell-to-cell movement than wild-type BMV were identified. The MPs of these two AS mutants, which have mutations at their C termini, mediated cell-to-cell movement independently of coat protein (CP), unlike wild-type BMV MP. Furthermore, a BMV mutant with a truncation in the C-terminal 42 amino acids of MP was also able to move from cell to cell without CP, but did not move systemically, even in the presence of CP. These results and an encapsidation analysis suggest that the C terminus of BMV MP is involved in the requirement for CP in cell-to-cell movement and plays a role in long-distance movement. Furthermore, the ability to spread locally and form virions is not sufficient for the long-distance movement of BMV. The roles of MP and CP in BMV movement are discussed.

Bromovirus↗

PubMed1980

Formation of protrusions of the cell surface during tissue cell movement.

The various forms of protrusions of the surface of tissue cells are described in relation to their role in cell locomotion. It is proposed that these cells have enough cell surface (plasma membrane and linked microfilamentous cortical cytoplasm) at any given moment during interphase of the cell cycle to satisfy their need for the local increases in cell surface area that accompany protrusive activity. Thus, a local increase in protrusive activity in one region of the cell surface would be accompanied by a corresponding decrease elsewhere; that is, formation of new protrusions would require retraction of other protrusions already present and the area retracted would be equivalent to the area protruded. Evidence marshaled in support of this hypothesis includes disappearance of microvilli and other microprotrusions during cell spreading, increase in protrusive activity of uncontacted regions of the cell surface during contact inhibition of cell movement, antagonism between blebbing and spreading, accelerated protrusive activity at the leading edge upon abrupt retraction of the trailing edge, surface flow during bleb formation, and antagonism between various protrusive activities associated with cell movement and cytokinesis. Finally, the relevance of these findings to two important developmental problems is explored: the commencement of gastrulation and directional cell movements during morphogenesis.

Animals↗

PubMed1994

Epidermal growth factor receptor-mediated cell motility: phospholipase C activity is required, but mitogen-activated protein kinase activity is not sufficient for induced cell movement.

We recently have demonstrated that EGF receptor (EGFR)-induced cell motility requires receptor kinase activity and autophosphorylation (P. Chen, K. Gupta, and A. Wells. 1994. J. Cell Biol. 124:547-555). This suggests that the immediate downstream effector molecule contains a src homology-2 domain. Phospholipase C gamma (PLC gamma) is among the candidate transducers of this signal because of its potential roles in modulating cytoskeletal dynamics. We utilized signaling-restricted EGFR mutants expressed in receptor devoid NR6 cells to determine if PLC activation is necessary for EGFR-mediated cell movement. Exposure to EGF (25 nM) augmented PLC activity in all five EGFR mutant cell lines which also responded by increased cell movement. Basal phosphoinositide turnover was not affected by EGF in the lines which do not present the enhanced motility response. The correlation between EGFR-mediated cell motility and PLC activity suggested, but did not prove, a causal link. A specific inhibitor of PLC, U73122 (1 microM) diminished both the EGF-induced motility and PLC responses, while its inactive analogue U73343 had no effect on these responses. Both the PLC and motility responses were decreased by expression of a dominant-negative PLC gamma-1 fragment in EGF-responsive infectant lines. Lastly, anti-sense oligonucleotides (20 microM) to PLC gamma-1 reduced both responses in NR6 cells expressing wild-type EGFR. These findings strongly support PLC gamma as the immediate post receptor effector in this motogenic pathway. We have demonstrated previously that EGFR-mediated cell motility and mitogenic signaling pathways are separable. The point of divergence is undefined. All kinase-active EGFR mutants induced the mitogenic response while only those which are autophosphorylated induced PLC activity. U73122 did not affect EGF-induced thymidine incorporation in these motility-responsive infectant cell lines. In addition, the dominant-negative PLC gamma-1 fragment did not diminish EGF-induced thymidine incorporation. All kinase active EGFR stimulated mitogen-activated protein (MAP) kinase activity, regardless of whether the receptors induced cell movement; this EGF-induced MAP kinase activity was not affected by U73122 at concentrations that depressed the motility response. Thus, the signaling pathways which lead to motility and cell proliferation diverge at the immediate post-receptor stage, and we suggest that this is accomplished by differential activation of effector molecules.

Base Sequence↗

PubMed1998

Cell-to-cell movement of turnip crinkle virus is controlled by two small open reading frames that function in trans.

Previous studies on turnip crinkle virus (TCV) have suggested that the two small, centrally located ORFs, conserved in all Carmoviruses, are both required for cell-to-cell movement (Hacker et al., 1992). We now demonstrate that the cell-to-cell movement of TCV is mediated by in trans complementation of the two proteins. First, both of the putative movement proteins (MPs p8 and p9) were shown to be translated in vitro from transcripts representing the 1.7-kb subgenomic RNA. Western blot analysis, using antisera prepared against GST fusion proteins of both genes, was then used to show that the p8 but not the p9 protein accumulated to detectable levels in particulate fractions of infected cells. Cell-to-cell movement of various MP mutants in Arabidopsis was evaluated by in situ hybridization of inoculated leaves. Changes in either of the two MP genes resulted in failure of the mutants to move cell-to-cell. Coat protein was found to be unnecessary for cell-to-cell movement. Complementation of cell-to-cell movement by co-inoculating p8-defective mutants with a p9-defective mutant resulted in delayed systemic infection. In contrast, efficient cell-to-cell movement was achieved when the MP mutants were inoculated into transgenic plants expressing the corresponding functional gene(s). These experiments provide further evidence that both MP genes encoded by Carmoviruses must function in trans in the same cell in order to mediate cell-to-cell movement.

Arabidopsis↗

PubMed1996

FGF-2 influences cell movements and gene expression during limb development.

FGF-2 is proposed to be an important ectodermal signal directing limb outgrowth and patterning. Consistent with this hypothesis we show that ectopic application of FGF-2 can maintain the apical ectodermal ridge (AER)-dependent expression of Sonic hedgehog (Shh), and AER-dependent zone of polarizing activity (ZPA) signaling. We also find that ectopic FGF-2 applied to the posterior wing bud caused a dramatic change in the morphology of the limb bud, and results in limbs that display a reduction in the length of individual skeletal elements and loss of digits. Associated with these morphological changes was an FGF-2-stimulated expansion and bifurcation of the expression domains of two posteriorly expressed genes, Shh and HoxD13. Applying FGF-2 at a central or anterior location in the limb bud did not alter the Shh expression domain or cause digit loss. To test whether ectopic application of FGF-2 into the posterior limb bud was influencing the movement of limb bud cells, we used the lipophilic dye DiI to map the behavior of posterior cells in response to FGF-2. In response to FGF-2 posterior limb bud cells move in both a proximal and a distal direction, causing the initially labeled cell population to bifurcate into two distinct domains. Our data suggest that FGF-2 is influencing limb outgrowth by modifying cell movements and subsequent position-specific cell-cell interactions that are important for limb morphogenesis.

Animals↗

PubMed1994

Patterns of cell movement within the Dictyostelium slug revealed by cell type-specific, surface labeling of living cells.

There are cells scattered in the rear, prespore region of the Dictyostelium slug that share many of the properties of the prestalk cells and that are therefore called anterior-like cells (ALCs). By placing the gene encoding a cell surface protein under the control of an ALC-specific promoter and immunologically labeling the living cells, we analyze the movement of ALCs within the slug. There is a posterior to anterior cellular flow, and the ALCs change their movement pattern as they enter the prestalk zone. Prestalk cells are periodically shed from the migrating slug. They must be replaced if the correct ratio of prestalk to prespore cells is to be maintained, and we present evidence for the transdifferentiation of prespore into prestalk cells, with ALCs functioning as intermediates in the transition. The slug has, therefore, a surprisingly dynamic structure, both with respect to cellular differentiation and cell movement.

Amino Acid Sequence↗

PubMed1999

In vivo real-time analysis of intraperitoneal radiolabeled tumor cell movement during laparoscopy.

PURPOSE: A porcine model has been developed to allow the real-time imaging of radiolabeled tumor cell movement throughout the peritoneal cavity, both at rest and during carbon dioxide insufflation. METHODS: Fifteen 30-kg domestic white female pigs were used. Under anesthesia, 15 to 20 million radiolabeled human colorectal tumor cells (LIM1215) were introduced into the peritoneal cavity under laparoscopic vision into the pelvis. Radiolabeled tumor cell movement was examined by using a 25-cm-diameter, low-energy mobile gamma camera with high resolution collimator. Tumor cell movement and distribution during two hours without insufflation was examined in four pigs. Then tumor cell movement and distribution during two hours with CO2 insufflation was examined in four pigs. In a further four pigs, tumor cells were then mixed with blood and injected into the peritoneal cavity and the effect of no insufflation vs. insufflation was noted. A further three pigs were examined with manipulation of the intra-abdominal contents after injection of LIM1215 cells into the peritoneal cavity. Venting insufflating CO2 was filtered for tumor cells. RESULTS: Widespread intraperitoneal distribution of tumor cells from the pelvis was identified both with CO2 insufflation of the peritoneal cavity and without insufflation. Tumor cells dispersed throughout the peritoneal cavity at a slower rate without carbon dioxide insufflation. There was a differential rate of tumor cell movement to the left upper quadrant and right upper quadrant with insufflation and without insufflation. Blood within the peritoneal cavity and an extended contact of the laparoscopic trocars with the peritoneal cavity in this setting increased contamination of the trocars and trocar sites with tumor cells. Tumor cells were identified on laparoscopic instruments in all experiments. No evidence of aerosolization of tumor cells was found. CONCLUSION: Tumor cells move throughout the peritoneal cavity both at rest and during CO2 insufflation. The pattern of tumor cell dispersion differs with CO2 insufflation. The presence of blood and extended contact of trocars with peritoneal contents are a major factor in trocar and trocar site tumor cell contamination.

Animals↗

PubMed1997

A Model for Cell Movement During Dictyostelium Mound Formation

Dictyostelium development is based on cell-cell communication by propagating cAMP signals and cell movement in response to these signals. In this paper we present a model describing wave propagation and cell movement during the early stages of Dictyostelium development, i.e. aggregation and mound formation. We model cells as distinct units whose cAMP relay system is described by the Martiel-Goldbeter model. To describe cell movement we single out three components: chemotactic motion, random motion and motion due to pressure between cells. This pressure result in cells crawling on top of each other and therefore to the extension of the aggregate into the third dimension. Using this model we are able to describe aggregation up to the mound stage. The cells in the mound move in a rotational fashion and their movement is directed by the counter-rotating spiral of the chemo-attractant cAMP. Furthermore, we show that the presence of two subpopulations with different inherent chemotactic velocities can lead to cell sorting in the mound. The fast moving cells collect into the centre while the slow cells occupy the rest of the mound. This model allows the direct comparison of the properties of the cAMP waves properties and movement behavior of individual cells with experimental data. Thereby it allows a critical test of our understanding of the basic cellular principles involved in the morphogenesis of a simple eukaryote.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited

Journal Article↗

PubMed1993

The roles of the red clover necrotic mosaic virus capsid and cell-to-cell movement proteins in systemic infection.

The red clover necrotic mosaic dianthovirus (RCNMV) genome is split between two single-stranded RNA species termed RNA-1 and RNA-2. RNA-2 is required for infection of whole plants but is dispensable for infection and virion formation in protoplasts. We have used full-length cDNA clones of RNA-1 and -2 from which infectious in vitro transcripts can be derived to construct a number of mutations in the RNA-1 encoded capsid protein and the RNA-2 encoded cell-to-cell movement protein genes. The capsid protein and the RNA sequence encoding the capsid protein were dispensable for infection of the inoculated leaves of Nicotiana benthamiana and N. clevelandii at both 15 and 25 degrees. In addition, capsid protein was not necessary for systemic infection of N. benthamiana at 15 degrees. As many as 39 amino acid residues could be deleted from the carboxyl-terminus of the RNA-2 encoded 35-kDa cell-to-cell movement protein without loss of or reduction in the rate of cell-to-cell movement or systemic infection. However, larger deletions within the cell-to-cell movement protein gene prevented cell-to-cell movement and systemic infection of N. benthamiana. These data suggest that the spread of RCNMV in a systemic host is a combination of two distinct events: cell-to-cell movement and long distance transport. We conclude that the RCNMV 35-kDa movement protein is required for cell-to-cell movement, whereas the capsid protein is not necessary for cell-to-cell movement and, depending on host genotype and environmental factors, may or may not be required for long distance transport.

Base Sequence↗

PubMed2004

Rice dwarf phytoreovirus segment S6-encoded nonstructural protein has a cell-to-cell movement function.

Rice dwarf virus (RDV) is a member of the genus Phytoreovirus, which is composed of viruses with segmented double-stranded RNA genomes. Proteins that support the intercellular movement of these viruses in the host have not been identified. Microprojectile bombardment was used to determine which open reading frames (ORFs) support intercellular movement of a heterologous virus. A plasmid containing an infectious clone of Potato virus X (PVX) defective in cell-to-cell movement and expressing either beta-glucuronidase or green fluorescent protein (GFP) was used for cobombardment with plasmids containing ORFs from RDV gene segments S1 through S12 onto leaves of Nicotiana benthamiana. Cell-to-cell movement of the movement-defective PVX was restored by cobombardment with a plasmid containing S6. In the absence of S6, no other gene segment supported movement. Identical results were obtained with Nicotiana tabacum, a host that allows fewer viruses to infect and spread within its tissue. S6 supported the cell-to-cell movement of the movement-defective PVX in sink and source leaves of N. benthamiana. A mutant S6 lacking the translation start codon did not complement the cell-to-cell movement of the movement-defective PVX. An S6 protein product (Pns6)-enhanced GFP fusion was observed near or within cell walls of epidermal cells from N. tabacum. By immunocytochemistry, unfused Pns6 was localized to plasmodesmata in rice leaves infected with RDV. S6 thus encodes a protein with characteristics identical to those of other viral proteins required for the cell-to-cell movement of their genome and therefore is likely required for the cell-to-cell movement of RDV.

Open Reading Frames↗