Localization of urokinase to focal adhesions by human fibrosarcoma cells synthesizing recombinant vitronectin

1996 ◽  
Vol 74 (6) ◽  
pp. 899-910 ◽  
Author(s):  
Sarah A. Wilcox ◽  
Thomas Reho ◽  
Eiman Tominna-Sebald ◽  
Paula J. Mckeown-Longo ◽  
Paul J. Higgins
Keyword(s):  
Cell Migration ◽  
Cell Surface ◽  
Intracellular Signaling ◽  
De Novo ◽  
Focal Adhesions ◽  
Migration And Invasion ◽  
Cdna Insert ◽  
Transfected Cells ◽  
Matrix Deposition ◽  
Fibrosarcoma Cells

Cell surface plasminogen activators have been proposed to participate in cell migration and invasion by activating both intracellular signaling pathways and extracellular proteolysis. Urokinase-type plasminogen activator (uPA) is secreted from many cell types and localizes to focal contact areas when cells are seeded onto the plasma protein vitronectin. Induction of vitronectin synthesis during migration of neural crest cells and growth of certain tumors suggests that the de novo synthesis and deposition of vitronectin into the tissue matrix may remodel the matrix to provide an environment suitable for cell migration and (or) tumor invasion. To investigate the effects of vitronectin secretion and matrix deposition on the localization and activity of cell-associated uPA, HT-1080 fibrosarcoma cells were transfected with the Rc/CMV expression vector containing a vitronectin cDNA insert and stable cell lines expressing vitronectin were selected. Vitronectin-secreting cells were allowed to attach and spread on collagen- and fibronectin-coated substrates. Within 6 h, vitronectin was detected on the substrate; vitronectin synthesis was accompanied by the clustering of both the αvβ5 vitronectin receptor and uPA into vinculin-containing focal adhesions. Although mock transfected cells formed small focal adhesions on both collagen and fibronectin, no co-localization of uPA or αvβ5 to focal adhesions was evident in these cells. Vitronectin-secreting cells also exhibited decreased levels of plasminogen activation and increased levels of cell adhesion as compared with the mock transfected cells. These data demonstrate that the synthesis of vitronectin and its matrix association by transfected HT-1080 fibrosarcoma cells results in localization of uPA to oαvβ5 containing focal adhesions, decreased cell surface uPA activity, and an increase in cell adhesion.Key words: urokinase, vitronectin, focal adhesions, αvβ5 integrin.

scholarly journals Rab40/Cullin5 complex regulates EPLIN and actin cytoskeleton dynamics during cell migration and invasion

2021 ◽  
Author(s):  
Erik S Linklater ◽  
Emily Duncan ◽  
Ke Jun Han ◽  
Algirdas Kaupinis ◽  
Mindaugas Valius ◽  
...  
Keyword(s):  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Leading Edge ◽  
Stress Fibers ◽  
Actin Dynamics ◽  
Migration And Invasion ◽  
Matrix Remodeling ◽  
Cell Migration And Invasion

Rab40b is a SOCS box containing protein that regulates the secretion of MMPs to facilitate extracellular matrix remodeling during cell migration. Here we show that Rab40b interacts with Cullin5 via the Rab40b SOCS domain. We demonstrate that loss of Rab40b/Cullin5 binding decreases cell motility and invasive potential, and show that defective cell migration and invasion stem from alteration to the actin cytoskeleton, leading to decreased invadopodia formation, decreased actin dynamics at the leading edge, and an increase in stress fibers. We also show that these stress fibers anchor at less dynamic, more stable focal adhesions. Mechanistically, changes in the cytoskeleton and focal adhesion dynamics are mediated in part by EPLIN, which we demonstrate to be a binding partner of Rab40b and a target for Rab40b/Cullin5 dependent localized ubiquitylation and degradation. Thus, we propose a model where the Rab40b/Cullin5 dependent ubiquitylation regulates EPLIN localization to promote cell migration and invasion by altering focal adhesion and cytoskeletal dynamics.

scholarly journals Requirements for sulfate transport and the diastrophic dysplasia sulfate transporter in fibronectin matrix assembly

2007 ◽  
Vol 179 (5) ◽  
pp. 999-1009 ◽  
Author(s):  
Leontine L. Galante ◽  
Jean E. Schwarzbauer
Keyword(s):  
Cell Surface ◽  
Sulfate Transporter ◽  
Matrix Assembly ◽  
Diastrophic Dysplasia ◽  
Matrix Deposition ◽  
Fibrosarcoma Cells ◽  
Extracellular Matrix Deposition ◽  
Necessary And Sufficient ◽  
Rna Knockdown ◽  
Interfering Rna

Diastrophic dysplasia sulfate transporter (DTDST) is a sulfate/chloride antiporter whose function is impaired in several human chondrodysplasias. We show that DTDST is upregulated by dexamethasone stimulation of HT1080 fibrosarcoma cells and is required for fibronectin (FN) extracellular matrix deposition by these cells. DTDST imports sulfate for the modification of glycosaminoglycans. We find that N-sulfation of these chains is important for FN matrix assembly and that sulfation of cell surface proteoglycans is reduced in the absence of DTDST. Of the candidate HT1080 cell surface proteoglycans, only loss of syndecan-2 compromises FN assembly, as shown by syndecan-2 small interfering RNA knockdown. DTDST is both necessary and sufficient to induce FN matrix assembly in HT1080 cells. Knockdown of DTDST ablates FN matrix, whereas its overexpression increases assembly without dexamethasone stimulation. These results identify a previously unrecognized regulatory pathway for matrix assembly via modulation of a sulfate transporter and proteoglycan sulfation. These data raise the possibility that FN assembly defects contribute to chondrodysplasias.

The interaction of enolase-1 with caveolae-associated proteins regulates its subcellular localization

2014 ◽  
Vol 460 (2) ◽  
pp. 295-307 ◽  
Author(s):  
Dariusz Zakrzewicz ◽  
Miroslava Didiasova ◽  
Anna Zakrzewicz ◽  
Andreas C. Hocke ◽  
Florian Uhle ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Surface ◽  
Subcellular Localization ◽  
Migration And Invasion ◽  
Caveolin 1 ◽  
Annexin 2 ◽  
Associated Proteins ◽  
Surface Localization ◽  
Dependent Cell ◽  
Cell Surface Localization

We found that ENO-1 localizes to caveolae and interacts with caveolin-1 and annexin 2. The association of ENO-1 with caveolar proteins and localization within caveolae are required for ENO-1 cell surface localization and ENO-1-dependent cell migration and invasion.

scholarly journals Intracellular nanovesicles mediate integrin trafficking during cell migration

2020 ◽  
Author(s):  
Gabrielle Larocque ◽  
Penelope J. La-Borde ◽  
Beverley J. Wilson ◽  
Nicholas I. Clarke ◽  
Daniel J. Moore ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Surface ◽  
Membrane Trafficking ◽  
Surface Proteins ◽  
Migration And Invasion ◽  
Rab Gtpase ◽  
Surface Receptors ◽  
C Terminus ◽  
Transport Vesicle ◽  
Important Regulator

Membrane traffic is an important regulator of cell migration through the endocytosis and recycling of cell surface receptors such as integrin heterodimers. Intracellular nanovesicles (INVs), are a recently identified class of transport vesicle that are involved in multiple membrane trafficking steps including the recycling pathway. The only known marker for INVs is Tumor Protein D54 (TPD54/TPD52L2), a member of the TPD52-like protein family. Overexpression of TPD52-like family proteins in cancer has been linked to poor prognosis and an aggressive metastatic phenotype which suggests cell migration may be altered under these conditions. Here we show that TPD54 associates with INVs by directly binding high curvature membrane via a conserved positively charged motif in its C-terminus. We describe how other members of the TPD52-like family are also associated with INVs and we document the Rab GTPase complement of all INVs. Depletion of TPD52-like proteins inhibits cell migration and invasion; and we show that this is likely due to altered integrin recycling. Our study highlights the involvement of INVs in the trafficking of cell surface proteins to generate biologically important outputs in health and disease.

scholarly journals Leptin induces cell migration and invasion in a FAK-Src-dependent manner in breast cancer cells

2019 ◽  
Vol 8 (11) ◽  
pp. 1539-1552 ◽  
Author(s):  
Juan Carlos Juárez-Cruz ◽  
Miriam Daniela Zuñiga-Eulogio ◽  
Monserrat Olea-Flores ◽  
Eduardo Castañeda-Saucedo ◽  
Miguel Ángel Mendoza-Catalán ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Migration ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cells ◽  
Focal Adhesions ◽  
Migration And Invasion ◽  
Breast Cancer Cell Lines ◽  
Dependent Manner

Breast cancer is the most common invasive neoplasia, and the second leading cause of the cancer deaths in women worldwide. Mammary tumorigenesis is severely linked to obesity, one potential connection is leptin. Leptin is a hormone secreted by adipocytes, which contributes to the progression of breast cancer. Cell migration, metalloproteases secretion, and invasion are cellular processes associated with various stages of metastasis. These processes are regulated by the kinases FAK and Src. In this study, we utilized the breast cancer cell lines MCF7 and MDA-MB-231 to determine the effect of leptin on FAK and Src kinases activation, cell migration, metalloprotease secretion, and invasion. We found that leptin activates FAK and Src and induces the localization of FAK to the focal adhesions. Interestingly, leptin promotes the activation of FAK through a Src- and STAT3-dependent canonical pathway. Specific inhibitors of FAK, Src and STAT3 showed that the effect exerted by leptin in cell migration in breast cancer cells is dependent on these proteins. Moreover, we established that leptin promotes the secretion of the extracellular matrix remodelers, MMP-2 and MMP-9 and invasion in a FAK and Src-dependent manner. Our findings strongly suggest that leptin promotes the development of a more aggressive invasive phenotype in mammary cancer cells.

scholarly journals Hypoxia-inducible Factor Regulates αvβ3 Integrin Cell Surface Expression

2005 ◽  
Vol 16 (4) ◽  
pp. 1901-1912 ◽  
Author(s):  
Karen D. Cowden Dahl ◽  
Sarah E. Robertson ◽  
Valerie M. Weaver ◽  
M. Celeste Simon
Keyword(s):  
Cell Surface ◽  
Cell Fate ◽  
Hypoxia Inducible Factor ◽  
Focal Adhesions ◽  
Surface Expression ◽  
Melanoma Cells ◽  
Cell Surface Expression ◽  
Migration And Invasion ◽  
Αvβ3 Integrin ◽  
And Migration

Hypoxia-inducible factor (HIF)-deficient placentas exhibit a number of defects, including changes in cell fate adoption, lack of fetal angiogenesis, hypocellularity, and poor invasion into maternal tissue. HIF is a heterodimeric transcription factor consisting of α and β aryl hydrocarbon receptor nuclear translocator or ARNT) subunits. We used undifferentiated trophoblast stem (TS) cells to characterize HIF-dependent adhesion, migration, and invasion. Arnt-/- and Hifα-/- TS cells exhibit reduced adhesion and migration toward vitronectin compared with wild-type cells. Furthermore, this defect is associated with decreased cell surface expression of integrin αvβ3 and significantly decreased expression of this integrin in focal adhesions. Because of the importance of adhesion and migration in tumor progression (in addition to placental development), we examined the affect of culturing B16F0 melanoma cells in 1.5% oxygen (O2). Culturing B16F0 melanoma cells at 1.5% O2 resulted in increased αvβ3 integrin surface expression and increased adhesion to and migration toward vitronectin. Together, these data suggest that HIF and O2 tension influence placental invasion and tumor migration by increasing cell surface expression of αvβ3 integrin.

Elevated O-GlcNAcylation Promotes Malignant Phenotypes of Hypopharyngeal Squamous Cell Carcinoma by Stabilizing Nrf2 through Regulation of the PI3K/Akt Pathway

2020 ◽  
Vol 20 (16) ◽  
pp. 1933-1942
Author(s):  
Wencheng Dai ◽  
Xiaoxia Jin ◽  
Bin Jiang ◽  
Weixian Chen ◽  
Zhenhua Ji ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Squamous Cell Carcinoma ◽  
Cell Migration ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Cell Apoptosis ◽  
Intracellular Signaling ◽  
Migration And Invasion ◽  
Related Factor ◽  
Hypopharyngeal Squamous Cell Carcinoma

Background and Purpose: O-GlcNAcylation is a significant protein posttranslational modification with O-linked β-N-acetylglucosamine (GlcNAc) for intracellular signaling. Elevated O-GlcNAcylation contributes to cell proliferation, cell migration, cell apoptosis and signal transduction in various cancers. However, the expression level and functional role of O-GlcNAcylation in Hypopharyngeal Squamous Cell Carcinoma (HSCC) is not clearly elucidated. Nuclear factor erythroid-2-related factor 2 (Nrf2) is a master transcriptional factor that has been found to be aberrantly activated in HSCC. Here, we provide a molecular rationale between O-GlcNAcylation and Nrf2 in HSCC patients. Methods:: The protein levels of O-GlcNAcylation and Nrf2 in HSCC tissues were detected by immunohistochemistry technique and western blot analysis. Then, O‐GlcNAcylation knockdown HSCC cells were applied in this study. Cell proliferation was detected by CCK8, colony-forming analysis, and cell cycle assays. Cell migration and invasion ability was evaluated by transwell assays. Cell apoptosis was measured by TUNEL analysis. Results: O-GlcNAcylation was obviously up-regulated in HSCC tissues, which correlated with tumor size and lymph node metastasis. In addition, the protein level of Nrf2 was found to positively correlate with the expression of O‐GlcNAcylation both in vivo and in vitro. Knockdown of O-GlcNAcylation significantly inhibited HSCC cell growth, suppressed cell migration, and promoted cell apoptosis, whereas overexpression of Nrf2 reversed these phenotypes. Mechanismly, the upregulation of O-GlcNAcylation promoted the phosphorylation of Akt, leading to the stabilization of Nrf2; this could be attenuated by inhibition of the PI3K/Akt signaling pathway. Conclusion: Here, we provide a molecular association between O-GlcNAcylation and Nrf2 in HSCC patients, thus providing valuable therapeutic targets for the disease.

scholarly journals ARHGAP4-SEPT2-SEPT9 complex enables both up- and down-modulation of integrin-mediated focal adhesions, cell migration, and invasion

2021 ◽  
pp. mbc.E21-01-0010
Author(s):  
Kang Na ◽  
Tsubasa S. Matsui ◽  
Liu Shiyou ◽  
Shinji Deguchi
Keyword(s):  
Cell Migration ◽  
Rho Gtpase ◽  
Focal Adhesions ◽  
Migration And Invasion ◽  
Dependent Manner ◽  
Cell Migration And Invasion ◽  
Phenotypic Changes ◽  
Gtpase Activating Proteins ◽  
A Cell ◽  
Rho Family

The Rho family of GTPases are inactivated in a cell context-dependent manner by Rho-GTPase-activating proteins (Rho-GAPs), but their signaling mechanisms are poorly understood. Here we demonstrate that ARHGAP4, the Rho-GAPs, forms a complex with SEPT2 and SEPT9 via its Rho-GAP domain and SH3 domain to enable both up- and down-modulation of integrin-mediated focal adhesions (FAs). We show that silencing ARHGAP4 as well as overexpressing its two mutually independent upstream regulators SEPT2 and SEPT9 all induce reorganization of FAs to newly express Integrin Beta 1 and also enhance both cell migration and invasion. Interestingly, even if these cell migration/invasion-associated phenotypic changes are induced upon perturbations to the complex, it does not necessarily cause enhanced clustering of FAs. Instead, its extent depends on whether the microenvironment contains ligands suitable for the upregulated Integrin Beta 1. These results provide novel insights to cell migration, invasion, and microenvironment-dependent phenotypic changes regulated by the newly identified complex.

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