Thymosin beta4 inhibits ADF/cofilin stimulated F-actin cycling and hela cell migration: Reversal by active Arp2/3 complex

During their passage through the circulatory system, tumor cells undergo extensive interactions with various host cells including endothelial cells and platelets. Mechanisms mediating tumor cell adhesion, migration, and metastasis to vessel wall under flow condition are largely unknown. The aim of this study was to investigate the potential roles of GPIIb/IIIa andαvβ3integrins underlying the HeLa-endothelium interaction in static and dynamic flow conditions. HeLa cell migration and invasion were studied by using Millicell cell culture insert system. The numbers of transmigrated or invaded HeLa cells significantly increased by thrombin-activated platelets and reduced by eptifibatide, a platelet inhibitor. Meanwhile, RGDWE peptides, a specific inhibitor ofαvβ3integrin, also inhibited HeLa cell transmigration. Interestingly, the presence of endothelial cells had significant effect on HeLa cell migration regardless of static or cocultured flow condition. The adhesion capability of HeLa cells to endothelial monolayer was also significantly affected by GPIIb/IIIa andαvβ3integrins. The arrested HeLa cells increased nearly 5-fold in the presence of thrombin-activated platelets at shear stress condition (1.84 dyn/cm2exposure for 1 hour) than the control (static). Our findings showed that GPIIb/IIIa andαvβ3integrins are important mediators in the pathology of cervical cancer and provide a molecular basis for the future therapy, and the efficient antitumor benefit should target multiple receptors on tumor cells and platelets.

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TTBK2 with EB1/3 regulates microtubule dynamics in migrating cells through KIF2A phosphorylation

The Journal of Cell Biology ◽

10.1083/jcb.201412075 ◽

2015 ◽

Vol 210 (5) ◽

pp. 737-751 ◽

Cited By ~ 26

Author(s):

Takashi Watanabe ◽

Mai Kakeno ◽

Toshinori Matsui ◽

Ikuko Sugiyama ◽

Nariko Arimura ◽

...

Keyword(s):

Cell Migration ◽

Hela Cell ◽

Binding Proteins ◽

Microtubule Dynamics ◽

Wild Type ◽

Intact Cells ◽

Cellular Events ◽

Migrating Cells

Microtubules (MTs) play critical roles in various cellular events, including cell migration. End-binding proteins (EBs) accumulate at the ends of growing MTs and regulate MT end dynamics by recruiting other plus end–tracking proteins (+TIPs). However, how EBs contribute to MT dynamics through +TIPs remains elusive. We focused on tau-tubulin kinase 2 (TTBK2) as an EB1/3-binding kinase and confirmed that TTBK2 acted as a +TIP. We identified MT-depolymerizing kinesin KIF2A as a novel substrate of TTBK2. TTBK2 phosphorylated KIF2A at S135 in intact cells in an EB1/3-dependent fashion and inactivated its MT-depolymerizing activity in vitro. TTBK2 depletion reduced MT lifetime (facilitated shrinkage and suppressed rescue) and impaired HeLa cell migration, and these phenotypes were partially restored by KIF2A co-depletion. Expression of nonphosphorylatable KIF2A, but not wild-type KIF2A, reduced MT lifetime and slowed down the cell migration. These findings indicate that TTBK2 with EB1/3 phosphorylates KIF2A and antagonizes KIF2A-induced depolymerization at MT plus ends for cell migration.

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Volume-activated chloride channels contribute to cell-cycle-dependent regulation of HeLa cell migration

Biochemical Pharmacology ◽

10.1016/j.bcp.2008.10.009 ◽

2009 ◽

Vol 77 (2) ◽

pp. 159-168 ◽

Cited By ~ 34

Author(s):

Jianwen Mao ◽

Lixin Chen ◽

Bin Xu ◽

Lijing Wang ◽

Weizhang Wang ◽

...

Keyword(s):

Cell Cycle ◽

Cell Migration ◽

Hela Cell ◽

Chloride Channels ◽

Cycle Dependent

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Analysis of vascular smooth muscle cell and HeLa cell migration on the microgrooved substrate (Cell type differences of the mechanical sensing for microgrooved surfaces)

Transactions of the JSME (in Japanese) ◽

10.1299/transjsme.20-00301 ◽

2020 ◽

Vol 86 (892) ◽

pp. 20-00301-20-00301

Author(s):

Tatsuya HANZAWA ◽

Kazuaki NAGAYAMA

Keyword(s):

Smooth Muscle ◽

Cell Migration ◽

Vascular Smooth Muscle ◽

Hela Cell ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Cell Type ◽

Mechanical Sensing

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HGAL - A Germinal Center-Like Diffuse Large B-Cell Lymphoma Prognostic Biomarker Interacts with the Cytoskeleton and Influences Cell Migration.

Blood ◽

10.1182/blood.v108.11.2022.2022 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2022-2022

Author(s):

Jun Chen ◽

Feiying Ding ◽

David M. Helfman ◽

Izidore S. Lossos

Keyword(s):

Cell Migration ◽

Hela Cell ◽

Tyrosine Phosphorylation ◽

Cell Lines ◽

Hela Cells ◽

Germinal Center ◽

B Cell Lymphoma ◽

Lymphocyte Migration ◽

Transfected Hela Cell ◽

Hela Cell Lines

Abstract Gene expression profiling studies of lymphoid malignancies have led to the discovery of previously unrecognized lymphoma subtypes and associated novel genes. We recently cloned a germinal center (GC) specific gene - human germinal center-associated lymphoma (HGAL), whose expression correlates with survival in patients with diffuse large B-cell lymphoma (DLBCL) and classical Hodgkin lymphoma (HL). HGAL protein contains an ITAM motif and harbors 6 tyrosines that may be potentially phosphorylated. Mice deficient in the HGAL homologue M17 form normal GCs with the exception of reduced-sized Peyer’s patches, undergo efficient class-switch recombination and somatic hypermutation, and mount T-cell-dependent antibody responses similar to wild-type controls (Schenten et al., Blood 2006). The biological functions of HGAL in normal GC lymphocytes and lymphoma are unknown. Herein we demonstrate that HGAL plays a role in regulation of cell migration. Ex-vivo exposure of lymphoma or HGAL-transfected HeLa cell-lines to pervanadate resulted in tyrosine phosphorylation of HGAL protein. Stimulation of lymphoma and HGAL-transfected HeLa cell-lines with interleukin (IL)-6 led to transient HGAL tyrosine phosphorylation, peaking at 5–15 minutes. The phosphorylated HGAL had a markedly shortened life time compared to the unphosphorylated protein. No HGAL tyrosine phosphorylation was observed upon stimulation with anti IgM, interferon-g or IL-4. Deletion of the two C-terminal ITAM tyrosines in a truncated HGAL mutant (amino-acids 1-118) totally abrogated HGAL tyrosine phosphorylation upon IL-6 stimulation or exposure to pervanadate. Immunofluorescence and confocal miscroscopic studies of the SUDHL6 lymphoma and transfected HeLa cell-lines demonstrated HGAL redistribution from mainly perinuclear cytoplasmic localization to podosomes and spike-like filopodia. The time-frame of the HGAL relocalization was similar to that of HGAL tyrosine phosphorylation, being most prominent at 5 and 10 minutes in the SUDHL6 and transfected HeLa cells, respectively. Immunofluorescence studies demonstrated HGAL colocalization with actin, myosin and WASP proteins. HGAL immunoprecipitation studies followed by MAS spectroscopy or Western blotting confirmed HGAL interaction with actin and myosin type 2, but not with WASP protein. The N terminal portion of HGAL protein (aa 1-118) was sufficient for these interactions, however HGAL phosphorylation at its C-terminal tyrosines increased and extended the duration of its interaction with the myosin type 2. We further assessed the effects of HGAL on cell migration. Upon IL-6 stimulation for 24 hours, the migration of the Neo-HeLa cells in wound assay was markedly increased. However this effect of IL-6 was significantly ameliorated in the HGAL-HeLa cells. Moreover, siRNA knockdown of the HGAL in the HGAL-HeLa cells markedly reversed the HGAL-induced inhibition of IL-6 stimulated cell migration. This IL-6 effect on HGAL-HeLa and control Neo-HeLa cells was not attributable to differences in cell proliferation. Collectively, our results suggest that HGAL is involved in negative regulation of lymphocyte migration, thus constraining lymphocytes to the GC. Furthermore, inhibition of lymphocyte migration might contribute to the less aggressive clinical behavior of HGAL expressing DLBCL and HL tumors.

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