Phagocytized Beads Reduce the α5β1 Integrin Facilitated Invasiveness of Cancer Cells by Regulating Cellular Stiffness

Mierke, Claudia Tanja

doi:10.1007/s12013-012-9506-3

Cited by 17 publications

(14 citation statements)

References 70 publications

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“…An effect similar to interference with Rac signaling was also observed upon addition of a Rac1 inhibitor to MDA-MB-231 human breast cancer cells, in which the invasiveness of Rac1 inhibitor-treated cancer cells was pronouncedly reduced after three days 54 . Indeed, we could confirm these results, as we performed 3D motility assays in the presence and absence of the Rac1 inhibitor EHT1864 using Rac1 fl/fl fibroblast cells.…”

Section: Discussionmentioning

confidence: 68%

“…Cell mechanical properties such as stiffness and the generation of contractile forces have previously been implicated in driving cell motility and invasion in a one to two mixture of rat and bovine collagen fiber matrices 44,50,54–60 . Moreover, the deformability of cells measured with an optical cell stretcher correlates with their aggressiveness 47,61–63 .…”

Section: Resultsmentioning

confidence: 99%

“…After filling as yet unpolymerized collagen matrices into wells, gels were polymerized at 95% humidity under 5% CO 2 and at 37 °C, for approximately two hours. The fully polymerized gels were washed twice with PBS, and incubated overnight with DMEM complete growth medium 43,44,54 . In addition, we prepared 3 g/L collagen fiber matrices with the mixture of collagen R and collagen G in a 1 to 2 ratio, respectively.…”

Section: Methodsmentioning

confidence: 99%

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The Small GTPase Rac1 Increases Cell Surface Stiffness and Enhances 3D Migration Into Extracellular Matrices

Kunschmann

Puder

Fischer

et al. 2019

Sci Rep

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Membrane ruffling and lamellipodia formation promote the motility of adherent cells in two-dimensional motility assays by mechano-sensing of the microenvironment and initiation of focal adhesions towards their surroundings. Lamellipodium formation is stimulated by small Rho GTPases of the Rac subfamily, since genetic removal of these GTPases abolishes lamellipodium assembly. The relevance of lamellipodial or invadopodial structures for facilitating cellular mechanics and 3D cell motility is still unclear. Here, we hypothesized that Rac1 affects cell mechanics and facilitates 3D invasion. Thus, we explored whether fibroblasts that are genetically deficient for Rac1 (lacking Rac2 and Rac3) harbor altered mechanical properties, such as cellular deformability, intercellular adhesion forces and force exertion, and exhibit alterations in 3D motility. Rac1 knockout and control cells were analyzed for changes in deformability by applying an external force using an optical stretcher. Five Rac1 knockout cell lines were pronouncedly more deformable than Rac1 control cells upon stress application. Using AFM, we found that cell-cell adhesion forces are increased in Rac1 knockout compared to Rac1-expressing fibroblasts. Since mechanical deformability, cell-cell adhesion strength and 3D motility may be functionally connected, we investigated whether increased deformability of Rac1 knockout cells correlates with changes in 3D motility. All five Rac1 knockout clones displayed much lower 3D motility than Rac1-expressing controls. Moreover, force exertion was reduced in Rac1 knockout cells, as assessed by 3D fiber displacement analysis. Interference with cellular stiffness through blocking of actin polymerization by Latrunculin A could not further reduce invasion of Rac1 knockout cells. In contrast, Rac1-expressing controls treated with Latrunculin A were again more deformable and less invasive, suggesting actin polymerization is a major determinant of observed Rac1-dependent effects. Together, we propose that regulation of 3D motility by Rac1 partly involves cellular mechanics such as deformability and exertion of forces.

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Section: Discussionmentioning

confidence: 68%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The Small GTPase Rac1 Increases Cell Surface Stiffness and Enhances 3D Migration Into Extracellular Matrices

Kunschmann

Puder

Fischer

et al. 2019

Sci Rep

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“…After that time, beads were usually internalized by the cells. About 30 cells can be measured during the 40-min interval 57 .…”

Section: Methodsmentioning

confidence: 99%

Environmentally controlled magnetic nano-tweezer for living cells and extracellular matrices

Aermes

Hayn

Fischer

et al. 2020

Sci Rep

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The magnetic tweezer technique has become a versatile tool for unfolding or folding of individual molecules, mainly DNA. In addition to single molecule analysis, the magnetic tweezer can be used to analyze the mechanical properties of cells and extracellular matrices. We have established a magnetic tweezer that is capable of measuring the linear and non-linear viscoelastic behavior of a wide range of soft matter in precisely controlled environmental conditions, such as temperature, CO 2 and humidity. The magnetic tweezer presented in this study is suitable to detect specific differences in the mechanical properties of different cell lines, such as human breast cancer cells and mouse embryonic fibroblasts, as well as collagen matrices of distinct concentrations in the presence and absence of fibronectin crosslinks. The precise calibration and control mechanism employed in the presented magnetic tweezer setup provides the ability to apply physiological force up to 5 nN on 4.5 µm superparamagnetic beads coated with fibronectin and coupled to the cells or collagen matrices. These measurements reveal specific local linear and non-linear viscoelastic behavior of the investigated samples. The viscoelastic response of cells and collagen matrices to the force application is best described by a weak power law behavior. Our results demonstrate that the stress stiffening response and the fluidization of cells is cell type specific and varies largely between differently invasive and aggressive cancer cells. Finally, we showed that the viscoelastic behavior of collagen matrices with and without fibronectin crosslinks measured by the magnetic tweezer can be related to the microstructure of these matrices.

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“…Integrins are obligate heterodimers, which consist of αand β-subunits, in order to investigate whether Hsc70 affected integrin β1 specifically, the expression of the most common integrin α5 subunit was investigated. 14 As shown in Figure 2D and E, the results showed that both the mRNA level and protein expression on Hsc70-AS/U87 cell surface were not affected, suggesting the specificity of Hsc70 to integrin β1 subunit.…”

Section: Down-regulation Of Hsc70 In U87 Cells Promoted the Expressiomentioning

confidence: 80%

<p>Heat Shock Cognate Protein 70 Enhanced Integrin β1 Mediated Invasion in Cancer Cells</p>

Sun¹,

Cao²,

Guo³

et al. 2020

CMAR

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Glioblastoma is one of the most common malignant cancers worldwide. In our previous work, we have shown that heat shock cognate protein 70 (Hsc70) functions as a positive growth regulator in glioma. We investigated the role of Hsc70 in integrin β1 mediated invasion of glioma cells. Methods: In order to investigate whether the down-regulation of Hsc70 would affect the expression of integrin β1 subunit, HeLa cells were transiently transfected with Hsc70-AS or pcDNA3.0 vectors and the down-regulation of Hsc70 was confirmed by Western blotting. Human brain glioma U87 cells were stably transfected with Hsc70-AS or pcDNA3.0 vectors to further elucidate the relationship between Hsc70 and integrin β1 in human glioma cells. Cellular localization of integrin β1 was detected using immunofluorescence confocal microscopy analysis. Results: Here we reported that down-regulation of the expression of Hsc70 in U87 cells by transfection with antisense cDNA specifically increased the expression of cell surface integrin β1 without changing its mRNA. Meanwhile, the integrin β1 125-kD mature form increased while 105-kD precursor form decreased when Hsc70 was down-regulated. Mechanically, the U87 cells transfected with antisense cDNA of Hsc70 decreased the Golgi localization of integrin β1, strengthened its interaction with integrin α5 subunit, and enhanced the adhesion ability to fibronectin (FN) and the phosphorylation level of focal adhesion kinase (FAK). Conclusion: Overall, these results suggested that the down-regulation of Hsc70 expression could promote the expression of cell surface integrin β1 and subsequently inhibit glioma invasion phenotype.

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Phagocytized Beads Reduce the α5β1 Integrin Facilitated Invasiveness of Cancer Cells by Regulating Cellular Stiffness

Cited by 17 publications

References 70 publications

The Small GTPase Rac1 Increases Cell Surface Stiffness and Enhances 3D Migration Into Extracellular Matrices

The Small GTPase Rac1 Increases Cell Surface Stiffness and Enhances 3D Migration Into Extracellular Matrices

Environmentally controlled magnetic nano-tweezer for living cells and extracellular matrices

<p>Heat Shock Cognate Protein 70 Enhanced Integrin β1 Mediated Invasion in Cancer Cells</p>

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