Cooperative cell invasion: matrix metalloproteinase–mediated incorporation between cells

Mitchell, Camilla B.; O’Neill, Geraldine M.

doi:10.1091/mbc.e16-03-0194

Cited by 12 publications

(15 citation statements)

References 25 publications

(14 reference statements)

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“…Next, we analysed the effect of ATM‐3507 on spheroid growth and invasion. As previously established, the SKNSH spheroids displayed both multicellular strand invasion and elongated single cell invasion representing the mixed lineage of this cell line (Mitchell & O'Neill, ). Despite reduced detection of F‐actin, spheroid sizes were not significantly different in ATM‐3507 treated conditions (Figure a).…”

Section: Resultssupporting

confidence: 60%

“…Despite this, the different invasion phenotypes exhibited by SHEP, SKNSH and BE2C spheroids in 3D cultures may be relevant. SHEP spheroids display Rac‐dependent single cell invasion (Mitchell & O'Neill, ), SKNSH have a mix of single cell and collective strand invasion (Mitchell & O'Neill, ) and BE2C exhibit collective sheet invasion (this study). Therefore different responses to drug treatments may reflect differences between collective and single cell invasion.…”

Section: Discussionmentioning

confidence: 79%

“…Tropomyosin Tpm3.1 was detected using a sheep polyclonal anti‐γ9d antibody kindly provided by Prof Peter Gunning. Immunofluorescence of embedded cells and spheroids was performed as previously described (Mitchell & O'Neill, ). Confocal imaging of fixed and immunostained cells and spheroids was performed on a Leica SP5 II confocal microscope with a 10 x air objective.…”

Section: Methodsmentioning

confidence: 99%

“…Reflecting their cellular derivation, patient‐derived neuroblastomas exhibit collective invasion into local tissue in orthotopic mouse xenografts (Braekeveldt et al, ). Moreover, multicellular neuroblastoma spheroids embedded in 3D collagen display a range of cooperative and collective invasion behaviours (Mitchell & O'Neill, , ).…”

Section: Introductionmentioning

confidence: 99%

“…Collectively invading cells variously display sheets, strands, or small clumps of cells invading en masse (Friedl et al, 2012). In addition to the influence of direct cell-cell adhesion, cells can cooperatively invade with one population influencing the invasion of a second population of cells in a heterogeneous mix of cells (for example [Mitchell & O'Neill, 2016]). Tpm3.1 function in single cell migration and invasion has previously been demonstrated in B35 neuroblastoma cells (Bach et al, 2009;Bryce et al, 2003;Lees, Bach, Bradbury, et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

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Small molecule targeting of the actin associating protein tropomyosin Tpm3.1 increases neuroblastoma cell response to inhibition of Rac‐mediated multicellular invasion

2018

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The migration and invasion of cells through tissues in the body is facilitated by a dynamic actin cytoskeleton. The actin-associating protein, tropomyosin Tpm3.1 has emerged to play important roles in cell migration and invasion. To date, investigations have focused on single cell migration and invasion where Tpm3.1 expression is inversely associated with Rac GTPase-mediated cell invasion. While single cell and collective cell invasion have many features in common, collective invasion is additionally impacted by cell-cell adhesion, and the role of Tpm3.1 in collective invasion has not been established. In the present study we have modelled multicellular invasion using neuroblastoma spheroids embedded in 3D collagen and analysed the function of Tpm3.1 using recently established compounds that target the Tpm3.1 C-terminus. The major findings from our study reveal that combined Rac inhibition and Tpm3.1 targeting result in greater inhibition of multicellular invasion than either treatment alone. Together, the data suggest that Tpm3.1 disruption sensitises neuroblastoma cells to inhibition of Rac-mediated multicellular invasion.

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Section: Resultssupporting

confidence: 60%

Section: Discussionmentioning

confidence: 79%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Small molecule targeting of the actin associating protein tropomyosin Tpm3.1 increases neuroblastoma cell response to inhibition of Rac‐mediated multicellular invasion

2018

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Annexin A1 peptide and endothelial cell‐conditioned medium modulate cervical tumorigenesis

et al. 2019

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Cervical cancer is one of the leading causes of cancer death in women worldwide, and its tumorigenesis can be influenced by the microenvironment. The anti‐inflammatory protein annexin A1 (ANXA1) has been reported to be associated with cancer progression and metastasis, suggesting that it plays a role in regulating tumour cell proliferation. Here, we examined the effect of the N‐terminal peptide Ac2‐26 of ANXA1 on the HaCaT cell line (normal) and HeLa cell line (cervical cancer) co‐cultured with endothelium cell‐conditioned medium (HMC). Treatment with Ac2‐26 decreased proliferation and increased motility of cervical cancer cells, but did not affect cellular morphology or viability. Combined HMC stimulus and Ac2‐26 treatment resulted in an increase in apoptotic HeLa cells, upregulated expression of MMP2, and downregulated expression of COX2, EP3 and EP4. In conclusion, Ac2‐26 treatment may modulate cellular and molecular mechanisms underlying cervical carcinogenesis.

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Analysis of Invasion Dynamics of Matrix-Embedded Cells in a Multisample Format

Troys¹,

Masuzzo²,

Huyck³

et al. 2018

Methods in Molecular Biology

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In vitro tests of cancer cell invasion are the "first line" tools of preclinical researchers for screening the multitude of chemical compounds or cell perturbations that may aid in halting or treating cancer malignancy. In order to have predictive value or to contribute to designing personalized treatment regimes, these tests need to take into account the cancer cell environment and measure effects on invasion in sufficient detail. The in vitro invasion assays presented here are a trade-off between feasibility in a multisample format and mimicking the complexity of the tumor microenvironment. They allow testing multiple samples and conditions in parallel using 3D-matrix-embedded cells and deal with the heterogeneous behavior of an invading cell population in time. We describe the steps to take, the technical problems to tackle and useful software tools for the entire workflow: from the experimental setup to the quantification of the invasive capacity of the cells. The protocol is intended to guide researchers to standardize experimental set-ups and to annotate their invasion experiments in sufficient detail. In addition, it provides options for image processing and a solution for storage, visualization, quantitative analysis, and multisample comparison of acquired cell invasion data.

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Cooperative cell invasion: matrix metalloproteinase–mediated incorporation between cells

Abstract: Cooperative invasion is observed in heterogeneous mixes of cancer cells. Two distinct tumor cell subtypes, neuroblastoma neuronal- and substrate-type cells, are shown to mutually influence each other’s invasion.

Cited by 12 publications

References 25 publications

Small molecule targeting of the actin associating protein tropomyosin Tpm3.1 increases neuroblastoma cell response to inhibition of Rac‐mediated multicellular invasion

Small molecule targeting of the actin associating protein tropomyosin Tpm3.1 increases neuroblastoma cell response to inhibition of Rac‐mediated multicellular invasion

Annexin A1 peptide and endothelial cell‐conditioned medium modulate cervical tumorigenesis

Analysis of Invasion Dynamics of Matrix-Embedded Cells in a Multisample Format

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