Automated Multichamber Time-lapse Videography for Long-term In Vivo Observation of Migrating Cells

Bühler, H.; Abeln, T.; Díaz-Carballo, David; Nguemgo-Kouam, P.; Hero, Thomas; Adamietz, Irenäus A.

doi:10.21873/invivo.11063

Cited by 6 publications

(5 citation statements)

References 15 publications

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“…An established videography system [ 30 ] was used to investigate cell migration. The system allows observing cell migration over defined periods under normal cell-culture conditions (37°C, 5% CO 2 ).…”

Section: Methodsmentioning

confidence: 99%

Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

et al. 2018

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Glioblastoma is a primary brain tumor with a poor prognosis despite of many treatment regimens. Radiotherapy significantly prolongs patient survival and remains the most common treatment. Slit2 and Robo1 are evolutionarily conserved proteins involved in axon guidance, migration, and branching of neuronal cells. New studies have shown that Slit2 and Robo1 could play important roles in leukocyte chemotaxis and glioblastoma cell migration. Therefore, we investigated whether the Slit2/Robo1 complex has an impact on the motility of glioblastoma cells and whether irradiation with therapeutic doses modulates this effect. Our results indicate that photon irradiation increases the migration of glioblastoma cells in vitro. qPCR and immunoblotting experiments in two different glioblastoma cell lines (U-373 MG and U-87 MG) with different malignancy revealed that both Slit2 and Robo1 are significantly lower expressed in the cell populations with the highest motility and that the expression was reduced after irradiation. Overexpression of Robo1 significantly decreased the motility of glioblastoma cells and inhibited the accelerated migration of wild-type cells after irradiation. Immunoblotting analysis of migration-associated proteins (fascin and focal adhesion kinase) and of the epithelial-mesenchymal-transition-related protein vimentin showed that irradiation affected the migration of glioblastoma cells by increasing vimentin expression, which can be reversed by the overexpression of Slit2 and Robo1. Our findings suggest that Robo1 expression might counteract migration and also radiation-induced migration of glioblastoma cells, a process that might be connected to mesenchymal-epithelial transition.

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Section: Methodsmentioning

confidence: 99%

Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

et al. 2018

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“…Videography. Motility parameters were analysed with a selfconstructed videography system as described previously (16). For a videography sequence, the cells were seeded into 35mm Petri dishes, at a density of 16,000 cells per dish.…”

Section: Methodsmentioning

confidence: 99%

The Triple-negative Breast Cancer Cell Line MDA-MB 231 Is Specifically Inhibited by the Ionophore Salinomycin

et al. 2019

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Background/Aim: Tumour cells of the profile CD44 + /CD24 low/have a high tumorigenic potential. Salinomycin can specifically inhibit the growth of these cells. Herein, we investigated the effects of salinomycin on the viability and migration of triple negative breast cancer cells. Materials and Methods: We analysed two cell lines: i) triplenegative MDA-MB 231 breast cancer cells and ii) a cytokeratin 18-transfected, re-differentiated subclone of the MDA-MB 231 cell line. The viability was determined using an -2,5-diphenyltetrazolium bromide) test, and the migration was determined using 24-h videography. The expression of oestrogen receptor was determined using immunohistochemistry. Results: Salinomycin reduces all migration parameters in MDA-MB 231 cells. A significant correlation was found between increasing salinomycin concentrations and loss of cell viability, which was significantly less noticeable in the transfected control cells. Conclusion: With salinomycin there is a specific inhibition of MDA-MB 231 cells. Since MDA-MB 231 has over 90% cells with the profile CD44 + /CD24 low/-, these might represent a possible point of attack for salinomycin.

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“…The experiment was conducted under the following conditions: 6 mbar pressure, 2.5 ml/min flow rate and a shear stress of 1.9 dyn/cm 2 . With the help of our self-built video microscopy system [34], the circulating cells in the μ-slide Luer were observed. After completion of the experiment, all attached tumor cells were counted in three visual fields.…”

Section: Methodsmentioning

confidence: 99%

The increased adhesion of tumor cells to endothelial cells after irradiation can be reduced by FAK-inhibition

et al. 2019

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BackgroundRadiotherapy is administered in more than 60% of all solid tumors. Most patients are cured but a significant number develops local recurrences or distant metastases. The question arises if irradiation might influence the metastatic process. In the present study we examined whether the adhesion of glioblastoma or breast cancer cells to endothelial cells, an important step in metastasis, is affected by photon irradiation.MethodsU-87 MG, U-373 MG and MDA-MB-231 cancer cells as well as primary human endothelial cells were irradiated with 0, 2, 4, or 8 Gy photons at a dose rate of 5 Gy/min. The adhesion of cancer cells to endothelial cells was tested either with the Vybrant based assay via fluorescent labelling or with an ibidi pump system able to mimic the physiological blood flow in vitro. In addition, the impact of FAK (focal adhesion kinase) inhibitor PF-573, 228 on the adhesion of non-irradiated and irradiated tumor cells was analyzed. Adhesion related and regulated proteins were analyzed by Western blotting.ResultsThe cellular adhesion was increased after irradiation regardless of which cell type was irradiated. The FAK-inhibitor was able to reduce the adhesion of non-irradiated cells but also the irradiation-induced increase in adhesion of tumor cells to endothelium. Adhesion related proteins were enhanced after irradiation with 4 Gy or 8 Gy in both cells types. The increased adhesion after irradiation is accompanied by the phosphorylation of src (Y416), FAK (Y397) and increased expression of paxillin.ConclusionIrradiation with photons in therapeutic doses is able to enhance the interaction between tumor cells and endothelial cells and by that might influence important steps of the metastatic process.

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Automated Multichamber Time-lapse Videography for Long-term In Vivo Observation of Migrating Cells

Cited by 6 publications

References 15 publications

Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

The Triple-negative Breast Cancer Cell Line MDA-MB 231 Is Specifically Inhibited by the Ionophore Salinomycin

The increased adhesion of tumor cells to endothelial cells after irradiation can be reduced by FAK-inhibition

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