scholarly journals Modification of the Boyden Chamber to Improve Uniformity of Cell Invasion of Matrigel-Coated Membranes

BioTechniques ◽  
2001 ◽  
Vol 31 (6) ◽  
pp. 1242-1246 ◽  
Author(s):  
Kelly L. Bloomfield ◽  
Ben L. Baldwin ◽  
Damien G. Harkin ◽  
Kathryn F. Tonissen
Keyword(s):  
Cell Invasion ◽  
Boyden Chamber ◽  
Coated Membranes

scholarly journals MMP3 activity rather than cortical stiffness determines NHE1-dependent invasiveness of melanoma cells

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Dennis Keurhorst ◽  
Ivan Liashkovich ◽  
Fabian Frontzek ◽  
Svenja Nitzlaff ◽  
Verena Hofschröer ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Invasion ◽  
Time Lapse ◽  
Actin Dynamics ◽  
Cell Stiffness ◽  
Migration And Invasion ◽  
Boyden Chamber ◽  
Cortical Actin ◽  
Structural Element ◽  
Mmp Inhibitor

Abstract Background Both cell adhesion and matrix metalloproteinase (MMP) activity depend on pH at the cell surface. By regulating extracellular juxtamembrane pH, the Na+/H+ exchanger NHE1 plays a significant part in human melanoma (MV3) cell migration and invasion. Because NHE1, besides its pH-regulatory transport function, also serves as a structural element tying the cortical actin cytoskeleton to the plasma membrane, we investigated whether NHE1 affects cortical stiffness of MV3 cells, and how this makes an impact on their invasiveness. Methods NHE1 overexpressing MV3 cells were compared to the corresponding mock-transfected control cells. NHE1 expression was verified by Western blotting, cariporide (HOE642) was used to inhibit NHE1 activity, cell stiffness was determined by atomic force microscopy, and F-actin was visualized by phalloidin-staining. Migration on, and invasion of, native and glutaraldehyde-fixed collagen I substrates were analyzed using time-lapse video microscopy and Boyden-chamber assays, respectively. MMP secretion and activity were detected by Western blot and zymography, respectively. MMP activity was inhibited with NNGH. Results The cortical, but not the bulk stiffness, was significantly higher in NHE1 overexpressing cells. This increase in cortical stiffness was accompanied by a reorganization of the cortical cytoskeleton, i.e. a condensation of F-actin underneath and along the plasma membrane. However, it was not affected by NHE1 inhibition. Nevertheless, actin dynamics is required for cell invasion as demonstrated with the application of cytochalasin D. NHE1 overexpression was associated with an elevated MMP3 secretion and an increase in the invasion of a native matrix. This increase in invasiveness could be antagonized by the MMP inhibitor NNGH. Transmigration through a glutaraldehyde-fixed, indigestible substrate was not affected by NHE1 overexpression. Conclusion NHE1, as a structural element and independently of its transport activity, contributes to the organization of the cortical F-actin meshwork and thus impacts cortical stiffness. Since NHE1 overexpression stimulates MMP3 secretion but does not change transmigration through a fixed substrate, MV3 cell invasion of a native substrate depends on MMP activity rather than on a modifiable cortical stiffness.

scholarly journals Prodigiosin Inhibits Proliferation, Migration, and Invasion of Nasopharyngeal Cancer Cells

2018 ◽  
Vol 48 (4) ◽  
pp. 1556-1562 ◽  
Author(s):  
Yongze Liu ◽  
Han Zhou ◽  
Xiaofeng Ma ◽  
Chuanyao Lin ◽  
Ling Lu ◽  
...  
Keyword(s):  
Cell Migration ◽  
Nasopharyngeal Carcinoma ◽  
Tumor Cell ◽  
Cancer Cells ◽  
Cell Invasion ◽  
Nasopharyngeal Cancer ◽  
Migration And Invasion ◽  
Boyden Chamber ◽  
Cell Cycle Distribution ◽  
Tumor Cell Migration

Background/Aims: Nasopharyngeal carcinoma remains a devastating and difficult disease to treat. This study explores the antineoplastic effect of prodigiosin on nasopharyngeal cancer cells. Methods: Human nasopharyngeal carcinoma CNE2 cells and human normal nasopharyngeal epithelial NP69 cells were obtained and treated with prodigiosin or fluorouracil (5-FU). Colony formation assay was performed to screen for the optimal experimental concentrations of prodigiosin and 5-FU, and MTT assay was used to examine cell proliferative ability. Flow cytometry was used to examine cell cycle distribution, the scratch test was employed to examine cell migration, and Transwell migration assay (Boyden chamber) was used to study cell invasion. Results: The optimal concentrations of prodigiosin and 5-FU for treatment were 4 mg/L and 0.35 mg/L, respectively. Both prodigiosin and 5-FU inhibited tumor cell proliferation. The percentage of cells in G0/G1 phase was higher and the percentage of cells in S phase was lower in the prodigiosin and 5-FU groups than in the untreated groups. Both prodigiosin and 5-FU inhibited tumor cell migration and tumor cell invasion. Conclusions: Our results suggest that prodigiosin can inhibit proliferation, migration, and invasion of nasopharyngeal carcinoma cells.

scholarly journals Vital Stain to Study Cell Invasion in Modified Boyden Chamber Assay

BioTechniques ◽  
2002 ◽  
Vol 33 (6) ◽  
pp. 1200-1204 ◽  
Author(s):  
Madhura R. Vipra ◽  
Jayant M. Chiplonkar
Keyword(s):  
Cell Invasion ◽  
Boyden Chamber ◽  
Vital Stain ◽  
Boyden Chamber Assay

Kisspeptin-10 Inhibits Stromal-Derived Factor 1–Induced Invasion of Human Endometrial Cancer Cells

2014 ◽  
Vol 24 (2) ◽  
pp. 210-217 ◽  
Author(s):  
Elena Schmidt ◽  
Maike Haase ◽  
Elke Ziegler ◽  
Günter Emons ◽  
Carsten Gründker
Keyword(s):  
Endometrial Cancer ◽  
Basement Membrane ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Cell Invasion ◽  
Metastatic Cancer ◽  
Boyden Chamber ◽  
Cancer Cell Invasion ◽  
Endometrial Cancer Cell ◽  
Mg63 Cells

ObjectivesThe cross talk between metastatic cancer cells and target sites is critical for the development and progression of metastases. Disruption of this interaction will allow to design mechanism-based effective and specific therapeutic interventions for metastases. We have established a coculture system of cells derived from different tumor entities and MG63 human osteoblastlike cells to analyze tumor cell invasion. Recently, we have shown that breast cancer cell invasion was dramatically increased when cocultured with MG63 cells.Using this model, we have now analyzed whether stromal-derived factor 1 (SDF-1) is responsible for human endometrial cancer cell invasion and whether kisspeptin-10 (KP-10) treatment affects SDF-1–induced invasion of endometrial cancer cells in vitro.MethodsInvasion was quantified by assessment of endometrial cancer cell migration rate through an artificial basement membrane in a modified Boyden chamber during coculture with MG63 cells or after treatment with SDF-1α, SDF-1β, or the combination of both SDF-1 isoforms. In addition, the role of SDF-1 in invasion of endometrial cancer cells was analyzed by blocking SDF-1 secretion during coculture with MG64 cells. Furthermore, the effects of KP-10 treatment on MG63 coculture-driven and SDF-1–induced invasion were analyzed.ResultsEndometrial cancer cell invasion was significantly increased when cocultured with MG63 cells. Treatment with KP-10 reduced the ability to invade a reconstituted basement membrane and to migrate in response to the cellular stimulus. This effect was significant in a dose window of 10−13 to 10−11 mol/L. During coculture, SDF-1 protein expression of MG63 cells was significantly increased. The MG63 coculture-induced increase of endometrial cancer cell invasion could be blocked by anti–SDF-1 antibodies. Treatment of endometrial cancer cells in monoculture (without MG63) with SDF-1α, SDF-1β, or the combination of both isoforms resulted in a significant increase of endometrial cancer cell invasion. The SDF-1–induced increase of endometrial cancer cell invasion was significantly reduced after treatment with KP-10.ConclusionsOur findings suggest that SDF-1 plays a major role in endometrial cancer invasion. Stromal-derived factor 1–induced invasion can be inhibited by KP-10 treatment.

scholarly journals Tomatine Displays Antitumor Potential in In Vitro Models of Metastatic Melanoma

2020 ◽  
Vol 21 (15) ◽  
pp. 5243 ◽  
Author(s):  
Simona Serratì ◽  
Letizia Porcelli ◽  
Stefania Guida ◽  
Anna Ferretta ◽  
Rosa Maria Iacobazzi ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Cell Lines ◽  
Cell Invasion ◽  
Capillary Tube ◽  
Tube Formation ◽  
Boyden Chamber ◽  
Wild Type ◽  
Antitumor Potential ◽  
Vegf Release

There is a growing interest in the cytotoxic effects of bioactive glycoalkaloids, such as α-tomatine on tumor cells. Here, for the first time, we determine the antitumor potential of tomatine, a mixture of α-tomatine and dehydrotomatine, in metastatic melanoma (MM) cell lines harboring different BRAF and MC1R variants. We performed cytotoxicity experiments and annexin-V/propidium iodide staining to assess the apoptotic/necrotic status of the cells. ER stress and autophagy markers were revealed by Western Blot, whereas antiangiogenic and vascular-disrupting effects were evaluated through a capillary tube formation assay on matrigel and by ELISA kit for VEGF release determination. Cell invasion was determined by a Boyden chamber matrigel assay. Tomatine reduced 50% of cell viability and induced a concentration-dependent increase of apoptotic cells in the range of 0.5–1 μM in terms of α-tomatine. The extent of apoptosis was more than two-fold higher in V600BRAF-D184H/D184H MC1R cells than in BRAF wild-type cells and V600BRAF-MC1R wild-type cell lines. Additionally, tomatine increased the LC3I/II autophagy marker, p-eIF2α, and p-Erk1/2 levels in BRAF wild-type cells. Notably, tomatine strongly reduced cell invasion and melanoma-dependent angiogenesis by reducing VEGF release and tumor-stimulating effects on capillary tube formation. Collectively, our findings support tomatine as a potential antitumor agent in MM.

scholarly journals ESCIN MITIGATES HYPOXIA MIMICKING NCI-H23 CELLS THROUGH MODULATION OF MMPs, HIF-1α AND HIF-2α

2020 ◽  
pp. 26-30
Author(s):  
PANNEER SELVAM CHERMAKANI ◽  
GANAPASAM SUDHANDIRAN
Keyword(s):  
Cell Invasion ◽  
Therapeutic Agent ◽  
Gelatin Zymography ◽  
Immunoblot Analysis ◽  
Migration And Invasion ◽  
Boyden Chamber ◽  
Dependent Manner ◽  
Lung Cancer Cell ◽  
Chemically Induced ◽  
Dose Dependent

Objective: The objective of the study is to investigate the effect of escin in hypoxia mimicked NCI-H23 cells through the modulation of matrix metalloproteinases (MMPs) 2 and 9. Methods: In escin-treated NCI-H23 cells, adhesion, migration, and invasion were detected by the adhesion, wound healing, and Boyden chamber assays, respectively. The activation of proteinases was detected using zymography assay. The expressions of HIF-1α and HIF-2α were evaluated by immunoblot. Results: In the present study, it was observed that escin suppressed chemically induced hypoxia condition and stimulated adhesion, migration, and invasion of NCI-H23 cells. Gelatin zymography assay showed that escin inhibited CoCl2 induced MMPs-2 and 9 activations in NCI-H23 cells. Furthermore, immunoblot analysis revealed that escin treatment decreased the expression of both HIF-1 and 2α in a dose-dependent manner under CoCl2 induced hypoxia condition. Conclusion: Taken together, these results indicate that escin inhibits HIFs-α mediated MMPs-2 and 9 expressions, resulting in suppression of lung cancer cell invasion that is induced by chemically induced hypoxia condition. Escin is a potential therapeutic agent for clinical use in preventing the invasion of human malignant lung tumors.

The Inhibitory Effect of Heparin and Related Glycosaminoglycans on Neutrophil Chemotaxis

1984 ◽  
Vol 52 (02) ◽  
pp. 134-137 ◽  
Author(s):  
Yaacov Matzner ◽  
Gerard Marx ◽  
Ruth Drexler ◽  
Amiram Eldor
Keyword(s):  
Specific Binding ◽  
Anticoagulant Activity ◽  
Neutrophil Migration ◽  
Inhibitory Effect ◽  
Chemotactic Factor ◽  
Human Neutrophils ◽  
Boyden Chamber ◽  
Neutrophil Chemotaxis ◽  
Inhibitory Effects ◽  
Chemotactic Factors

SummaryClinical observations have shown that heparin has antiinflammatory activities. The effect of heparin on neutrophil chemotaxis was evaluated in vitro in the Boyden Chamber. This method enabled differentiation between the direct effects of heparin on neutrophil migration and locomotion, and its effects on chemotactic factors. Heparin inhibited both the random migration and directed locomotion of human neutrophils toward zymosan-activated serum (ZAS) and F-met-leu-phe (FMLP). Inhibition was found to be dependent on the concentrations of the heparin and of the chemotactic factors. No specific binding of heparin to the neutrophils could be demonstrated, and heparin’s inhibitory effects were eliminated by simple washing of the cells. When added directly to the chamber containing chemotactic factor, heparin inhibited the chemotactic activity of ZAS but not that of FMLP, suggesting a direct inhibitory effect against C5a, the principal chemotactic factor in ZAS.Experiments performed with low-molecular-weight heparin, N-desulfated heparin, dextran sulfate, chondroitin sulfate and dextran indicated that the inhibitory effects of heparin on neutrophil chemotaxis are not related to its anticoagulant activity, but probably depend on the degree of sulfation of the heparin molecule.

CUTL1–a modulator of tumor cell invasion and target of TGF-beta which is highly expressed in colon cancer

2004 ◽  
Vol 42 (08) ◽  
Author(s):  
P Michl ◽  
M Ei'Bahrawy ◽  
R Poulsom ◽  
A Ramjaun ◽  
J Downward
Keyword(s):  
Colon Cancer ◽  
Tumor Cell ◽  
Cell Invasion ◽  
Tumor Cell Invasion ◽  
Tgf Beta
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