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Amino-terminal fragments of laminin γ2 chain retract vascular endothelial cells and increase vascular permeability.

Sato H, Oyanagi J, Komiya E, Ogawa T, Higashi S, Miyazaki K - Cancer Sci. (2014)

Bottom Line: Such effects of γ2pf led to prominent actin stress fiber formation in HUVECs, which was blocked by a ROCK inhibitor.As possible receptors, γ2pf interacted with heparan sulfate proteoglycans on the surface of HUVECs.Moreover, we localized the active site of γ2pf to the N-terminal epidermal growth factor-like repeat.

View Article: PubMed Central - PubMed

Affiliation: Department of Genome Science, Graduate School of Integrated Science and Nanobioscience, Yokohama City University, Yokohama, Japan; Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, Yokohama, Japan.

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Effect of laminin γ2 proteolytic fragment (γ2pf) on transendothelial migration of T-24 carcinoma cells. (a) Fluorescence-labeled Mock-T24 cells were incubated on a HUVEC monolayer with PBS (Control) or with γ2pf (2 μg/mL) in Transwell chambers. After incubation for 18 h, the cells that had migrated onto the lower surface of membrane filters were fixed and photographed under a fluorescence microscope. Small homogeneous spots are pores of the membrane filters. (b) Quantitative analysis of migrated cells. The areas of cells on the lower surface of membrane filters were measured by Image-J. Each point represents the mean ± SD (bar) of triplicate chambers. *P < 0.05. Essentially the same results were reproduced in an additional experiment.
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fig03: Effect of laminin γ2 proteolytic fragment (γ2pf) on transendothelial migration of T-24 carcinoma cells. (a) Fluorescence-labeled Mock-T24 cells were incubated on a HUVEC monolayer with PBS (Control) or with γ2pf (2 μg/mL) in Transwell chambers. After incubation for 18 h, the cells that had migrated onto the lower surface of membrane filters were fixed and photographed under a fluorescence microscope. Small homogeneous spots are pores of the membrane filters. (b) Quantitative analysis of migrated cells. The areas of cells on the lower surface of membrane filters were measured by Image-J. Each point represents the mean ± SD (bar) of triplicate chambers. *P < 0.05. Essentially the same results were reproduced in an additional experiment.

Mentions: The results shown above suggested that γ2pf might induce migration of cancer cells through the endothelial cell sheet. This was tested by the Transwell chamber assay. Fluorescence-labeled Mock-T24 cells were placed on the HUVEC monolayer in the upper chamber and incubated in the presence or absence of purified γ2pf. The number of cells that migrated through the endothelial monolayer to the lower chamber increased 2.5 times in the presence of γ2pf (Fig. 3a,b). This clearly indicated that γ2pf stimulated the cancer cell migration through the endothelial monolayer sheet.


Amino-terminal fragments of laminin γ2 chain retract vascular endothelial cells and increase vascular permeability.

Sato H, Oyanagi J, Komiya E, Ogawa T, Higashi S, Miyazaki K - Cancer Sci. (2014)

Effect of laminin γ2 proteolytic fragment (γ2pf) on transendothelial migration of T-24 carcinoma cells. (a) Fluorescence-labeled Mock-T24 cells were incubated on a HUVEC monolayer with PBS (Control) or with γ2pf (2 μg/mL) in Transwell chambers. After incubation for 18 h, the cells that had migrated onto the lower surface of membrane filters were fixed and photographed under a fluorescence microscope. Small homogeneous spots are pores of the membrane filters. (b) Quantitative analysis of migrated cells. The areas of cells on the lower surface of membrane filters were measured by Image-J. Each point represents the mean ± SD (bar) of triplicate chambers. *P < 0.05. Essentially the same results were reproduced in an additional experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317827&req=5

fig03: Effect of laminin γ2 proteolytic fragment (γ2pf) on transendothelial migration of T-24 carcinoma cells. (a) Fluorescence-labeled Mock-T24 cells were incubated on a HUVEC monolayer with PBS (Control) or with γ2pf (2 μg/mL) in Transwell chambers. After incubation for 18 h, the cells that had migrated onto the lower surface of membrane filters were fixed and photographed under a fluorescence microscope. Small homogeneous spots are pores of the membrane filters. (b) Quantitative analysis of migrated cells. The areas of cells on the lower surface of membrane filters were measured by Image-J. Each point represents the mean ± SD (bar) of triplicate chambers. *P < 0.05. Essentially the same results were reproduced in an additional experiment.
Mentions: The results shown above suggested that γ2pf might induce migration of cancer cells through the endothelial cell sheet. This was tested by the Transwell chamber assay. Fluorescence-labeled Mock-T24 cells were placed on the HUVEC monolayer in the upper chamber and incubated in the presence or absence of purified γ2pf. The number of cells that migrated through the endothelial monolayer to the lower chamber increased 2.5 times in the presence of γ2pf (Fig. 3a,b). This clearly indicated that γ2pf stimulated the cancer cell migration through the endothelial monolayer sheet.

Bottom Line: Such effects of γ2pf led to prominent actin stress fiber formation in HUVECs, which was blocked by a ROCK inhibitor.As possible receptors, γ2pf interacted with heparan sulfate proteoglycans on the surface of HUVECs.Moreover, we localized the active site of γ2pf to the N-terminal epidermal growth factor-like repeat.

View Article: PubMed Central - PubMed

Affiliation: Department of Genome Science, Graduate School of Integrated Science and Nanobioscience, Yokohama City University, Yokohama, Japan; Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, Yokohama, Japan.

Show MeSH
Related in: MedlinePlus