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Treatment with Y-27632, a ROCK Inhibitor, Increases the Proinvasive Nature of SW620 Cells on 3D Collagen Type 1 Matrix.

Vishnubhotla R, Bharadwaj S, Sun S, Metlushko V, Glover SC - Int J Cell Biol (2012)

Bottom Line: Increasing collagen I scaffold density resulted in a 2.5-fold increase in cell proliferation while treatment with Y-27632 attenuated this effect although 1.5 fold increase in cell invasion was observed in ROCK inhibited samples.Intriguingly, ROCK inhibition also resulted in a 3.5-fold increase in cell invasion within 3D collagen scaffolds for cells seeded at lower densities.This data suggests that although ROCK inhibitors have been used clinically to treat several medical conditions, its effect largely depends on the surrounding microenvironment.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Illinois at Chicago, Chicago, IL 60612, USA.

ABSTRACT
The concept of using tissue density as a mechanism to diagnose a tumor has been around for centuries. However, this concept has not been sufficiently explored in a laboratory setting. Therefore, in this paper, we observed the effects of cell density and extracellular matrix (ECM) density on colon cancer invasion and proliferation using SW620 cells. We also attempted to inhibit ROCK-I to determine its effect on cell invasion and proliferation using standard molecular biology techniques and advanced imaging. Increasing cell seeding density resulted in a 2-fold increase in cell invasion as well as cell proliferation independent of treatment with Y-27632. Increasing collagen I scaffold density resulted in a 2.5-fold increase in cell proliferation while treatment with Y-27632 attenuated this effect although 1.5 fold increase in cell invasion was observed in ROCK inhibited samples. Intriguingly, ROCK inhibition also resulted in a 3.5-fold increase in cell invasion within 3D collagen scaffolds for cells seeded at lower densities. We show in this paper that ROCK-I inhibition leads to increased invasion within 3D collagen I microenvironments. This data suggests that although ROCK inhibitors have been used clinically to treat several medical conditions, its effect largely depends on the surrounding microenvironment.

No MeSH data available.


Related in: MedlinePlus

SW620 cells were seeded at different densities: 50 × 103 (a), and 250 × 103 cells/cm2 (b). The lower seeding density (a) shows rounder cells with few cell-cell contacts. The greater seeding density allows for more cell-cell contact, creating a more epithelial state. Scale Bar = 10 μm.
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fig1: SW620 cells were seeded at different densities: 50 × 103 (a), and 250 × 103 cells/cm2 (b). The lower seeding density (a) shows rounder cells with few cell-cell contacts. The greater seeding density allows for more cell-cell contact, creating a more epithelial state. Scale Bar = 10 μm.

Mentions: Collagen scaffolds were prepared, and SW620 cells were seeded as described in the methods section. Cells were stained with phalloidin and imaged using a multiphoton microscope. Cells seeded at the lower cell density appeared to have relatively few cell-cell interactions. Within the collagen scaffold, these cells exhibited a symmetrically round phenotype. No protrusions or invadopodia were as observed in prior experiments using SW620 cells in 1.5 mg/mL scaffolds [20]. Cells seeded at the higher density, 4 mg/mL, were tightly packed within the 3D scaffold resulting in a forced cell-to-cell contact. It is important to note that the cells that appear “rounder” are on a plane above the rest of the cells. This can be seen in Figure 1.


Treatment with Y-27632, a ROCK Inhibitor, Increases the Proinvasive Nature of SW620 Cells on 3D Collagen Type 1 Matrix.

Vishnubhotla R, Bharadwaj S, Sun S, Metlushko V, Glover SC - Int J Cell Biol (2012)

SW620 cells were seeded at different densities: 50 × 103 (a), and 250 × 103 cells/cm2 (b). The lower seeding density (a) shows rounder cells with few cell-cell contacts. The greater seeding density allows for more cell-cell contact, creating a more epithelial state. Scale Bar = 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368365&req=5

fig1: SW620 cells were seeded at different densities: 50 × 103 (a), and 250 × 103 cells/cm2 (b). The lower seeding density (a) shows rounder cells with few cell-cell contacts. The greater seeding density allows for more cell-cell contact, creating a more epithelial state. Scale Bar = 10 μm.
Mentions: Collagen scaffolds were prepared, and SW620 cells were seeded as described in the methods section. Cells were stained with phalloidin and imaged using a multiphoton microscope. Cells seeded at the lower cell density appeared to have relatively few cell-cell interactions. Within the collagen scaffold, these cells exhibited a symmetrically round phenotype. No protrusions or invadopodia were as observed in prior experiments using SW620 cells in 1.5 mg/mL scaffolds [20]. Cells seeded at the higher density, 4 mg/mL, were tightly packed within the 3D scaffold resulting in a forced cell-to-cell contact. It is important to note that the cells that appear “rounder” are on a plane above the rest of the cells. This can be seen in Figure 1.

Bottom Line: Increasing collagen I scaffold density resulted in a 2.5-fold increase in cell proliferation while treatment with Y-27632 attenuated this effect although 1.5 fold increase in cell invasion was observed in ROCK inhibited samples.Intriguingly, ROCK inhibition also resulted in a 3.5-fold increase in cell invasion within 3D collagen scaffolds for cells seeded at lower densities.This data suggests that although ROCK inhibitors have been used clinically to treat several medical conditions, its effect largely depends on the surrounding microenvironment.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of Illinois at Chicago, Chicago, IL 60612, USA.

ABSTRACT
The concept of using tissue density as a mechanism to diagnose a tumor has been around for centuries. However, this concept has not been sufficiently explored in a laboratory setting. Therefore, in this paper, we observed the effects of cell density and extracellular matrix (ECM) density on colon cancer invasion and proliferation using SW620 cells. We also attempted to inhibit ROCK-I to determine its effect on cell invasion and proliferation using standard molecular biology techniques and advanced imaging. Increasing cell seeding density resulted in a 2-fold increase in cell invasion as well as cell proliferation independent of treatment with Y-27632. Increasing collagen I scaffold density resulted in a 2.5-fold increase in cell proliferation while treatment with Y-27632 attenuated this effect although 1.5 fold increase in cell invasion was observed in ROCK inhibited samples. Intriguingly, ROCK inhibition also resulted in a 3.5-fold increase in cell invasion within 3D collagen scaffolds for cells seeded at lower densities. We show in this paper that ROCK-I inhibition leads to increased invasion within 3D collagen I microenvironments. This data suggests that although ROCK inhibitors have been used clinically to treat several medical conditions, its effect largely depends on the surrounding microenvironment.

No MeSH data available.


Related in: MedlinePlus