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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 densities 250 × 103 cells/cm2 onto 1.5 and 4.0 mg/mL collagen I gels. ROCK-1 was knocked down via siRNA to study the impact of ROCK-I on cell invasion in a low- and high-density environment. ROCK-I knockdown resulted in a 60% increase in invasion for cells seeded in both 1.5 and 4.0 mg/mL scaffolds.
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fig6: SW620 cells were seeded at densities 250 × 103 cells/cm2 onto 1.5 and 4.0 mg/mL collagen I gels. ROCK-1 was knocked down via siRNA to study the impact of ROCK-I on cell invasion in a low- and high-density environment. ROCK-I knockdown resulted in a 60% increase in invasion for cells seeded in both 1.5 and 4.0 mg/mL scaffolds.

Mentions: SW620 cells were transfected with siRNA as described in Materials and Methods. Samples were tracked using a cell tracker dye. Increasing collagen concentration did seem to reduce overall invasion, though the effect was slight. Cells seeded onto 4.0 mg/mL collagen I gels had a ~20% less invasion depth than those seeded onto 1.5 mg/mL gels for both ROCK-I knockdown as well as untransfected cells. For both 1.5 and 4.0 mg/mL collagen I gels, ROCK-I knockdown resulted in a 1.6-fold increase in invasion depth compared to untransfected cells. Standard Student t-test was performed, and a P value of 0.05 was considered significant (∗). This data is shown in Figure 6.


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 densities 250 × 103 cells/cm2 onto 1.5 and 4.0 mg/mL collagen I gels. ROCK-1 was knocked down via siRNA to study the impact of ROCK-I on cell invasion in a low- and high-density environment. ROCK-I knockdown resulted in a 60% increase in invasion for cells seeded in both 1.5 and 4.0 mg/mL scaffolds.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368365&req=5

fig6: SW620 cells were seeded at densities 250 × 103 cells/cm2 onto 1.5 and 4.0 mg/mL collagen I gels. ROCK-1 was knocked down via siRNA to study the impact of ROCK-I on cell invasion in a low- and high-density environment. ROCK-I knockdown resulted in a 60% increase in invasion for cells seeded in both 1.5 and 4.0 mg/mL scaffolds.
Mentions: SW620 cells were transfected with siRNA as described in Materials and Methods. Samples were tracked using a cell tracker dye. Increasing collagen concentration did seem to reduce overall invasion, though the effect was slight. Cells seeded onto 4.0 mg/mL collagen I gels had a ~20% less invasion depth than those seeded onto 1.5 mg/mL gels for both ROCK-I knockdown as well as untransfected cells. For both 1.5 and 4.0 mg/mL collagen I gels, ROCK-I knockdown resulted in a 1.6-fold increase in invasion depth compared to untransfected cells. Standard Student t-test was performed, and a P value of 0.05 was considered significant (∗). This data is shown in Figure 6.

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