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Regulation of epithelial-mesenchymal transition in breast cancer cells by cell contact and adhesion.

Cichon MA, Nelson CM, Radisky DC - Cancer Inform (2015)

Bottom Line: We show here that EMT-related processes are linked to a broad and conserved program of transcriptional alterations that are influenced by cell contact and adhesion.We further find that treatment of cells with matrix metalloproteinase-3 (MMP-3), an inducer of EMT, interrupts a defined subset of cell contact-regulated genes, including genes encoding a variety of RNA splicing proteins known to regulate the expression of Rac1b, an activated splice isoform of Rac1 known to be a key mediator of MMP-3-induced EMT in breast, lung, and pancreas.These results provide new insights into how MMPs act in cancer progression and how loss of cell-cell interactions is a key step in the earliest stages of cancer development.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Mayo Clinic Cancer Center, Jacksonville, FL USA.

ABSTRACT
Epithelial-mesenchymal transition (EMT) is a physiological program that is activated during cancer cell invasion and metastasis. We show here that EMT-related processes are linked to a broad and conserved program of transcriptional alterations that are influenced by cell contact and adhesion. Using cultured human breast cancer and mouse mammary epithelial cells, we find that reduced cell density, conditions under which cell contact is reduced, leads to reduced expression of genes associated with mammary epithelial cell differentiation and increased expression of genes associated with breast cancer. We further find that treatment of cells with matrix metalloproteinase-3 (MMP-3), an inducer of EMT, interrupts a defined subset of cell contact-regulated genes, including genes encoding a variety of RNA splicing proteins known to regulate the expression of Rac1b, an activated splice isoform of Rac1 known to be a key mediator of MMP-3-induced EMT in breast, lung, and pancreas. These results provide new insights into how MMPs act in cancer progression and how loss of cell-cell interactions is a key step in the earliest stages of cancer development.

No MeSH data available.


Related in: MedlinePlus

Regulation of EMT characteristics by cell density in MCF10A cells. (A) Phase contrast micrographs of cells plated at indicated densities in 35-mm plates and imaged after 24 hours. Scale bar 200 μm. (B) Area of cells at indicated densities (n > 20 for each condition; values displayed as means ± SEM; ANOVA P < 0.001 for trend). (C–E) Quantitative PCR assessment of E-cadherin (C; ANOVA P = 0.042 for trend), N-cadherin (D; ANOVA P = 0.12 for trend), and vimentin (E; ANOVA P < 0.001 for trend) expression in the cell cultures.
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f1-cin-suppl.3-2015-001: Regulation of EMT characteristics by cell density in MCF10A cells. (A) Phase contrast micrographs of cells plated at indicated densities in 35-mm plates and imaged after 24 hours. Scale bar 200 μm. (B) Area of cells at indicated densities (n > 20 for each condition; values displayed as means ± SEM; ANOVA P < 0.001 for trend). (C–E) Quantitative PCR assessment of E-cadherin (C; ANOVA P = 0.042 for trend), N-cadherin (D; ANOVA P = 0.12 for trend), and vimentin (E; ANOVA P < 0.001 for trend) expression in the cell cultures.

Mentions: To determine how gradual differences in MCF10A cell density affected patterns of gene expression, cells were plated in 35-mm plates at 50K, 100K, 200K, 400K, 600K, and 800K cells per dish, and then cultured for 24 hours (Fig. 1A). Image analysis of cell morphology for these conditions (Fig. 1B) revealed that while the cells were more spread at the lowest densities, the differences between the higher densities were smaller. MCF10A cells are known to show differential expression of EMT marker genes depending on whether the cells are cultured under sparse or confluent conditions,13,40 and we also found differential expression of EMT markers, with progressively increasing mRNA levels of the epithelial marker E-cadherin (Fig. 1C) and progressively decreasing mRNA levels of the mesenchymal markers N-cadherin (Fig. 1D) and vimentin (Fig. 1E) at higher cell densities throughout the density range.


Regulation of epithelial-mesenchymal transition in breast cancer cells by cell contact and adhesion.

Cichon MA, Nelson CM, Radisky DC - Cancer Inform (2015)

Regulation of EMT characteristics by cell density in MCF10A cells. (A) Phase contrast micrographs of cells plated at indicated densities in 35-mm plates and imaged after 24 hours. Scale bar 200 μm. (B) Area of cells at indicated densities (n > 20 for each condition; values displayed as means ± SEM; ANOVA P < 0.001 for trend). (C–E) Quantitative PCR assessment of E-cadherin (C; ANOVA P = 0.042 for trend), N-cadherin (D; ANOVA P = 0.12 for trend), and vimentin (E; ANOVA P < 0.001 for trend) expression in the cell cultures.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4325704&req=5

f1-cin-suppl.3-2015-001: Regulation of EMT characteristics by cell density in MCF10A cells. (A) Phase contrast micrographs of cells plated at indicated densities in 35-mm plates and imaged after 24 hours. Scale bar 200 μm. (B) Area of cells at indicated densities (n > 20 for each condition; values displayed as means ± SEM; ANOVA P < 0.001 for trend). (C–E) Quantitative PCR assessment of E-cadherin (C; ANOVA P = 0.042 for trend), N-cadherin (D; ANOVA P = 0.12 for trend), and vimentin (E; ANOVA P < 0.001 for trend) expression in the cell cultures.
Mentions: To determine how gradual differences in MCF10A cell density affected patterns of gene expression, cells were plated in 35-mm plates at 50K, 100K, 200K, 400K, 600K, and 800K cells per dish, and then cultured for 24 hours (Fig. 1A). Image analysis of cell morphology for these conditions (Fig. 1B) revealed that while the cells were more spread at the lowest densities, the differences between the higher densities were smaller. MCF10A cells are known to show differential expression of EMT marker genes depending on whether the cells are cultured under sparse or confluent conditions,13,40 and we also found differential expression of EMT markers, with progressively increasing mRNA levels of the epithelial marker E-cadherin (Fig. 1C) and progressively decreasing mRNA levels of the mesenchymal markers N-cadherin (Fig. 1D) and vimentin (Fig. 1E) at higher cell densities throughout the density range.

Bottom Line: We show here that EMT-related processes are linked to a broad and conserved program of transcriptional alterations that are influenced by cell contact and adhesion.We further find that treatment of cells with matrix metalloproteinase-3 (MMP-3), an inducer of EMT, interrupts a defined subset of cell contact-regulated genes, including genes encoding a variety of RNA splicing proteins known to regulate the expression of Rac1b, an activated splice isoform of Rac1 known to be a key mediator of MMP-3-induced EMT in breast, lung, and pancreas.These results provide new insights into how MMPs act in cancer progression and how loss of cell-cell interactions is a key step in the earliest stages of cancer development.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Mayo Clinic Cancer Center, Jacksonville, FL USA.

ABSTRACT
Epithelial-mesenchymal transition (EMT) is a physiological program that is activated during cancer cell invasion and metastasis. We show here that EMT-related processes are linked to a broad and conserved program of transcriptional alterations that are influenced by cell contact and adhesion. Using cultured human breast cancer and mouse mammary epithelial cells, we find that reduced cell density, conditions under which cell contact is reduced, leads to reduced expression of genes associated with mammary epithelial cell differentiation and increased expression of genes associated with breast cancer. We further find that treatment of cells with matrix metalloproteinase-3 (MMP-3), an inducer of EMT, interrupts a defined subset of cell contact-regulated genes, including genes encoding a variety of RNA splicing proteins known to regulate the expression of Rac1b, an activated splice isoform of Rac1 known to be a key mediator of MMP-3-induced EMT in breast, lung, and pancreas. These results provide new insights into how MMPs act in cancer progression and how loss of cell-cell interactions is a key step in the earliest stages of cancer development.

No MeSH data available.


Related in: MedlinePlus