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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

Analysis of genes differentially expressed by density in MCF10A cells. (A–D) Genes upregulated more than two-fold in cells cultured at 800K density vs 50K density (A and B; n = 1444 features mapped to 1131 genes) or downregulated more than two-fold in cells cultured at 800K density vs 50K density (C and D; n = 1658 features mapped to 1303 genes); all genes are normalized to 50K expression and displayed as line graphs (A and C; colored by expression at 800K) or box-and-whisker plots (B and D). (E and F) Overlap of dataset of genes differentially regulated two-fold in MCF10A cells cultured at 800K density vs 50K density with datasets of genes differentially regulated between MDA-MB-231 cells and MCF10A cells (showing negative correlation; E) and of genes differentially regulated between MCF10A cells cultured on differentiating conditions vs 2D monolayers (showing positive correlation; F).
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f2-cin-suppl.3-2015-001: Analysis of genes differentially expressed by density in MCF10A cells. (A–D) Genes upregulated more than two-fold in cells cultured at 800K density vs 50K density (A and B; n = 1444 features mapped to 1131 genes) or downregulated more than two-fold in cells cultured at 800K density vs 50K density (C and D; n = 1658 features mapped to 1303 genes); all genes are normalized to 50K expression and displayed as line graphs (A and C; colored by expression at 800K) or box-and-whisker plots (B and D). (E and F) Overlap of dataset of genes differentially regulated two-fold in MCF10A cells cultured at 800K density vs 50K density with datasets of genes differentially regulated between MDA-MB-231 cells and MCF10A cells (showing negative correlation; E) and of genes differentially regulated between MCF10A cells cultured on differentiating conditions vs 2D monolayers (showing positive correlation; F).

Mentions: Transcriptional profiling of the MCF10A cells identified a large number of genes that were substantially regulated by cell density: 1444 probes (mapped to 1131 genes) were upregulated more than two-fold in 800K density relative to 50K density (Fig. 2A and B) and 1658 probes (mapped to 1303 genes) were downregulated more than two-fold in 800K density relative to 50K density (Fig. 2C and D). Meta-analysis of the MCF10A gene set using the NextBio platform revealed significant overlap with numerous cancer-associated datasets (Table 1), including lung cancer, liver cancer, and breast cancer. The negative association of the MCF10A dataset (comparing 800K density vs 50K density) with most cancer datasets indicates that lower cell density is more associated with cancer as compared with the normal tissue. Examination of overlap with individual datasets (Table 2) revealed significant negative correlation between the MCF10A dataset and gene sets comparing breast cancer vs normal breast tissue,41–46 particularly when the breast cancers were of the basal molecular subtype. This finding suggests that the MCF10A cells, which are immortal and nontransformed but which are classified as basal-type breast cancer cells,13,47 activate basal-type cancer characteristics at lower cell densities. The MCF10A dataset also showed significant negative overlap with comparisons of breast cancer cell lines, including MCF10A cells expressing activated ErbB2 vs control vector,48 the transformed MDA-MB-231, MCF-7 and T47D breast cancer cell lines vs MCF10A cells,49 and the MDA-MB-231 cells vs M98040 normal breast cells.50 With regard to the highly significant overlap (P = 1.9E − 184) with the dataset comparing MDA-MB-231 cells vs MCF10A cells, it was striking that the majority of the 2385 overlapping gene features showed a negative correlation (Fig. 2E), indicating that more than 2/3 of the transcriptional alterations induced by low density cultivation of MCF10A cells overlapped significantly with MDA-MB-231 cells. Comparison of the dataset of density-dependent gene expression changes in MCF10A cells with clinical datasets examining differences between breast cancers of different intrinsic subtypes identified signifi-cant overlap with four datasets comparing basal subtype breast cancers with normal breast tissue (Supplementary Fig. 1A, P = 1.6E − 65, ref.42; Supplementary Fig. 1B, P = 15.0E − 63, ref. 51; Supplementary Fig. 1C, P = 1.2E − 28, ref.52; and Supplementary Fig. 1D, P = 1.6E − 48, ref.53). We also identified significant overlap with a dataset comparing basal subtype breast cancer vs normal subtype breast cancer (Supplementary Fig. 1E, P = 2.2E − 25, ref.52), and another dataset comparing breast cancer cell lines of the basal B subtype vs basal A subtype (Supplementary Fig. 1F, P = 3.7E − 54, ref.54). The direction of association indicates that MCF10A cells plated at low density manifest an increasing association with basal subtype breast cancer.


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

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

Analysis of genes differentially expressed by density in MCF10A cells. (A–D) Genes upregulated more than two-fold in cells cultured at 800K density vs 50K density (A and B; n = 1444 features mapped to 1131 genes) or downregulated more than two-fold in cells cultured at 800K density vs 50K density (C and D; n = 1658 features mapped to 1303 genes); all genes are normalized to 50K expression and displayed as line graphs (A and C; colored by expression at 800K) or box-and-whisker plots (B and D). (E and F) Overlap of dataset of genes differentially regulated two-fold in MCF10A cells cultured at 800K density vs 50K density with datasets of genes differentially regulated between MDA-MB-231 cells and MCF10A cells (showing negative correlation; E) and of genes differentially regulated between MCF10A cells cultured on differentiating conditions vs 2D monolayers (showing positive correlation; F).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4325704&req=5

f2-cin-suppl.3-2015-001: Analysis of genes differentially expressed by density in MCF10A cells. (A–D) Genes upregulated more than two-fold in cells cultured at 800K density vs 50K density (A and B; n = 1444 features mapped to 1131 genes) or downregulated more than two-fold in cells cultured at 800K density vs 50K density (C and D; n = 1658 features mapped to 1303 genes); all genes are normalized to 50K expression and displayed as line graphs (A and C; colored by expression at 800K) or box-and-whisker plots (B and D). (E and F) Overlap of dataset of genes differentially regulated two-fold in MCF10A cells cultured at 800K density vs 50K density with datasets of genes differentially regulated between MDA-MB-231 cells and MCF10A cells (showing negative correlation; E) and of genes differentially regulated between MCF10A cells cultured on differentiating conditions vs 2D monolayers (showing positive correlation; F).
Mentions: Transcriptional profiling of the MCF10A cells identified a large number of genes that were substantially regulated by cell density: 1444 probes (mapped to 1131 genes) were upregulated more than two-fold in 800K density relative to 50K density (Fig. 2A and B) and 1658 probes (mapped to 1303 genes) were downregulated more than two-fold in 800K density relative to 50K density (Fig. 2C and D). Meta-analysis of the MCF10A gene set using the NextBio platform revealed significant overlap with numerous cancer-associated datasets (Table 1), including lung cancer, liver cancer, and breast cancer. The negative association of the MCF10A dataset (comparing 800K density vs 50K density) with most cancer datasets indicates that lower cell density is more associated with cancer as compared with the normal tissue. Examination of overlap with individual datasets (Table 2) revealed significant negative correlation between the MCF10A dataset and gene sets comparing breast cancer vs normal breast tissue,41–46 particularly when the breast cancers were of the basal molecular subtype. This finding suggests that the MCF10A cells, which are immortal and nontransformed but which are classified as basal-type breast cancer cells,13,47 activate basal-type cancer characteristics at lower cell densities. The MCF10A dataset also showed significant negative overlap with comparisons of breast cancer cell lines, including MCF10A cells expressing activated ErbB2 vs control vector,48 the transformed MDA-MB-231, MCF-7 and T47D breast cancer cell lines vs MCF10A cells,49 and the MDA-MB-231 cells vs M98040 normal breast cells.50 With regard to the highly significant overlap (P = 1.9E − 184) with the dataset comparing MDA-MB-231 cells vs MCF10A cells, it was striking that the majority of the 2385 overlapping gene features showed a negative correlation (Fig. 2E), indicating that more than 2/3 of the transcriptional alterations induced by low density cultivation of MCF10A cells overlapped significantly with MDA-MB-231 cells. Comparison of the dataset of density-dependent gene expression changes in MCF10A cells with clinical datasets examining differences between breast cancers of different intrinsic subtypes identified signifi-cant overlap with four datasets comparing basal subtype breast cancers with normal breast tissue (Supplementary Fig. 1A, P = 1.6E − 65, ref.42; Supplementary Fig. 1B, P = 15.0E − 63, ref. 51; Supplementary Fig. 1C, P = 1.2E − 28, ref.52; and Supplementary Fig. 1D, P = 1.6E − 48, ref.53). We also identified significant overlap with a dataset comparing basal subtype breast cancer vs normal subtype breast cancer (Supplementary Fig. 1E, P = 2.2E − 25, ref.52), and another dataset comparing breast cancer cell lines of the basal B subtype vs basal A subtype (Supplementary Fig. 1F, P = 3.7E − 54, ref.54). The direction of association indicates that MCF10A cells plated at low density manifest an increasing association with basal subtype breast cancer.

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