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Inflammation Mediated Metastasis: Immune Induced Epithelial-To-Mesenchymal Transition in Inflammatory Breast Cancer Cells.

Cohen EN, Gao H, Anfossi S, Mego M, Reddy NG, Debeb B, Giordano A, Tin S, Wu Q, Garza RJ, Cristofanilli M, Mani SA, Croix DA, Ueno NT, Woodward WA, Luthra R, Krishnamurthy S, Reuben JM - PLoS ONE (2015)

Bottom Line: It is unknown whether immune cells associated to the IBC microenvironment play a role in this scenario to transiently promote epithelial to mesenchymal transition (EMT) in these cells.Interestingly, although IBC cells exhibited increased invasion and migration following exposure to immune factors, the expression of E-cadherin (CDH1), a cell adhesion molecule, increased uniquely in IBC cell lines but not in non-IBC cell lines.These data suggest that release of cytokines by activated immune cells may contribute to the aggressiveness of IBC and highlight these factors as potential target mediators of immune-IBC interaction.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; The Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; The University of Texas Graduate School of Biomedical Sciences at Houston, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.

ABSTRACT
Inflammatory breast cancer (IBC) is the most insidious form of locally advanced breast cancer; about a third of patients have distant metastasis at initial staging. Emerging evidence suggests that host factors in the tumor microenvironment may interact with underlying IBC cells to make them aggressive. It is unknown whether immune cells associated to the IBC microenvironment play a role in this scenario to transiently promote epithelial to mesenchymal transition (EMT) in these cells. We hypothesized that soluble factors secreted by activated immune cells can induce an EMT in IBC and thus promote metastasis. In a pilot study of 16 breast cancer patients, TNF-α production by peripheral blood T cells was correlated with the detection of circulating tumor cells expressing EMT markers. In a variety of IBC model cell lines, soluble factors from activated T cells induced expression of EMT-related genes, including FN1, VIM, TGM2, ZEB1. Interestingly, although IBC cells exhibited increased invasion and migration following exposure to immune factors, the expression of E-cadherin (CDH1), a cell adhesion molecule, increased uniquely in IBC cell lines but not in non-IBC cell lines. A combination of TNF-α, IL-6, and TGF-β was able to recapitulate EMT induction in IBC, and conditioned media preloaded with neutralizing antibodies against these factors exhibited decreased EMT. These data suggest that release of cytokines by activated immune cells may contribute to the aggressiveness of IBC and highlight these factors as potential target mediators of immune-IBC interaction.

No MeSH data available.


Related in: MedlinePlus

Conditioned media from activated healthy donor PBMC induces EMT in IBC.(a) Morphological changes visualized by bright field in IBC cell lines consistent with stress and EMT were observed following 48-hour incubation with CM. (b) Immunohistochemical stains of paraffin cell blocks: the percentage of positive cells is listed in each image and shown in the bar graph at right. Pancytokeratin expression is shown for both percent of cells with membrane localization (top number) and cytoplasmic localization (bottom number). MCF-7 control cells show a characteristic epithelial phenotype with high E-cadherin, low vimentin, low keratin 5/6 expression and strong membrane and cytoplasmic localization of cytokeratins, MDA-231 control cells are mostly mesenchymal with low E-cadherin, high vimentin and decreased cytokeratins. Following exposure to TCR-CM, SUM149 cells show increased expression of E-cadherin, vimentin, keratin 5/6 staining and decreased pan cytokeratin staining. (c) Expression levels of EMT-related transcription factors SNAIL, ZEB1 and TG2 were quantified by Taq-Man qRT-PCR. TCR-CM and to a lesser extent LPS-CM, induced large increases in ZEB1 and TG2.
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pone.0132710.g002: Conditioned media from activated healthy donor PBMC induces EMT in IBC.(a) Morphological changes visualized by bright field in IBC cell lines consistent with stress and EMT were observed following 48-hour incubation with CM. (b) Immunohistochemical stains of paraffin cell blocks: the percentage of positive cells is listed in each image and shown in the bar graph at right. Pancytokeratin expression is shown for both percent of cells with membrane localization (top number) and cytoplasmic localization (bottom number). MCF-7 control cells show a characteristic epithelial phenotype with high E-cadherin, low vimentin, low keratin 5/6 expression and strong membrane and cytoplasmic localization of cytokeratins, MDA-231 control cells are mostly mesenchymal with low E-cadherin, high vimentin and decreased cytokeratins. Following exposure to TCR-CM, SUM149 cells show increased expression of E-cadherin, vimentin, keratin 5/6 staining and decreased pan cytokeratin staining. (c) Expression levels of EMT-related transcription factors SNAIL, ZEB1 and TG2 were quantified by Taq-Man qRT-PCR. TCR-CM and to a lesser extent LPS-CM, induced large increases in ZEB1 and TG2.

Mentions: To test the hypothesis that soluble factors secreted by activated immune cells can induce EMT in IBC cells, CM were prepared from activated immune cells and added to established 2D cultures of IBC cell lines SUM149, SUM190, IBC3 and KPL4. Cells were grown 2 days in the presence of CM prior to assaying (see Fig 1a for schema). Morphological examination under bright field revealed that cells cultured with TCR-CM and to a lesser extent, cells cultured with LPS-CM, exhibited a mesenchymal or stressed morphology with elongated projections consistent with EMT (Fig 2a).


Inflammation Mediated Metastasis: Immune Induced Epithelial-To-Mesenchymal Transition in Inflammatory Breast Cancer Cells.

Cohen EN, Gao H, Anfossi S, Mego M, Reddy NG, Debeb B, Giordano A, Tin S, Wu Q, Garza RJ, Cristofanilli M, Mani SA, Croix DA, Ueno NT, Woodward WA, Luthra R, Krishnamurthy S, Reuben JM - PLoS ONE (2015)

Conditioned media from activated healthy donor PBMC induces EMT in IBC.(a) Morphological changes visualized by bright field in IBC cell lines consistent with stress and EMT were observed following 48-hour incubation with CM. (b) Immunohistochemical stains of paraffin cell blocks: the percentage of positive cells is listed in each image and shown in the bar graph at right. Pancytokeratin expression is shown for both percent of cells with membrane localization (top number) and cytoplasmic localization (bottom number). MCF-7 control cells show a characteristic epithelial phenotype with high E-cadherin, low vimentin, low keratin 5/6 expression and strong membrane and cytoplasmic localization of cytokeratins, MDA-231 control cells are mostly mesenchymal with low E-cadherin, high vimentin and decreased cytokeratins. Following exposure to TCR-CM, SUM149 cells show increased expression of E-cadherin, vimentin, keratin 5/6 staining and decreased pan cytokeratin staining. (c) Expression levels of EMT-related transcription factors SNAIL, ZEB1 and TG2 were quantified by Taq-Man qRT-PCR. TCR-CM and to a lesser extent LPS-CM, induced large increases in ZEB1 and TG2.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4514595&req=5

pone.0132710.g002: Conditioned media from activated healthy donor PBMC induces EMT in IBC.(a) Morphological changes visualized by bright field in IBC cell lines consistent with stress and EMT were observed following 48-hour incubation with CM. (b) Immunohistochemical stains of paraffin cell blocks: the percentage of positive cells is listed in each image and shown in the bar graph at right. Pancytokeratin expression is shown for both percent of cells with membrane localization (top number) and cytoplasmic localization (bottom number). MCF-7 control cells show a characteristic epithelial phenotype with high E-cadherin, low vimentin, low keratin 5/6 expression and strong membrane and cytoplasmic localization of cytokeratins, MDA-231 control cells are mostly mesenchymal with low E-cadherin, high vimentin and decreased cytokeratins. Following exposure to TCR-CM, SUM149 cells show increased expression of E-cadherin, vimentin, keratin 5/6 staining and decreased pan cytokeratin staining. (c) Expression levels of EMT-related transcription factors SNAIL, ZEB1 and TG2 were quantified by Taq-Man qRT-PCR. TCR-CM and to a lesser extent LPS-CM, induced large increases in ZEB1 and TG2.
Mentions: To test the hypothesis that soluble factors secreted by activated immune cells can induce EMT in IBC cells, CM were prepared from activated immune cells and added to established 2D cultures of IBC cell lines SUM149, SUM190, IBC3 and KPL4. Cells were grown 2 days in the presence of CM prior to assaying (see Fig 1a for schema). Morphological examination under bright field revealed that cells cultured with TCR-CM and to a lesser extent, cells cultured with LPS-CM, exhibited a mesenchymal or stressed morphology with elongated projections consistent with EMT (Fig 2a).

Bottom Line: It is unknown whether immune cells associated to the IBC microenvironment play a role in this scenario to transiently promote epithelial to mesenchymal transition (EMT) in these cells.Interestingly, although IBC cells exhibited increased invasion and migration following exposure to immune factors, the expression of E-cadherin (CDH1), a cell adhesion molecule, increased uniquely in IBC cell lines but not in non-IBC cell lines.These data suggest that release of cytokines by activated immune cells may contribute to the aggressiveness of IBC and highlight these factors as potential target mediators of immune-IBC interaction.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; The Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; The University of Texas Graduate School of Biomedical Sciences at Houston, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.

ABSTRACT
Inflammatory breast cancer (IBC) is the most insidious form of locally advanced breast cancer; about a third of patients have distant metastasis at initial staging. Emerging evidence suggests that host factors in the tumor microenvironment may interact with underlying IBC cells to make them aggressive. It is unknown whether immune cells associated to the IBC microenvironment play a role in this scenario to transiently promote epithelial to mesenchymal transition (EMT) in these cells. We hypothesized that soluble factors secreted by activated immune cells can induce an EMT in IBC and thus promote metastasis. In a pilot study of 16 breast cancer patients, TNF-α production by peripheral blood T cells was correlated with the detection of circulating tumor cells expressing EMT markers. In a variety of IBC model cell lines, soluble factors from activated T cells induced expression of EMT-related genes, including FN1, VIM, TGM2, ZEB1. Interestingly, although IBC cells exhibited increased invasion and migration following exposure to immune factors, the expression of E-cadherin (CDH1), a cell adhesion molecule, increased uniquely in IBC cell lines but not in non-IBC cell lines. A combination of TNF-α, IL-6, and TGF-β was able to recapitulate EMT induction in IBC, and conditioned media preloaded with neutralizing antibodies against these factors exhibited decreased EMT. These data suggest that release of cytokines by activated immune cells may contribute to the aggressiveness of IBC and highlight these factors as potential target mediators of immune-IBC interaction.

No MeSH data available.


Related in: MedlinePlus