Comparative Proteomic Analysis of Proliferating and Functionally Differentiated Mammary Epithelial Cells

Desrivières, Sylvane; Prinz, Thorsten; Laria, Nahomi Castro-Palomino; Meyer, Markus; Boehm, Gitte; Bauer, Ute; Schäfer, Jürgen; Neumann, Thomas; Shemanko, Carrie S.; Groner, Bernd

doi:10.1074/mcp.m300032-mcp200

Cited by 59 publications

(57 citation statements)

References 68 publications

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“…has been shown to be upregulated in mammary-gland cells as a result of lactogenic hormone-stimulated differentiation (Desrivieres et al, 2003), so downregulation of Tcp1 might also account for the less differentiated phenotype that we observe in miR-205-overexpressing cells.…”

Section: Journal Of Cell Science 123 (4)mentioning

confidence: 53%

A putative role for microRNA-205 in mammary epithelial cell progenitors

Greene

Gunaratne

Hammond

et al. 2010

Journal of Cell Science

124

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SummaryIn an effort to understand the potential role of microRNAs (miRNAs) in mammary-gland stem or progenitor cells, miRNA microarrays were performed on subpopulations of the mouse mammary epithelial cell (MEC) line COMMA-DbGeo. This cell line contains a heterogeneous subpopulation of progenitors characterized by the expression of stem cell antigen 1 (Sca-1; encoded by Ly6a). Microarray analysis indicated that the Sca-1 subpopulations have distinct miRNA expression profiles. Functional studies were performed on miR-205, which was highly expressed in the Sca-1-positive (Sca-1 + ) cells. When miR-205 was overexpressed in vitro, the COMMA-DbGeo cells underwent several significant morphological and molecular changes. miR-205 overexpression led to an expansion of the progenitorcell population, decreased cell size and increased cellular proliferation. In addition, the colony-forming potential of the two Sca-1 subpopulations was increased. Target prediction for miR-205 indicated that it might regulate the expression of the tumor-suppressor protein PTEN. Overexpression studies using reporter constructs confirmed that PTEN expression is regulated by miR-205. In addition to PTEN, several other putative and previously validated miR-205 targets were identified by microarray analysis, including the previously reported miR-205 targets ZEB1 and ZEB2. Additionally, in normal mouse MECs, high expression of miR-205 was observed in stemcell-enriched cell populations isolated by FACS using established cell-surface markers.

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Section: Journal Of Cell Science 123 (4)mentioning

confidence: 53%

A putative role for microRNA-205 in mammary epithelial cell progenitors

Greene

Gunaratne

Hammond

et al. 2010

Journal of Cell Science

124

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“…To assess the potential role of Stim2 in mammary epithelial cells during lactation, we used the HC11 mouse mammary epithelial cell line model (36). Differentiation of HC11 cells using lactogenic hormones significantly induced the mRNA expression of the milk protein b-casein (data not shown), which was undetectable prior to differentiation (16,37). Treatment of differentiated HC11 cells with Stim2 siRNA caused a significant decrease in basal [Ca 2þ ] I (Fig.…”

Section: Orai1 Calcium Channel Modulators Stim1 and Stim2 During Mammmentioning

confidence: 99%

ORAI1-Mediated Calcium Influx in Lactation and in Breast Cancer

McAndrew

Grice

Peters

et al. 2011

Molecular Cancer Therapeutics

196

212

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The entry of calcium into the mammary epithelial cell from the maternal plasma (i.e., calcium influx mechanisms) during lactation is poorly understood. As alterations in calcium channels and pumps are a key feature of some cancers, including breast cancer, understanding these calcium influx pathways may have significance beyond mammary biology. We show that the store-operated calcium influx protein, Orai1, is increased during lactation whereas the Orai1 activator Stim1, but not Stim2, is downregulated. Stim2 siRNA reduced basal calcium levels in a lactation model. Our results suggest that calcium influx is remodeled in mammary epithelial cells during lactation, with calcium influx increased through Orai1, activated by Stim2. Breast cancer cell lines had increased levels of ORAI1. ORAI1 siRNA in breast cancer cells reduced storeoperated calcium entry and remodeled the calcium influx associated with invasive stimuli. Analysis of microarray data from 295 breast cancers showed that the transcriptional breast cancer subtype with the poorest prognosis (basal) was associated with an altered relationship between the ORAI1 regulators STIM1 and STIM2, and that women with breast cancers with STIM1 high /STIM2 low tumors had a significantly poorer prognosis. Our studies show that during lactation there is a remodeling in the nature of calcium influx and that alteration in the ORAI1 influx pathway may be a feature of some breast cancers, particularly those with the poorest prognosis. Our studies suggest that this pathway may be a novel therapeutic target for breast cancer treatment in these women. Mol Cancer Ther; 10(3); 448-60. Ó2011 AACR.

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“…The enzyme glucose-6-phosphate dehydrogenase decreased in abundance, suggesting a decrease in the amount of glucose used for NADPH synthesis, sparing glucose for lactose synthesis. To prepare for lactation, MECs must also undergo polarization through reorganization of the cytoskeleton for milk synthesis and secretion (Desrivieres et al, 2003, Wodarz, 2002. Multiple proteins involved in actin cytoskeletal rearrangement also showed significant changes in response to GH in the present study.…”

Section: Discussionsupporting

confidence: 51%

“…In vitro, MECs are commonly differentiated with lactogenic hormones, dexamethasone, insulin and prolactin (DIP) (Desrivieres et al, 2003, Matitashvili et al, 1997, Topper and Freeman, 1980, Zhou et al, 2008a. Differentiation has also been attributed to GH and the transcription factor STAT5 and is commonly measured by milk protein mRNA expression.…”

Section: Mammary Gland Developmentmentioning

confidence: 99%

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Growth hormone alters components related to differentiation, metabolism and milk synthesis and secretion in MAC-T cells

Johnson¹

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iv GROWTH HORMONE ALTERS COMPONENTS RELATED TO DIFFERENTIATION, METABOLISM AND MILK SYNTHESIS AND SECRETION IN MAC-T CELLS Tasha L. JohnsonThe mammary alveolar cell-T (MAC-T) cell line is able to uniformly differentiate and secrete casein proteins in response to dexamethasone, insulin and prolactin and is extensively used to study bovine mammary epithelial cell function. Growth hormone (GH) has been shown to increase milk protein synthesis both in vivo and in mammary cell models, and induce cytoskeletal rearrangement in 3T3 fibroblast cell line and a Chinese hamster ovary (CHO) cell line. Few studies have focused on identifying the mechanisms involved in differentiated MAC-T cells' response to GH. We tested the hypothesis that MAC-T cells would respond directly to GH and that the response would include alterations in milk protein gene expression, leading to a more appropriate model for mammary cell function than treatment with dexamethasone, insulin and prolactin alone. To identify mechanisms that are involved in MAC-T cells' response to GH, global protein was assessed through two-dimensional gel electrophoresis and differentially expressed proteins were identified through mass spectrometry. Differentiated cells expressed GH receptor mRNA, and addition of GH to the differentiation medium increased production of α-S1 casein and α-lactalbumin mRNA. Proteins that were differentially expressed are related to metabolism, the cytoskeleton, protein folding, RNA and DNA processing, detoxifying and calcium metabolism. These results indicate that GH is an important factor in inducing a lactogenic phenotype in the MAC-T cell line, and supports GHs involvement in differentiation, while altering cell metabolism in preparation for synthesis and secretion of milk components. v ACKNOWLEDGMENTS

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Comparative Proteomic Analysis of Proliferating and Functionally Differentiated Mammary Epithelial Cells

Cited by 59 publications

References 68 publications

A putative role for microRNA-205 in mammary epithelial cell progenitors

A putative role for microRNA-205 in mammary epithelial cell progenitors

ORAI1-Mediated Calcium Influx in Lactation and in Breast Cancer

Growth hormone alters components related to differentiation, metabolism and milk synthesis and secretion in MAC-T cells

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