Nidogen-1 regulates laminin-1-dependent mammary-specific gene expression

Nidogen-1 (entactin) acts as a bridge between the extracellular matrix molecules laminin-1 and type IV collagen, and thus participates in the assembly of basement membranes. To investigate the role of nidogen-1 in regulating cell-type-specific gene expression in mammary epithelium, we designed a culture microecosystem in which each component, including epithelial cells, mesenchymal cells, lactogenic hormones and extracellular matrix, could be controlled. We found that primary and established mesenchymal and myoepithelial cells synthesized and secreted nidogen-1, whereas expression was absent in primary and established epithelial cells. In an epithelial cell line containing mesenchymal cells, nidogen-1 was produced by the mesenchymal cells but deposited between the epithelial cells. In this mixed culture, mammary epithelial cells express (beta)-casein in the presence of lactogenic hormones. Addition of either laminin-1 plus nidogen-1, or laminin-1 alone, to mammary epithelial cells induced (beta)-casein production. We asked whether recombinant nidogen-1 alone could signal directly for (beta)-casein. Nidogen-1 did not induce (beta)-casein synthesis in epithelial cells, but it augmented the inductive capacity of laminin-1. These data suggest that nidogen-1 can cooperate with laminin-1 to regulate (beta)-casein expression. Addition of full-length nidogen-1 to the mixed cultures had no effect on (beta)-casein gene expression; however, a nidogen-1 fragment containing the laminin-1 binding domain, but lacking the type IV collagen-binding domain, had a dominant negative effect on (beta)-casein expression. These data point to a physiological role for nidogen-1 in the basement membrane-induced gene expression by epithelial cells.

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A Dual Role for Oncostatin M Signaling in the Differentiation and Death of Mammary Epithelial Cells in Vivo

Molecular Endocrinology ◽

10.1210/me.2008-0097 ◽

2008 ◽

Vol 22 (12) ◽

pp. 2677-2688 ◽

Cited By ~ 45

Author(s):

Paul G. Tiffen ◽

Nader Omidvar ◽

Nuria Marquez-Almuina ◽

Dawn Croston ◽

Christine J. Watson ◽

...

Keyword(s):

Gene Expression ◽

Epithelial Cell ◽

Epithelial Cells ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Oncostatin M ◽

Specific Gene ◽

Mammary Involution

Abstract Recent studies in breast cancer cell lines have shown that oncostatin M (OSM) not only inhibits proliferation but also promotes cell detachment and enhances cell motility. In this study, we have looked at the role of OSM signaling in nontransformed mouse mammary epithelial cells in vitro using the KIM-2 mammary epithelial cell line and in vivo using OSM receptor (OSMR)-deficient mice. OSM and its receptor were up-regulated approximately 2 d after the onset of postlactational mammary regression, in response to leukemia inhibitory factor (LIF)-induced signal transducer and activator of transcription-3 (STAT3). This resulted in sustained STAT3 activity, increased epithelial apoptosis, and enhanced clearance of epithelial structures during the remodeling phase of mammary involution. Concurrently, OSM signaling precipitated the dephosphorylation of STAT5 and repressed expression of the milk protein genes β-casein and whey acidic protein (WAP). Similarly, during pregnancy, OSM signaling suppressed β-casein and WAP gene expression. In vitro, OSM but not LIF persistently down-regulated phosphorylated (p)-STAT5, even in the continued presence of prolactin. OSM also promoted the expression of metalloproteinases MMP3, MMP12, and MMP14, which, in vitro, were responsible for OSM-specific apoptosis. Thus, the sequential activation of IL-6-related cytokines during mammary involution culminates in an OSM-dependent repression of epithelial-specific gene expression and the potentiation of epithelial cell extinction mediated, at least in part, by the reciprocal regulation of p-STAT5 and p-STAT3.

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Tenascin-C inhibits extracellular matrix-dependent gene expression in mammary epithelial cells. Localization of active regions using recombinant tenascin fragments

Journal of Cell Science ◽

10.1242/jcs.108.2.519 ◽

1995 ◽

Vol 108 (2) ◽

pp. 519-527 ◽

Cited By ~ 6

Author(s):

P.L. Jones ◽

N. Boudreau ◽

C.A. Myers ◽

H.P. Erickson ◽

M.J. Bissell

Keyword(s):

Gene Expression ◽

Extracellular Matrix ◽

Protein Synthesis ◽

Epithelial Cells ◽

Mammary Epithelial Cells ◽

Active Regions ◽

Mammary Epithelial ◽

Dependent Manner ◽

Beta Casein ◽

Casein Protein

The physiological role of tenascin in vivo has remained obscure. Although tenascin is regulated in a stage and tissue-dependent manner, knock-out mice appear normal. When tenascin expression was examined in the normal adult mouse mammary gland, little or none was present during lactation, when epithelial cells actively synthesize and secrete milk proteins in an extracellular matrix/lactogenic hormone-dependent manner. In contrast, tenascin was prominently expressed during involution, a stage characterized by the degradation of the extracellular matrix and the subsequent loss of milk production. Studies with mammary cell lines indicated that tenascin expression was high on plastic, but was suppressed in the presence of the laminin-rich, Engelbreth-Holm-Swarm (EHS) tumour biomatrix. When exogenous tenascin was added together with EHS to mammary epithelial cells, beta-casein protein synthesis and steady-state mRNA levels were inhibited in a concentration-dependent manner. Moreover, this inhibition by tenascin could be segregated from its effects on cell morphology. Using two beta-casein promoter constructs attached to the chloramphenicol acetyltransferase reporter gene we showed that tenascin selectively suppressed extracellular matrix/prolactin-dependent transcription of the beta-casein gene in three-dimensional cultures. Finally, we mapped the active regions within the fibronectin type III repeat region of the tenascin molecule that are capable of inhibiting beta-casein protein synthesis. Our data are consistent with a model where both the loss of a laminin-rich basement membrane by extracellular matrix-degrading enzymes and the induction of tenascin contribute to the loss of tissue-specific gene expression and thus the involuting process.

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Extracellular matrix-dependent tissue-specific gene expression in mammary epithelial cells requires both physical and biochemical signal transduction.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.91.26.12378 ◽

1994 ◽

Vol 91 (26) ◽

pp. 12378-12382 ◽

Cited By ~ 311

Author(s):

C. D. Roskelley ◽

P. Y. Desprez ◽

M. J. Bissell

Keyword(s):

Gene Expression ◽

Signal Transduction ◽

Extracellular Matrix ◽

Epithelial Cells ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Specific Gene ◽

Tissue Specific ◽

Tissue Specific Gene ◽

Specific Gene Expression

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Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.84.1.136 ◽

1987 ◽

Vol 84 (1) ◽

pp. 136-140 ◽

Cited By ~ 312

Author(s):

M. L. Li ◽

J. Aggeler ◽

D. A. Farson ◽

C. Hatier ◽

J. Hassell ◽

...

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Basement Membrane ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Casein Gene

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Cyclooxygenase-2 Gene Expression is Upregulated in Transformed Mammary Epithelial Cells

Annals of the New York Academy of Sciences ◽

10.1111/j.1749-6632.1997.tb48606.x ◽

1997 ◽

Vol 833 (1 Cancer) ◽

pp. 179-185 ◽

Cited By ~ 9

Author(s):

KOTHA SUBBARAMAIAH ◽

NITIN TELANG ◽

MEENA B. BANSAL ◽

BABETTE B. WEKSLER ◽

ANDREW J. DANNENBERG

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Mammary Epithelial Cells ◽

Cyclooxygenase 2 ◽

Mammary Epithelial

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PSXII-17 Supplementing ten essential amino acids alters beta-casein expression and cell and medium protein synthesis in bovine mammary epithelial cells.

Journal of Animal Science ◽

10.1093/jas/sky404.581 ◽

2018 ◽

Vol 96 (suppl_3) ◽

pp. 265-265

Author(s):

J Conejos ◽

J Kim ◽

T Yadana Moe ◽

W Choe ◽

M Bae ◽

...

Keyword(s):

Amino Acids ◽

Protein Synthesis ◽

Epithelial Cells ◽

Mammary Epithelial Cells ◽

Essential Amino Acids ◽

Mammary Epithelial ◽

Bovine Mammary Epithelial Cells ◽

Beta Casein

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Gene expression of six major milk proteins in primary bovine mammary epithelial cells isolated from milk during the first twenty weeks of lactation

Czech Journal of Animal Science ◽

10.17221/6347-cjas ◽

2012 ◽

Vol 57 (No. 10) ◽

pp. 469-480 ◽

Cited By ~ 15

Author(s):

T. Sigl ◽

H.H.D. Meyer ◽

S. Wiedemann

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Milk Protein ◽

Protein Gene ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Milk Protein Gene ◽

Bovine Mammary Epithelial Cells ◽

Milk Protein Genes ◽

Milk Protein Gene Expression

The objective of the present study was to refine a previously developed method to isolate primary bovine mammary epithelial cells (pBMEC) from fresh milk. Using this method, it was tested whether the number of pBMEC and the relation of recovered pBMEC to total somatic cell count vary within the individual lactation stages. Furthermore, the expression levels of the milk protein genes during the first twenty weeks of lactation were determined by quantitative PCR method. A total number of 152 morning milk samples were obtained from twenty-four Holstein-Friesian cows during the first 20 weeks of lactation (day 8, 15, 26, 43, 57, 113, and 141 postpartum). Numbers of extracted pBMEC were consistent at all time-points (1.1 ± 0.06 to 1.4 ± 0.03 ×103/ml) and an average value of RNA integrity number (RIN) was 6.3 ± 0.3. Percentage of pBMEC in relation to total milk cells (2.0 ± 0.2 to 6.7 ± 1.0%) correlated with milk yield. Expression patterns of the casein genes alpha (α)S1, (α)S2, beta (β), and kappa (κ) (CSN1S1, CSN1S2, CSN2, CSN3, respectively) and the whey protein genes α-lactalbumin (LALBA) and progestagen-associated endometrial protein (PAEP; known as β-lactoglobulin) were shown to be comparable, i.e. transcripts of all six milk protein genes were found to peak during the first two weeks of lactation and to decline continuously towards mid lactation. However, mRNA levels were different among genes with CSN3 showing the highest and LALBA the lowest abundance. We hypothesized that milk protein gene expression has a pivotal effect on milk protein composition with no influence on milk protein concentration. This paper is the first to describe milk protein gene expression during lactation in pBMEC collected in milk. Future studies will be needed to understand molecular mechanisms in pBMEC including regulation of expression and translation throughout lactation.

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