Abstract
Mammary epithelial cells cultured on a concentrated laminin-rich extracellular matrix formed 3D acinar structures that matured to polarized monolayers surrounding a lumen. In the absence of glucocorticoids mature acinus formation failed and the expression of an acinus-associated, activator protein 1 (AP1) and nuclear factor kappaB transcription factor DNA-binding profile was lost. Treatment with the JNK inhibitor, SP600125, caused similar effects, whereas normal organization of the mammary epithelial cells as acini caused JNK activation in a glucocorticoid-dependent manner. The forming acini expressed BRCA1, GADD45beta, MEKK4, and the JNK activating complex GADD 45beta-MEKK4 in a glucocorticoid-dependent fashion. JNK catalyzed phosphorylation of c-Jun was also detected in the acini. In addition, expression of beta4 integrin and in situ occupation of its promoter by AP1 components, c-Jun and Fos, was glucocorticoid dependent. These results suggest that glucocortocoid signaling regulates acinar integrity through a pathway involving JNK regulation of AP1 transcription factors and beta4 integrin expression.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Anthracenes / pharmacology
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BRCA1 Protein / metabolism
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Blotting, Western
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Cell Culture Techniques / methods
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Cells, Cultured
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Chromatin / metabolism
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Epithelial Cells / cytology*
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Extracellular Matrix / metabolism
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Glucocorticoids / metabolism*
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Integrin beta4 / metabolism
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Integrins / metabolism
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JNK Mitogen-Activated Protein Kinases*
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Lactation
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Luciferases / metabolism
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MAP Kinase Kinase 4
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Mammary Glands, Animal / cytology*
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Mice
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Microscopy, Fluorescence
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Mitogen-Activated Protein Kinase Kinases / metabolism*
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Models, Biological
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Phosphorylation
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Precipitin Tests
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Proto-Oncogene Proteins c-fos / metabolism
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Proto-Oncogene Proteins c-jun / metabolism
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RNA, Messenger / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Signal Transduction
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Time Factors
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Transcription Factor AP-1 / metabolism
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Transfection
Substances
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Anthracenes
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BRCA1 Protein
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Chromatin
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Glucocorticoids
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Integrin beta4
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Integrins
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Proto-Oncogene Proteins c-fos
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Proto-Oncogene Proteins c-jun
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RNA, Messenger
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Transcription Factor AP-1
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pyrazolanthrone
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Luciferases
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JNK Mitogen-Activated Protein Kinases
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MAP Kinase Kinase 4
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Mitogen-Activated Protein Kinase Kinases