Cyclin-dependent kinase inhibitors and basement membrane interact to regulate breast epithelial cell differentiation and acinar morphogenesis.

2.50
Hdl Handle:
http://hdl.handle.net/10541/70473
Title:
Cyclin-dependent kinase inhibitors and basement membrane interact to regulate breast epithelial cell differentiation and acinar morphogenesis.
Authors:
Coppock, H A; Gilham, David E; Howell, Anthony ( 0000-0002-3879-5991 ) ; Clarke, Robert B
Abstract:
OBJECTIVE: The cyclin-dependent kinase inhibitors (CDKIs), p21(CIP1) and p27(KIP1) regulate growth and differentiation in diverse tissue types. We aimed to determine whether p21(CIP1) or p27(KIP1) could induce a terminally differentiated phenotype in breast cells, and to examine if CDKI expression is regulated by basement membrane interactions. MATERIALS AND METHODS: Effects of increased CDKI expression on the phenotype of MCF-10A breast epithelial cells were examined by retroviral transduction of p21(CIP1) or p27(KIP1) cDNA. RESULTS: Overexpression of p21(CIP1) or p27(KIP1) reduced MCF-10A growth rates in monolayer cultures, altered cellular morphology and stimulated accumulation of neutral lipid droplets, suggesting partial lactational differentiation. However, markers of luminal differentiation (oestrogen and progesterone receptors, alpha-lactalbumin, beta-casein and adipophilin) were absent when examined by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Cell-basement membrane contacts are known to be essential for full mammary epithelial cell differentiation and therefore parental MCF-10A cells were cultured on a basement membrane preparation (Matrigel) in which they form acini. Immunocytochemistry showed that Ki67, the cell proliferation marker, was initially expressed at high levels and as growth decreased p27(KIP1) expression steadily increased. Surprisingly, p21(CIP1) was highest at the early stages of acinus growth and was detected in proliferating cells, as demonstrated by colocalization in dual Ki67/p21(CIP1) immunofluorescence. Overexpression of p21(CIP1) or p27(KIP1) impaired formation of acini, whereas their knockdown, using siRNA, increased acinus formation. CONCLUSION: We conclude that both p21(CIP1) and p27(KIP1) induce partial secretory differentiation of mammary cells in monolayer, but during acinus morphogenesis in 3D culture they have a highly regulated temporal expression pattern.
Affiliation:
Centre for Molecular Medicine, University of Manchester, Manchester, UK.
Citation:
Cyclin-dependent kinase inhibitors and basement membrane interact to regulate breast epithelial cell differentiation and acinar morphogenesis. 2007, 40 (5):721-40 Cell Prolif.
Journal:
Cell Proliferation
Issue Date:
Oct-2007
URI:
http://hdl.handle.net/10541/70473
DOI:
10.1111/j.1365-2184.2007.00463.x
PubMed ID:
17877612
Type:
Article
Language:
en
ISSN:
0960-7722
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorCoppock, H A-
dc.contributor.authorGilham, David E-
dc.contributor.authorHowell, Anthony-
dc.contributor.authorClarke, Robert B-
dc.date.accessioned2009-06-15T12:03:22Z-
dc.date.available2009-06-15T12:03:22Z-
dc.date.issued2007-10-
dc.identifier.citationCyclin-dependent kinase inhibitors and basement membrane interact to regulate breast epithelial cell differentiation and acinar morphogenesis. 2007, 40 (5):721-40 Cell Prolif.en
dc.identifier.issn0960-7722-
dc.identifier.pmid17877612-
dc.identifier.doi10.1111/j.1365-2184.2007.00463.x-
dc.identifier.urihttp://hdl.handle.net/10541/70473-
dc.description.abstractOBJECTIVE: The cyclin-dependent kinase inhibitors (CDKIs), p21(CIP1) and p27(KIP1) regulate growth and differentiation in diverse tissue types. We aimed to determine whether p21(CIP1) or p27(KIP1) could induce a terminally differentiated phenotype in breast cells, and to examine if CDKI expression is regulated by basement membrane interactions. MATERIALS AND METHODS: Effects of increased CDKI expression on the phenotype of MCF-10A breast epithelial cells were examined by retroviral transduction of p21(CIP1) or p27(KIP1) cDNA. RESULTS: Overexpression of p21(CIP1) or p27(KIP1) reduced MCF-10A growth rates in monolayer cultures, altered cellular morphology and stimulated accumulation of neutral lipid droplets, suggesting partial lactational differentiation. However, markers of luminal differentiation (oestrogen and progesterone receptors, alpha-lactalbumin, beta-casein and adipophilin) were absent when examined by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Cell-basement membrane contacts are known to be essential for full mammary epithelial cell differentiation and therefore parental MCF-10A cells were cultured on a basement membrane preparation (Matrigel) in which they form acini. Immunocytochemistry showed that Ki67, the cell proliferation marker, was initially expressed at high levels and as growth decreased p27(KIP1) expression steadily increased. Surprisingly, p21(CIP1) was highest at the early stages of acinus growth and was detected in proliferating cells, as demonstrated by colocalization in dual Ki67/p21(CIP1) immunofluorescence. Overexpression of p21(CIP1) or p27(KIP1) impaired formation of acini, whereas their knockdown, using siRNA, increased acinus formation. CONCLUSION: We conclude that both p21(CIP1) and p27(KIP1) induce partial secretory differentiation of mammary cells in monolayer, but during acinus morphogenesis in 3D culture they have a highly regulated temporal expression pattern.en
dc.language.isoenen
dc.subject.meshBase Sequence-
dc.subject.meshBasement Membrane-
dc.subject.meshBreast-
dc.subject.meshCell Differentiation-
dc.subject.meshCell Line-
dc.subject.meshCyclin-Dependent Kinase Inhibitor Proteins-
dc.subject.meshCyclin-Dependent Kinase Inhibitor p21-
dc.subject.meshEpithelial Cells-
dc.subject.meshFemale-
dc.subject.meshHumans-
dc.subject.meshIntracellular Signaling Peptides and Proteins-
dc.subject.meshKi-67 Antigen-
dc.subject.meshLipid Metabolism-
dc.subject.meshMorphogenesis-
dc.subject.meshRNA, Small Interfering-
dc.subject.meshTransduction, Genetic-
dc.titleCyclin-dependent kinase inhibitors and basement membrane interact to regulate breast epithelial cell differentiation and acinar morphogenesis.en
dc.typeArticleen
dc.contributor.departmentCentre for Molecular Medicine, University of Manchester, Manchester, UK.en
dc.identifier.journalCell Proliferationen
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