Document Detail

Essential role of biliary glycoprotein (CD66a) in morphogenesis of the human mammary epithelial cell line MCF10F.
MedLine Citation:
PMID:  10564638     Owner:  NLM     Status:  MEDLINE    
Normal mammary epithelial cells express the cell surface protein biliary glycoprotein (BGP or CD66a) in a polarized manner, suggesting that this protein may play a role in the formation of mammary acini. In order to test this hypothesis, we interrupted the expression of BGP in the mammary epithelial line MCF10F when cultured in or on Matrigel, a source of extracellular matrix (ECM). When analyzed by immunofluorescence confocal microscopy, the BGP staining is confined to the lumenal surface and colocalizes with actin. Sequential scanning electron microscopy demonstrates that the MCF10F cells migrate to form clusters, followed by apoptotic cell death within the center, resulting in lumen formation. Transmission electron micrographs reveal the presence of tight junctions and desmosomes between the cells, microvilli along the lumenal surface, and typical apoptotic bodies within the lumen. When the MCF10F cells are transfected with the BGP antisense gene and grown in Matrigel, they exhibit reduced acini formation (12% and 20%) compared to untransfected cells (52%) or to cells transfected with vector only (62%). Acini formation is also significantly reduced when MCF10F cells grown in Matrigel are treated with anti-BGP antibody (18% at 100 microgram/ml), or recombinant soluble BGP (18% at 0.4 microM). In contrast, the BGP-negative MCF7 breast tumor cell line, which does not form acini when grown in matrigel, exhibits >60% cell death with the occasional formation of acini, when transfected with the BGP sense gene and grown in Matrigel. These results support the hypothesis that BGP plays a role in the normal differentiation program of mammary epithelial cells, indicating that its expression is essential to the formation of the lumen. Furthermore, and as shown by others, the differentiation program depends on the presence of ECM. The lack of expression of BGP in the MCF7 breast cancer cell line suggests that the downregulation of BGP expression confers a growth advantage to these cells in ECM. In addition, we found that the MCF10F cells could be separated into a BGP-positive epithelial fraction (MCF10F-e), and a BGP-negative myoepithelial fraction (MCF10F-m). When the myoepithelial cell-enriched fraction is grown on Matrigel, web-like structures are formed. These cells have a typical spindle shape cell morphology and express keratin, alpha-smooth muscle actin and vimentin, markers of the myoepithelial cell phenotype. When MCF10F-m cells are treated with IFNgamma, they express CEA (carcinoembryonic antigen) but not BGP. Since breast carcinomas, especially in situ carcinomas, express CEA, this finding may suggest a heretofore unappreciated relationship between myoepithelial cells and breast cancer.
J Huang; J D Hardy; Y Sun; J E Shively
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of cell science     Volume:  112 ( Pt 23)     ISSN:  0021-9533     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  1999 Dec 
Date Detail:
Created Date:  2000-02-03     Completed Date:  2000-02-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  4193-205     Citation Subset:  IM    
Graduate School of the City of Hope, Beckman Research Institute, Duarte, CA 91010, USA.
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MeSH Terms
Antigens, CD / genetics*,  physiology
Antigens, Differentiation / genetics*,  physiology
Breast Neoplasms
Cell Adhesion / physiology
Cell Adhesion Molecules / physiology
Culture Media
Epithelial Cells / cytology,  physiology,  ultrastructure
Gene Expression Regulation
Microscopy, Electron, Scanning
Morphogenesis / physiology*
Recombinant Proteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Tumor Cells, Cultured
Grant Support
Reg. No./Substance:
0/Antigens, CD; 0/Antigens, Differentiation; 0/CD66 antigens; 0/Cell Adhesion Molecules; 0/Culture Media; 0/Recombinant Proteins

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