Journal of Cell Science, Vol 107, Issue 10 2931-2939, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Ectoenzyme regulation by phenotypically distinct fibroblast sub-populations isolated from the human mammary gland

AJ Atherton, MJ O'Hare, L Buluwela, J Titley, P Monaghan, HF Paterson, MJ Warburton and BA Gusterson
Institute of Cancer Research, Haddow Laboratories, Sutton, Surrey, UK.

Inter- and intralobular mammary fibroblasts have been separated from normal human breast tissue and cultured to study the differential expression of ectoenzymes present within the stroma of the normal gland and associated with breast cancers. Specific ectoenzymes were identified by indirect immunofluorescence and quantified by flow cytometry and semi-quantitative PCR. A consistent difference was noted between the two fibroblast sub-populations at early passage in respect of dipeptidyl peptidase IV (DPP IV) and aminopeptidase N (APN) expression. Early passage intralobular fibroblasts were positive for APN but negative for DPP IV, as seen in the intact tissue. However, with continued sub-culture they gradually began to express DPP IV, until at later passages they became indistinguishable from the interlobular fibroblasts, which were APN and DPP IV-positive at all stages in culture, as they are in intact tissue. Neutral endopeptidase (NEP/CALLA/CD10) is not expressed by normal adult breast fibroblasts but is found in the stroma associated with over 60% of breast cancers. It was up-regulated in vitro on both inter- and intralobular fibroblasts, with final levels that were significantly (< 14 times) higher on the former in all pairs of preparations from individual donors analysed. This difference persisted with continued passage, and levels of the ectoenzyme and its messenger RNA were further up-regulated by hydrocortisone in both populations. These results demonstrate that phenotypically distinct cultures of human mammary fibroblast sub-populations can be used to study the regulation of these stromal ectoenzymes.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

				Y. Sun, K. Boyd, W. Xu, J. Ma, C. W. Jackson, A. Fu, J. M. Shillingford, G. W. Robinson, L. Hennighausen, J. K. Hitzler, et al. Acute Myeloid Leukemia-Associated Mkl1 (Mrtf-a) Is a Key Regulator of Mammary Gland Function Mol. Cell. Biol., August 1, 2006; 26(15): 5809 - 5826. [Abstract] [Full Text] [PDF]

				S. J. Mabjeesh, O. Gal-Garber, J. Milgram, Y. Feuermann, M. Cohen-Zinder, and A. Shamay Aminopeptidase N Gene Expression and Abundance in Caprine Mammary Gland is Influenced by Circulating Plasma Peptide J Dairy Sci, June 1, 2005; 88(6): 2055 - 2064. [Abstract] [Full Text] [PDF]

				A N Kalof, D Tam, B Beatty, and K Cooper Immunostaining patterns of myoepithelial cells in breast lesions: a comparison of CD10 and smooth muscle myosin heavy chain J. Clin. Pathol., June 1, 2004; 57(6): 625 - 629. [Abstract] [Full Text] [PDF]

				M. J. O'Hare, J. Bond, C. Clarke, Y. Takeuchi, A. J. Atherton, C. Berry, J. Moody, A. R. J. Silver, D. C. Davies, A. E. Alsop, et al. Conditional immortalization of freshly isolated human mammary fibroblasts and endothelial cells PNAS, January 16, 2001; 98(2): 646 - 651. [Abstract] [Full Text] [PDF]

				D. Kurz, S Decary, Y Hong, and J. Erusalimsky Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells J. Cell Sci., January 10, 2000; 113(20): 3613 - 3622. [Abstract] [PDF]

				G. Cutrona, N. Leanza, M. Ulivi, G. Melioli, V. L. Burgio, G. Mazzarello, G. Gabutti, S. Roncella, and M. Ferrarini Expression of CD10 by Human T Cells That Undergo Apoptosis Both In Vitro and In Vivo Blood, November 1, 1999; 94(9): 3067 - 3076. [Abstract] [Full Text] [PDF]

				M. J. Page, B. Amess, R. R. Townsend, R. Parekh, A. Herath, L. Brusten, M. J. Zvelebil, R. C. Stein, M. D. Waterfield, S. C. Davies, et al. Proteomic definition of normal human luminal and myoepithelial breast cells purified from reduction mammoplasties PNAS, October 26, 1999; 96(22): 12589 - 12594. [Abstract] [Full Text] [PDF]

				L. Kuru, M.H. Parkar, G.S. Griffiths, H.N. Newman, and I. Olsen Flow Cytometry Analysis of Gingival and Periodontal Ligament Cells Journal of Dental Research, April 1, 1998; 77(4): 555 - 564. [Abstract] [PDF]

				V Bender, S Buschlen, and D Cassio Expression and localization of hepatocyte domain-specific plasma membrane proteins in hepatoma x fibroblast hybrids and in hepatoma dedifferentiated variants J. Cell Sci., January 11, 1998; 111(22): 3437 - 3450. [Abstract] [PDF]

				B Niranjan, L Buluwela, J Yant, N Perusinghe, A Atherton, D Phippard, T Dale, B Gusterson, and T Kamalati HGF/SF: a potent cytokine for mammary growth, morphogenesis and development Development, January 9, 1995; 121(9): 2897 - 2908. [Abstract] [PDF]