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Proc Natl Acad Sci U S A. Oct 1, 1992; 89(19): 9064–9068.
PMCID: PMC50065

Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells.

Abstract

Normal human breast epithelial cells show a high degree of phenotypic plasticity in monolayer culture and express many traits that otherwise characterize tumor cells in vivo. Paradoxically, primary human breast carcinoma cells are difficult to establish in culture: most outgrowths arise from the normal tissue surrounding the tumor. These characteristics have posed major obstacles to the establishment of simple reliable criteria for mammary epithelial transformation in culture. In the present study, we show that a reconstituted basement membrane (BM) can be used to culture all normal human breast epithelial cells and a subset of human breast carcinoma cells. The two cell types can be readily distinguished by virtue of the ability of normal cells to reexpress a structurally and functionally differentiated phenotype within BM. Twelve specimens of normal breast tissue and 2 normal breast epithelial cell lines (total 14 samples) embedded in BM as single cells were able to form multicellular spherical colonies with a final size close to that of true acini in situ. Sections of mature spheres revealed a central lumen surrounded by polarized luminal epithelial cells expressing keratins 18 and 19 and sialomucin at the apical membrane. Significantly, two-thirds of normal spheres deposited a visible endogenous type IV collagen-containing BM even though they were in contact with exogenously provided BM. Growth was arrested completely within the same time period. In contrast, none of 6 carcinoma cell lines or 2 cultures of carcinoma from fresh samples (total 8 samples) responded to BM by growth regulation, lumen formation, correct polarity, or deposition of endogenous BM. These findings may provide the basis of a rapid assay for discriminating normal human breast epithelial cells from their malignant counterparts. Furthermore, we propose that the ability to sense BM appropriately and to form three-dimensional organotypic structures may be the function of a class of "suppressor" genes that are lost as cells become malignant.

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  • Wellings SR, Jensen HM, Marcum RG. An atlas of subgross pathology of the human breast with special reference to possible precancerous lesions. J Natl Cancer Inst. 1975 Aug;55(2):231–273. [PubMed]
  • Dairkee SH, Blayney C, Smith HS, Hackett AJ. Monoclonal antibody that defines human myoepithelium. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7409–7413. [PMC free article] [PubMed]
  • Rudland PS. Stem cells and the development of mammary cancers in experimental rats and in humans. Cancer Metastasis Rev. 1987;6(1):55–83. [PubMed]
  • Hilkens J, Buijs F, Ligtenberg M. Complexity of MAM-6, an epithelial sialomucin associated with carcinomas. Cancer Res. 1989 Feb 15;49(4):786–793. [PubMed]
  • Petersen OW, van Deurs B. Characterization of epithelial membrane antigen expression in human mammary epithelium by ultrastructural immunoperoxidase cytochemistry. J Histochem Cytochem. 1986 Jun;34(6):801–809. [PubMed]
  • Wicha MS, Lowrie G, Kohn E, Bagavandoss P, Mahn T. Extracellular matrix promotes mammary epithelial growth and differentiation in vitro. Proc Natl Acad Sci U S A. 1982 May;79(10):3213–3217. [PMC free article] [PubMed]
  • Lee EY, Lee WH, Kaetzel CS, Parry G, Bissell MJ. Interaction of mouse mammary epithelial cells with collagen substrata: regulation of casein gene expression and secretion. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1419–1423. [PMC free article] [PubMed]
  • Li ML, Aggeler J, Farson DA, Hatier C, Hassell J, Bissell MJ. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. Proc Natl Acad Sci U S A. 1987 Jan;84(1):136–140. [PMC free article] [PubMed]
  • Blum JL, Zeigler ME, Wicha MS. Regulation of rat mammary gene expression by extracellular matrix components. Exp Cell Res. 1987 Dec;173(2):322–340. [PubMed]
  • Barcellos-Hoff MH, Aggeler J, Ram TG, Bissell MJ. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development. 1989 Feb;105(2):223–235. [PMC free article] [PubMed]
  • Streuli CH, Bailey N, Bissell MJ. Control of mammary epithelial differentiation: basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity. J Cell Biol. 1991 Dec;115(5):1383–1395. [PMC free article] [PubMed]
  • Darcy KM, Black JD, Hahm HA, Ip MM. Mammary organoids from immature virgin rats undergo ductal and alveolar morphogenesis when grown within a reconstituted basement membrane. Exp Cell Res. 1991 Sep;196(1):49–65. [PubMed]
  • Stampfer MR, Bartley JC. Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo[a]pyrene. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2394–2398. [PMC free article] [PubMed]
  • Petersen OW, van Deurs B. Growth factor control of myoepithelial-cell differentiation in cultures of human mammary gland. Differentiation. 1988 Dec;39(3):197–215. [PubMed]
  • Band V, Zajchowski D, Kulesa V, Sager R. Human papilloma virus DNAs immortalize normal human mammary epithelial cells and reduce their growth factor requirements. Proc Natl Acad Sci U S A. 1990 Jan;87(1):463–467. [PMC free article] [PubMed]
  • Yaswen P, Smoll A, Peehl DM, Trask DK, Sager R, Stampfer MR. Down-regulation of a calmodulin-related gene during transformation of human mammary epithelial cells. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7360–7364. [PMC free article] [PubMed]
  • Taylor-Papadimitriou J, Stampfer M, Bartek J, Lewis A, Boshell M, Lane EB, Leigh IM. Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: relation to in vivo phenotypes and influence of medium. J Cell Sci. 1989 Nov;94(Pt 3):403–413. [PubMed]
  • Hammond SL, Ham RG, Stampfer MR. Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5435–5439. [PMC free article] [PubMed]
  • Boyd JA, Rinehart CA, Jr, Walton LA, Siegal GP, Kaufman DG. Ultrastructural characterization of two new human endometrial carcinoma cell lines and normal human endometrial epithelial cells cultured on extracellular matrix. In Vitro Cell Dev Biol. 1990 Jul;26(7):701–708. [PubMed]
  • White TE, di Sant'Agnese PA, Miller RK. Human endometrial cells grown on an extracellular matrix form simple columnar epithelia and glands. In Vitro Cell Dev Biol. 1990 Jun;26(6):636–642. [PubMed]
  • Coopman PJ, Bracke ME, Lissitzky JC, De Bruyne GK, Van Roy FM, Foidart JM, Mareel MM. Influence of basement membrane molecules on directional migration of human breast cell lines in vitro. J Cell Sci. 1991 Mar;98(Pt 3):395–401. [PubMed]
  • Satyaswaroop PG, Tabibzadeh SS. Extracellular matrix and the patterns of differentiation of human endometrial carcinomas in vitro and in vivo. Cancer Res. 1991 Oct 15;51(20):5661–5666. [PubMed]
  • Thompson EW, Paik S, Brünner N, Sommers CL, Zugmaier G, Clarke R, Shima TB, Torri J, Donahue S, Lippman ME, et al. Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol. 1992 Mar;150(3):534–544. [PubMed]
  • Petersen OW, van Deurs B. Preservation of defined phenotypic traits in short-term cultured human breast carcinoma derived epithelial cells. Cancer Res. 1987 Feb 1;47(3):856–866. [PubMed]
  • Briand P, Petersen OW, Van Deurs B. A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium. In Vitro Cell Dev Biol. 1987 Mar;23(3):181–188. [PubMed]
  • Soule HD, Maloney TM, Wolman SR, Peterson WD, Jr, Brenz R, McGrath CM, Russo J, Pauley RJ, Jones RF, Brooks SC. Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res. 1990 Sep 15;50(18):6075–6086. [PubMed]
  • Gaffney EV. A cell line (HBL-100) established from human breast milk. Cell Tissue Res. 1982;227(3):563–568. [PubMed]
  • Rønnov-Jessen L, Van Deurs B, Nielsen M, Petersen OW. Identification, paracrine generation, and possible function of human breast carcinoma myofibroblasts in culture. In Vitro Cell Dev Biol. 1992 Apr;28A(4):273–283. [PubMed]
  • Engel LW, Young NA. Human breast carcinoma cells in continuous culture: a review. Cancer Res. 1978 Nov;38(11 Pt 2):4327–4339. [PubMed]
  • Briand P, Lykkesfeldt AE. Long-term cultivation of a human breast cancer cell line, MCF-7, in a chemically defined medium. Effect of estradiol. Anticancer Res. 1986 Jan-Feb;6(1):85–90. [PubMed]
  • Fogh J, Wright WC, Loveless JD. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J Natl Cancer Inst. 1977 Feb;58(2):209–214. [PubMed]
  • Rønnov-Jessen L, Celis JE, Van Deurs B, Petersen OW. A fibroblast-associated antigen: characterization in fibroblasts and immunoreactivity in smooth muscle differentiated stromal cells. J Histochem Cytochem. 1992 Apr;40(4):475–486. [PubMed]
  • Caron de Fromentel C, Nardeux PC, Soussi T, Lavialle C, Estrade S, Carloni G, Chandrasekaran K, Cassingena R. Epithelial HBL-100 cell line derived from milk of an apparently healthy woman harbours SV40 genetic information. Exp Cell Res. 1985 Sep;160(1):83–94. [PubMed]
  • Bissell MJ. The differentiated state of normal and malignant cells or how to define a "normal" cell in culture. Int Rev Cytol. 1981;70:27–100. [PubMed]
  • Blum JL, Zeigler ME, Wicha MS. Regulation of mammary differentiation by the extracellular matrix. Environ Health Perspect. 1989 Mar;80:71–83. [PMC free article] [PubMed]
  • Stampfer M, Hallowes RC, Hackett AJ. Growth of normal human mammary cells in culture. In Vitro. 1980 May;16(5):415–425. [PubMed]
  • Trask DK, Band V, Zajchowski DA, Yaswen P, Suh T, Sager R. Keratins as markers that distinguish normal and tumor-derived mammary epithelial cells. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2319–2323. [PMC free article] [PubMed]
  • Yang J, Balakrishnan A, Hamamoto S, Elias JJ, Rosenau W, Beattie CW, Das Gupta TK, Wellings SR, Nandi S. Human breast epithelial cells in serum-free collagen gel primary culture: growth, morphological, and immunocytochemical analysis. J Cell Physiol. 1987 Nov;133(2):228–255. [PubMed]
  • Foster CS, Smith CA, Dinsdale EA, Monaghan P, Neville AM. Human mammary gland morphogenesis in vitro: the growth and differentiation of normal breast epithelium in collagen gel cultures defined by electron microscopy, monoclonal antibodies, and autoradiography. Dev Biol. 1983 Mar;96(1):197–216. [PubMed]
  • Dulbecco R, Armstrong B, Allen R. Reversion toward an earlier stage of differentiation and loss of polarity during progression of N-methyl-N-nitrosourea-induced rat mammary tumours. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9292–9296. [PMC free article] [PubMed]
  • Kramer RH, Bensch KG, Wong J. Invasion of reconstituted basement membrane matrix by metastatic human tumor cells. Cancer Res. 1986 Apr;46(4 Pt 2):1980–1989. [PubMed]
  • Fridman R, Kibbey MC, Royce LS, Zain M, Sweeney M, Jicha DL, Yannelli JR, Martin GR, Kleinman HK. Enhanced tumor growth of both primary and established human and murine tumor cells in athymic mice after coinjection with Matrigel. J Natl Cancer Inst. 1991 Jun 5;83(11):769–774. [PubMed]
  • Fridman R, Giaccone G, Kanemoto T, Martin GR, Gazdar AF, Mulshine JL. Reconstituted basement membrane (matrigel) and laminin can enhance the tumorigenicity and the drug resistance of small cell lung cancer cell lines. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6698–6702. [PMC free article] [PubMed]
  • Zajchowski DA, Band V, Trask DK, Kling D, Connolly JL, Sager R. Suppression of tumor-forming ability and related traits in MCF-7 human breast cancer cells by fusion with immortal mammary epithelial cells. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2314–2318. [PMC free article] [PubMed]

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