Abstract
CEA cell adhesion molecule-1 (CEACAM1) is a cell–cell adhesion molecule that, paradoxically, is expressed in an apical location in normal breast epithelium. Strong lumenal membrane staining is observed in 100% of normal glands (11/11), low in atypical hyperplasia (2/6), high in cribiform ductal carcinoma in situ (DCIS) (8/8), but low in other types of DCIS (2/15). Although most invasive ductal carcinomas express CEACAM1 (21/26), the staining pattern tends to be weak and cytoplasmic in tumours with minimal lumena formation (grades 2–3), while there is membrane staining in well-differentiated tumours (grade 1). The 'normal' breast epithelial line MCF10F forms acini with lumena in Matrigel with apical membrane expression of CEACAM1. MCF7 cells that do not express CEACAM1 and fail to form lumena in Matrigel, revert to a lumen forming phenotype when transfected with the CEACAM1-4S but not the -4L isoform. CEACAM1 directly associates with and down-regulates the expression of β1-integrin. Immuno-electron microscopy reveals numerous vesicles coated with CEACAM1 within the lumena, and as predicted by this finding, CEACAM1 is found in the lipid fraction of breast milk. Thus, CEACAM1 is a critical molecule in mammary morphogenesis and may play a role in the absorption of the lipid vesicles of milk in the infant intestinal tract.
Keywords: Integrin, MCF7 Cell, Cell Adhesion Molecule, Normal Breast, Invasive Ductal Carcinoma
References
- Barcellos-Hoff MH, Aggeler J, Ram TG, Bissell MJ. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development. 1989;105:223–235. doi: 10.1242/dev.105.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barnett TR, Drake L, Pickle WI. Human biliary glycoprotein gene: Characterization of a family of novel alternatively spliced RNAs and their expressed proteins. Mol Cell Biol. 1993;13:1273–1282. doi: 10.1128/mcb.13.2.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barnett TR, Kretschmer A, Austen DA, Goebel SJ, Hart JT, Elting JJ, Kamarck ME. Carcinoembryonic antigens: Alternative splicing accounts for the multiple mRNAs that code for novel members of the carcinoembryonic antigen family. J Cell Biol. 1989;108:267–276. doi: 10.1083/jcb.108.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bergstraesser LM, Weitzman SA. Culture of normal and malignant primary human mammary epithelial cells in a physiological manner. simulates in vivo growth patterns and allows discrimination of cell type. Cancer Res. 1993;53:2644–2654. [PubMed] [Google Scholar]
- Bissell MJ, Radisky DC, Rizki A, Weaver VM, Petersen OW. The organizing principle: Microenvironmental influences in the normal and malignant breast. Differentiation. 2002;70:537–546. doi: 10.1046/j.1432-0436.2002.700907.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bos MP, Kuroki M, Krop-Watorek A, Hogan D, Belland RJ. CD66 receptor specificity exhibited by neisserial Opa variants is controlled by protein determinants in CD66 N-domains. Proc Natl Acad Sci USA. 1998;95:9584–9589. doi: 10.1073/pnas.95.16.9584. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brinkmann V, Foroutan H, Sachs M, Weidner KM, Birchmeier W. Hepatocyte growth factor/scatter factor induces a variety of tissue-specific morphogenic programs in epithelial cells. J Cell Biol. 1995;131:1573–1586. doi: 10.1083/jcb.131.6.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brummer J, Ebrahimnejad A, Flayeh R, Schumacher U, Loning T, Bamberger AM, Wagener C. Cis interaction of the cell adhesion molecule CEACAM1with integrin beta(3) AmJ Pathol. 2001;159:537–546. doi: 10.1016/s0002-9440(10)61725-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buescher ES. Anti-inflammatory characteristics of human milk: How, where, why? Adv Exp Med Biol. 2001;501:207–222. doi: 10.1007/978-1-4615-1371-1_27. [DOI] [PubMed] [Google Scholar]
- Chen T, Bolland S, Chen I, Parker J, Pantelic M, Grunert F, Zimmermann W. The CGM1a (CEACAM3/CD66d)-mediated phagocytic pathway of Neisseria gonorrhoeae expressing opacity proteins is also the pathway to cell death. J Biol Chem. 2001;276:17413–17419. doi: 10.1074/jbc.M010609200. [DOI] [PubMed] [Google Scholar]
- Cross PC, Mercer KL. Cell and Tissue Ultrastructure. New York: W.H. Freeman and Company; 1993. [Google Scholar]
- Drzeniek Z, Lamerz R, Fenger U, Wagener C, Haubeck HD. Identification of membrane antigens in granulocytes and colonic carcinoma cells by a monoclonal antibody specific for biliary glycoprotein, a member of the carcinoembryonic antigen family. Cancer Lett. 1991;56:173–179. doi: 10.1016/0304-3835(91)90093-w. [DOI] [PubMed] [Google Scholar]
- Dvesksler GS, Dieffenbach CW, Cardellichio CB, McCuaig K, Pensiero MN, Jiang G-S, Beauchemin N, Holmes KV. Several members of the mouse carcinoembryonic antigen-related glycoprotein family are functional receptors for the coronavirus mouse hepatitis virus A-59. J Virol. 1993;67:1–8. doi: 10.1128/jvi.67.1.1-8.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Frangsmyr L, Baranov V, Prall F, Yeung MM-W, Wagener C, Hammarstrom S. Cell and region specific expression of biliary glycoprotein and its messenger RNAin normal human colonic mucosa. Cancer Res. 1995;55:2963–2967. [PubMed] [Google Scholar]
- Gomm JJ, Coope RC, Browne PJ, Coombes RC. Separated human breast epithelial and myoepithelial cells have different growth factor requirements in vitro but can reconstitute normal breast lobuloalveolar structure. J Cell Physiol. 1997;171:11–19. doi: 10.1002/(SICI)1097-4652(199704)171:1<11::AID-JCP2>3.0.CO;2-K. [DOI] [PubMed] [Google Scholar]
- Gray-Owen SD, Lorenzen DR, Haude A, Meyer TF, Dehio C. Differential Opa specificities for CD66 receptors influence tissue inter-actions and cellular response to Neisseria gonorrhoeae. Mol Microbiol. 1997;26:971–980. doi: 10.1046/j.1365-2958.1997.6342006.x. [DOI] [PubMed] [Google Scholar]
- Hakansson A, Zhivotovsky B, Orrenius S, Sabharwal H, Svanborg C. Apoptosis induced by a human milk protein. Proc Natl Acad Sci USA. 1995;92:8064–8068. doi: 10.1073/pnas.92.17.8064. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hinoda Y, Neumaier M, Hefta SA, Drzeniek Z, Wagener C, Shively L, Hefta LJF, Shively JE, Paxton RJ. Molecular cloning of a cDNA coding biliary glycoprotein I: Primary structure of a glycoprotein immunologically crossreactive with carcinoembryonic antigen. Proc Natl Acad Sci USA. 1988;85:6959–6963. doi: 10.1073/pnas.85.18.6959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirai Y, Lochter A, Galosy S, Koshida S, Niwa S, Bissell MJ. Epimorphin functions as a key morphoregulator for mammary epithelial cells. J Cell Biol. 1998;140:159–169. doi: 10.1083/jcb.140.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsieh J.-T., Luo W, Song W, Wang Y, Kleinerman DI, Van NT, Lin S-H. Tumor suppressive role of an androgen-regulated epithelial cell adhesion molecule (C-CAM) in prostate carcinoma cell revealed by sense and anti-sense approaches. Cancer Res. 1995;55:190–197. [PubMed] [Google Scholar]
- Huang J, Hardy JD, Sun Y, Shively JE. Essential role of biliary glycoprotein (CD66a) in morphogenesis of the human mammary epithelial cell line MCF10F. J Cell Sci. 1999;112:4193–4205. doi: 10.1242/jcs.112.23.4193. [DOI] [PubMed] [Google Scholar]
- Huang J, Simpson JF, Glackin C, Riethorf L, Wagener C, Shively JE. Expression of biliary glycoprotein (CD66a) in normal and malignant breast epithelial cells. Anticancer Res. 1998;18:3203–3212. [PubMed] [Google Scholar]
- Hudson DL, O'Hare M, Watt FM, Masters JR. Proliferative heterogeneity in the human prostate: Evidence for epithelial stem cells. Lab Invest. 2000;80:1243–1250. doi: 10.1038/labinvest.3780132. [DOI] [PubMed] [Google Scholar]
- Kirshner J, Chen CJ, Liu P, Huang J, Shively JE. CEACAM1–1S, a cell–cell adhesion molecule, mediates apoptosis and reverts mammary carcinoma cells to a normal morphogenic phenotype in a 3D culture. Proc Natl Acad Sci USA. 2003;100:521–526. doi: 10.1073/pnas.232711199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kleinerman DI, Dinney CP, Zhang WW, Lin SH, Van NT, Hsieh JT. Suppression of human bladder cancer growth by increased expression of C-CAM1 gene in an orthotopic model. Cancer Res. 1996;56:3431–3435. [PubMed] [Google Scholar]
- Kleinerman DI, Troncoso P, Lin S-H, Pister LL, Sherwood ER, Brooks T, von Eschenbach AC, Hseih J-T. Consistent expression of an epithelial cell adhesion molecule (C-CAM) during prostate development and loss of expression in prostate cancer: Implication as a tumor suppressor. Cancer Res. 1995;55:1215–1220. [PubMed] [Google Scholar]
- Kleinerman DI, Zhang W-W, Lin S-H, Van NT, von Eschenbach AC, Hseih J-T. Application of a tumor suppressor (C-CAM1)-expressing recombinant adenovirus in androgen-independent human prostate cancer therapy: A preclinical study. Cancer Res. 1995;55:2831–2836. [PubMed] [Google Scholar]
- Klinowska TC, Soriano JV, Edwards GM, Oliver JM, Valentijn AJ, Montesano R, Streuli CH. Laminin and beta1 integrins are crucial for normal mammary gland development in the mouse. Dev Biol. 1999;215:13–32. doi: 10.1006/dbio.1999.9435. [DOI] [PubMed] [Google Scholar]
- Li M, Liu X, Robinson G, Bar-Peled U, Wagner KU, Young WS, Hennighausen L, Furth PA. Mammary-derived signals activate programmed cell death during the first stage of mammary gland involution. Proc Natl Acad Sci USA. 1997;94:3425–3430. doi: 10.1073/pnas.94.7.3425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li S, Shipley GD. Expression of multiple species of basic fibroblast growth factor mRNA and protein in normal and tumor-derived mammary epithelial cells in culture. Cell Growth Differ. 1991;2:195–202. [PubMed] [Google Scholar]
- Lin S-H, Guidotti G. Cloning and expression of a cDNA coding for a rat liver plasma membrane ecto-ATPase. J Biol Chem. 1989;264:14408–14414. [PubMed] [Google Scholar]
- Lochter A, Navre M, Werb Z, Bissell MJ. Alpha1 and alpha2 integrins mediate invasive activity of mouse mammary carcinoma cells through regulation of stromelysin-1 expression. Mol Biol Cell. 1999;10:271–282. doi: 10.1091/mbc.10.2.271. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luo W, Wood CG, Earley K, Hung M-C, Lin SH. Suppression of tumorigenicity of breast cancer cells by an epithelial cell adhesion molecule (C-CAM1): The adhesion and growth suppression are mediated by different domains. Oncogene. 1997;14:1697–16704. doi: 10.1038/sj.onc.1200999. [DOI] [PubMed] [Google Scholar]
- Mather IH, Jack LJ, Madara PJ, Johnson VG. The distribution of MUC1, an apical membrane glycoprotein, in mammary epithelial cells at the resolution of the electron microscope: Implications for the mechanism of milk secretion. Cell Tissue Res. 2001;304:91–101. doi: 10.1007/s004410100351. [DOI] [PubMed] [Google Scholar]
- Mather IH, Keenan TW. Origin and secretion of milk lipids. J Mammary Gland Biol Neoplasia. 1998;3:259–273. doi: 10.1023/a:1018711410270. [DOI] [PubMed] [Google Scholar]
- Muenzner P, Naumann M, Meyer TF, Gray-Owen SD. Pathogenic Neisseria trigger expression of their carcinoembryonic antigen-related cellular adhesion molecule 1 (CEACAM1; previously CD66a) receptor on primary endothelial cells by activating the immediate early response transcription factor, nuclear factor-kappaB. J Biol Chem. 2001;276:24331–24340. doi: 10.1074/jbc.M006883200. [DOI] [PubMed] [Google Scholar]
- Muschler J, Lochter A, Roskelley CD, Yurchenco P, Bissell MJ. Division of labor among the alpha6beta4 integrin, beta1 integrins, and an E3 laminin receptor to signal morphogenesis and beta-casein expression in mammary epithelial cells. Mol Biol Cell. 1999;10:2817–2828. doi: 10.1091/mbc.10.9.2817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muthuswamy SK, Li D, Lelievre S, Bissell MJ, Brugge JS. ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini. Nat Cell Biol. 2001;3:785–792. doi: 10.1038/ncb0901-785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Najjar SM, Accili D, Philippe N, Jernberg J, Margolis R, Taylor SI. pp120/ecto-ATPase, an endogenase substrate of the insulin receptor tyrosine kinase, is expressed as two variably spliced isoforms. J Biol Chem. 1993;268:1201–1206. [PubMed] [Google Scholar]
- Najjar SM, Philippe N, Suzuki Y, Ignacio GA, Formisano P, Accili D, Taylor SI. Insulin-stimulated phosphorylation of recombinant pp120/HA4, an endogenous substrate of the insulin receptor tyrosine kinase. Biochemistry. 1995;34:9341–9349. doi: 10.1021/bi00029a009. [DOI] [PubMed] [Google Scholar]
- Neumaier M, Paululat S, Chan A, Matthaes P, Wagener C. Biliary glycoprotein, a potential human cell adhesion molecule, is down-regulated in colorectal carcinomas. Proc Natl Acad Sci USA. 1993;90:10744–10748. doi: 10.1073/pnas.90.22.10744. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ocklind C, Obrink B. Intercellular adhesion of rat hepatocytes. Identification of a cell surface glycoprotein involved in the initial adhesion process. J Biol Chem. 1982;257:6788–6795. [PubMed] [Google Scholar]
- Oshima K, Aoki N, Kato T, Kitajima K, Matsuda T. Secretion of a peripheral membrane protein, MFG-E8, as a complex with membrane vesicles. Eur J Biochem. 2002;269:1209–1218. doi: 10.1046/j.1432-1033.2002.02758.x. [DOI] [PubMed] [Google Scholar]
- Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ. Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA. 1992;89:9064–9068. doi: 10.1073/pnas.89.19.9064. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poy MN, Ruch RJ, Fernstrom MA, Okabayashi Y, Najjar SM. Shc and CEACAM1interact to regulate the mitogenic action of insulin. J Biol Chem. 2002;277:1076–1084. doi: 10.1074/jbc.M108415200. [DOI] [PubMed] [Google Scholar]
- Poy MN, Yang Y, Rezaei K, Fernstrom MA, Lee AD, Kido Y, Erickson SK, Najjar SM. CEACAM1 regulates insulin clearance in liver. Nat Genet. 2002;30:270–276. doi: 10.1038/ng840. [DOI] [PubMed] [Google Scholar]
- Riethdorf L, Lisboa BW, Henkel U, Naumann M, Wagener C, Loning T. Differential expression of CD66a (BGP), a cell adhesion molecule of the carcinoembryonic antigen family, in benign, premalignant, and malignant lesions of the human mammary gland. J Histochem Cytochem. 1997;45:957–963. doi: 10.1177/002215549704500705. [DOI] [PubMed] [Google Scholar]
- Rudloff S, Stefan C, Pohlentz G, Kunz C. Detection of ligands for selectins in the oligosaccharide fraction of human milk. Eur J Nutr. 2002;41:85–92. doi: 10.1007/s003940200012. [DOI] [PubMed] [Google Scholar]
- Sippel CJ, Suchy FJ, Ananthanarayanan M, Perlmutter DH. The rat liver ecto-ATPase is also a canalicular bile acid transport protein. J Biol Chem. 1993;268:2083–2091. [PubMed] [Google Scholar]
- Soriano JV, Pepper MS, Nakamura T, Orci L, Montesano R. Hepatocyte growth factor stimulates extensive development of branching duct-like structures by cloned mammary gland epithelial cells. J Cell Sci. 1995;108:413–430. doi: 10.1242/jcs.108.2.413. [DOI] [PubMed] [Google Scholar]
- Svenberg T. Carcinoembryonic antigen-like substances of human bile. Isolation and partial characterization. Int J Cancer. 1976;17:588–596. doi: 10.1002/ijc.2910170506. [DOI] [PubMed] [Google Scholar]
- Svenberg T, Hammarstrom S, Zeromski J. Immunofluorescence studies on the occurrence and localization of the CEA-related biliary glycoprotein I (BGP I) in normal human gastrointestinal tissues. Clin Exp Immunol. 1979;36:436–441. [PMC free article] [PubMed] [Google Scholar]
- Takahashi H, Okai Y, Paxton RJ, Hefta LJ, Shively JE. Differential regulation of carcinoembryonic antigen and biliary glycoprotein by gamma-interferon. Cancer Res. 1993;53:1612–1619. [PubMed] [Google Scholar]
- Virji M, Evans D, Griffith J, Hill D, Serino L, Hadfield A, Watt SM. Carcinoembryonic antigens are targeted by diverse strains of typable and non-typable Haemophilus influenzae. Mol Microbiol. 2000;36:784–795. doi: 10.1046/j.1365-2958.2000.01885.x. [DOI] [PubMed] [Google Scholar]
- Virji M, Makepeace K, Ferguson DJ, Watt SM. Carcinoembryonic antigens (CD66) on epithelial cells and neutrophils are receptors for Opa proteins of pathogenic neisseriae. Mol Microbiol. 1996;22:941–950. doi: 10.1046/j.1365-2958.1996.01551.x. [DOI] [PubMed] [Google Scholar]
- Weaver VM, Petersen OW, Wang F, Larabell CA, Briand P, Damsky C, Bissell MJ. Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J Cell Biol. 1997;137:231–245. doi: 10.1083/jcb.137.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Xyni K, Rizos D, Giannaki G, Sarandakou A, Phocas I, Creatsas G. Soluble form of ICAM-1, VCAM-1, E-and L-selectin in human milk. Mediators Inflamm. 2000;9:133–140. doi: 10.1080/09629350020003858. [DOI] [PMC free article] [PubMed] [Google Scholar]