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Role of CEACAM1 isoforms in an in vivo model of mammary morphogenesis: mutational analysis of the cytoplasmic domain of CEACAM1-4S reveals key residues involved in lumen formation

Oncogene volume 26, pages 7637–7646 (2007)Cite this article

Abstract

CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1) is a type I transmembrane glycoprotein expressed in epithelial cells with three or four extracellular domains (ECDs) and either long or short cytoplasmic domain isoforms. We have previously shown that the four extracellular domains, short cytoplasmic domain isoform, CEACAM1-4S, plays an essential role in lumen formation in an in vitro model of mammary morphogenesis. In this study, we transfected MCF-7 cells with either the long or short cytoplasmic domain isoforms of CEACAM1, and grew the cells in humanized mammary mouse fat pads in NOD/SCID mice. In this in vivo model, only the long cytoplasmic domain isoform, CEACAM1-4L, formed glands with lumen. On the basis of other studies that revealed phosphorylation of key Thr and Ser residues in the short cytoplasmic domain, we introduced phosphorylation mimic (for example, Thr or Ser to Asp) or null (Thr or Ser to Ala) mutations into the cytoplasmic domain of CEACAM1-4S and tested them in the in vivo model. Mutation of either Thr or Ser to Asp or the double mutant Thr+Ser to Asp, but not the null mutants, induced gland formation with a central lumen-containing apoptotic cells. Moreover, the phosphorylation mimic mutants of CEACAM1-4S induced downregulation of β1-integrin, overexpression of β2-integrin, inhibited phosphorylation of focal adhesion kinase (pTyr-397) and resulted in myofibroblast differentiation as characterized by expression of vimentin, α-smooth muscle actin and β2-integrin, as well as the production of abundant extracellular matrix.

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References

  • Barnett TR, Drake L, Pickle II W . (1993). Human biliary glycoprotein gene: characterization of a family of novel alternatively spliced RNAs and their expressed proteins. Mol Cell Biol 13: 1273–1282.

    Article  CAS  Google Scholar 

  • Brummer J, Neumaier M, Gopfert C, Wagener C . (1995). Association of pp60c-src with biliary glycoprotein (CD66a), an adhesion molecule of the carcinoembryonic antigen family downregulated in colorectal carcinomas. Oncogene 11: 1649–1655.

    CAS  Google Scholar 

  • Busch C, Hanssen TA, Wagener C, Obrink B . (2002). Down-regulation of CEACAM1 in human prostate cancer: correlation with loss of cell polarity increased proliferation rate, Gleason grade 3 to 4 transition. Hum Pathol 33: 290–298.

    Article  CAS  Google Scholar 

  • Chen CJ, Kirshner J, Sherman MA, Hu W, Nguyen T, Shively JE . (2007). Mutation analysis of the short cytoplasmic domain of the cell-cell adhesion molecule CEACAM1 identifies residues that orchestrate actin binding and lumen formation. J Biol Chem 282: 5749–5760.

    Article  CAS  Google Scholar 

  • Edlund M, Wikstrom K, Toomik R, Ek P, Obrink B . (1998). Characterization of protein kinase C-mediated phosphorylation of the short cytoplasmic domain isoform of C-CAM. FEBS Lett 425: 166–170.

    Article  CAS  Google Scholar 

  • Greenberg RS, Bernstein AM, Benezra M, Gelman IH, Taliana L, Masur SK . (2006). FAK-dependent regulation of myofibroblast differentiation. Faseb J 20: 1006–1008.

    Article  CAS  Google Scholar 

  • Huang J, Hardy JD, Sun Y, Shively JE . (1999). Essential role of biliary glycoprotein (CD66a) in morphogenesis of the human mammary epithelial cell line MCF10F. J Cell Sci 112: 4193–4205.

    CAS  Google Scholar 

  • Huang J, Simpson JF, Glackin C, Riethorf L, Wagener C, Shively JE . (1998). Expression of biliary glycoprotein (CD66a) in normal and malignant breast epithelial cells. Anticancer Res 18: 3203–3212.

    CAS  Google Scholar 

  • Huber M, Izzi L, Grondin P, Houde C, Kunath T, Veillette A et al. (1999). The carboxyl-terminal region of biliary glycoprotein controls its tyrosine phosphorylation and association with protein-tyrosine phosphatases SHP-1 and SHP-2 in epithelial cells. J Biol Chem 274: 335–344.

    Article  CAS  Google Scholar 

  • Kirshner J, Chen CJ, Liu P, Huang J, Shively JE . (2003a). CEACAM1-4S, 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 100: 521–526.

    Article  CAS  Google Scholar 

  • Kirshner J, Schumann D, Shively JE . (2003b). CEACAM1, a cell–cell adhesion molecule, directly associates with annexin II in a three-dimensional model of mammary morphogenesis. J Biol Chem 278: 50338–50345.

    Article  CAS  Google Scholar 

  • Kleinerman DI, Dinney CP, Zhang WW, Lin SH, Van NT, Hsieh JT . (1996a). Suppression of human bladder cancer growth by increased expression of C-CAM1 gene in an orthotopic model. Cancer Res 56: 3431–3435.

    CAS  Google Scholar 

  • Kleinerman DI, Zhang WW, Lin SH, Nguyen TV, von Eschenbach AC, Hsieh JT . (1995b). Application of a tumor suppressor (C-CAM1)-expressing recombinant adenovirus in androgen-independent human prostate cancer therapy: a preclinical study. Cancer Res 55: 2831–2836.

    CAS  Google Scholar 

  • Kunath T, Ordonez-Garcia C, Turbide C, Beauchemin N . (1995). Inhibition of colonic tumor cell growth by biliary glycoprotein. Oncogene 11: 2375–2382.

    CAS  Google Scholar 

  • Kuperwasser C, Chavarria T, Wu M, Magrane G, Gray JW, Carey L et al. (2004). Reconstruction of functionally normal and malignant human breast tissues in mice. Proc Natl Acad Sci USA 101: 4966–4971.

    Article  CAS  Google Scholar 

  • Laurie NA, Comegys MM, Carreiro MP, Brown JF, Flanagan DL, Brilliant KE et al. (2005). Carcinoembryonic antigen-related cell adhesion molecule 1a-4L suppression of rat hepatocellular carcinomas. Cancer Res 65: 11010–11017.

    Article  CAS  Google Scholar 

  • Leers MP, Kolgen W, Bjorklund V, Bergman T, Tribbick G, Persson B et al. (1999). Immunocytochemical detection and mapping of a cytokeratin 18 neo-epitope exposed during early apoptosis. J Pathol 187: 567–572.

    Article  CAS  Google Scholar 

  • Luo W, Tapolsky M, Earley K, Wood CG, Wilson DR, Logothetis CJ et al. (1999). Tumor-suppressive activity of CD66a in prostate cancer. Cancer Gene Ther 6: 313–321.

    Article  CAS  Google Scholar 

  • Luo W, Wood CG, Earley K, Hung MC, Lin SH . (1997). 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 14: 1697–1704.

    Article  CAS  Google Scholar 

  • Mitra SK, Schlaepfer DD . (2006). Integrin-regulated FAK-Src signaling in normal and cancer cells. Curr Opin Cell Biol 18: 516–523.

    Article  CAS  Google Scholar 

  • Neumaier M, Paululat S, Chan A, Matthaes P, Wagener C . (1993). Biliary glycoprotein, a potential human cell adhesion molecule, is down-regulated in colorectal carcinomas. Proc Natl Acad Sci USA 90: 10744–10748.

    Article  CAS  Google Scholar 

  • Nittka S, Gunther J, Ebisch C, Erbersdobler A, Neumaier M . (2004). The human tumor suppressor CEACAM1 modulates apoptosis and is implicated in early colorectal tumorigenesis. Oncogene 23: 9306–9313.

    Article  CAS  Google Scholar 

  • Nollau P, Scheller H, Kona-Horstmann M, Rohde S, Hagenmuller F, Wagener C et al. (1997). Dysregulation of carcinoembryonic antigen group members CGM2, CD66a (biliary glycoprotein), and nonspecific cross-reacting antigen in colorectal carcinomas. Comparative analysis by northern blot and in situ hybridization. Cancer Res 57: 2354–2357.

    CAS  Google Scholar 

  • Obrink B . (1997). CEA adhesion molecules: multifunctional proteins with signal-regulatory properties. Curr Opin Cell Biol 9: 616–626.

    Article  CAS  Google Scholar 

  • Ohwada A, Takahashi H, Nagaoka I, Kira S . (1994). Biliary glycoprotein mRNA expression is increased in primary lung cancer, especially in squamous cell carcinoma. Am J Respir Cell Mol Biol 11: 214–220.

    Article  CAS  Google Scholar 

  • Park CC, Zhang H, Pallavicini M, Gray JW, Baehner F, Park CJ et al. (2006). Beta1 integrin inhibitory antibody induces apoptosis of breast cancer cells, inhibits growth, and distinguishes malignant from normal phenotype in three dimensional cultures and in vivo. Cancer Res 66: 1526–1535.

    Article  CAS  Google Scholar 

  • Peters T, Sindrilaru A, Hinz B, Hinrichs R, Menke A, Al-Azzeh EA et al. (2005). Wound-healing defect of CD18 (−/−) mice due to a decrease in TGF-beta1 and myofibroblast differentiation. Embo J 24: 3400–3410.

    Article  CAS  Google Scholar 

  • Powell DW, Adegboyega PA, Di Mari JF, Mifflin RC . (2005). Epithelial cells and their neighbors I. Role of intestinal myofibroblasts in development, repair, and cancer. Am J Physiol Gastrointest Liver Physiol 289: G2–G7.

    Article  CAS  Google Scholar 

  • Powell DW, Mifflin RC, Valentich JD, Crowe SE, Saada JI, West AB . (1999). Myofibroblasts. II. Intestinal subepithelial myofibroblasts. Am J Physiol 277: C1–C9.

    Article  CAS  Google Scholar 

  • Schumann D, Chen CJ, Kaplan B, Shively JE . (2001). Carcinoembryonic antigen cell adhesion molecule 1 directly associates with cytoskeleton proteins actin and tropomyosin. J Biol Chem 276: 47421–47433.

    Article  CAS  Google Scholar 

  • Shi S-R, Cote RJ, Taylor CR . (2001). Antigen retrieval techniques: current perspectives. J Histochem Cytochem 49: 931–937.

    Article  CAS  Google Scholar 

  • Svenberg T, Hammarstrom S, Zeromski J . (1979). Immunofluorescence studies on the occurrence and localization of the CEA-related biliary glycoprotein I (BGP I) in normal human gastrointestinal tissues. Clin Exp Immunol 36: 436–441.

    CAS  PubMed Central  Google Scholar 

  • Tanaka K, Hinoda Y, Takahashi H, Sakamoto H, Nakajima Y, Imai K . (1997). Decreased expression of biliary glycoprotein in hepatocellular carcinomas. Int J Cancer 74: 15–19.

    Article  CAS  Google Scholar 

  • Tomasek JJ, Gabbiani G, Hinz B, Chaponnier C, Brown RA . (2002). Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat Rev Mol Cell Biol 3: 349–363.

    Article  CAS  Google Scholar 

  • Wagener C, Clark BR, Rickard KJ, Shively JE . (1983). Monoclonal antibodies for carcinoembryonic antigen and related antigens as a model system: a systematic approach for the determination of epitope specificities of monoclonal antibodies. J Immunol 130: 2302–2307.

    CAS  Google Scholar 

Download references

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  1. Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, CA, USA

    S Yokoyama, C-J Chen, T Nguyen & J E Shively

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  1. S Yokoyama
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  2. C-J Chen
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  3. T Nguyen
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  4. J E Shively
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Correspondence to J E Shively.

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Yokoyama, S., Chen, CJ., Nguyen, T. et al. Role of CEACAM1 isoforms in an in vivo model of mammary morphogenesis: mutational analysis of the cytoplasmic domain of CEACAM1-4S reveals key residues involved in lumen formation. Oncogene 26, 7637–7646 (2007). https://doi.org/10.1038/sj.onc.1210577

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