Skip to main content
British Journal of Cancer logoLink to British Journal of Cancer
. 1991 Oct;64(4):710–714. doi: 10.1038/bjc.1991.386

Establishment and characterisation of human carcinoembryonic antigen transgenic mice.

T Hasegawa 1, K Isobe 1, Y Tsuchiya 1, S Oikawa 1, H Nakazato 1, H Ikezawa 1, I Nakashima 1, K Shimokata 1
PMCID: PMC1977681  PMID: 1911219

Abstract

We have produced human CEA transgenic mice which were found to express CEA mRNA in all tissues. By immunoblot analysis using anti-CEA polyclonal antibody, we also detected CEA protein in all tissues. However, the molecular size of CEA in the brain was different from that in other tissues, although the mRNA size was same and no deletion nor rearrangement was detected at the DNA level. Immunohistochemical analysis of the lung and the colon showed that the expression sites were the bronchial epithelial cells of the lung and the columnar epithelial cells of the colon. Interestingly, the expression of CEA protein in the transgenic mice was polarised to the luminal side of epithelial cells similar to the normal CEA expression in human tissues. We also detected cell surface expression of human CEA on thymocytes and spleen cells and CEA expression was greatly reduced by the phosphatidylinositol-specific phospholipase C (PI-PLC) treatment.

Full text

PDF
710

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beauchemin N., Benchimol S., Cournoyer D., Fuks A., Stanners C. P. Isolation and characterization of full-length functional cDNA clones for human carcinoembryonic antigen. Mol Cell Biol. 1987 Sep;7(9):3221–3230. doi: 10.1128/mcb.7.9.3221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beauchemin N., Turbide C., Afar D., Bell J., Raymond M., Stanners C. P., Fuks A. A mouse analogue of the human carcinoembryonic antigen. Cancer Res. 1989 Apr 15;49(8):2017–2021. [PubMed] [Google Scholar]
  3. Benchimol S., Fuks A., Jothy S., Beauchemin N., Shirota K., Stanners C. P. Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule. Cell. 1989 Apr 21;57(2):327–334. doi: 10.1016/0092-8674(89)90970-7. [DOI] [PubMed] [Google Scholar]
  4. Boucher D., Cournoyer D., Stanners C. P., Fuks A. Studies on the control of gene expression of the carcinoembryonic antigen family in human tissue. Cancer Res. 1989 Feb 15;49(4):847–852. [PubMed] [Google Scholar]
  5. Buchegger F., Schreyer M., Carrel S., Mach J. P. Monoclonal antibodies identify a CEA crossreacting antigen of 95 kD (NCA-95) distinct in antigenicity and tissue distribution from the previously described NCA of 55 kD. Int J Cancer. 1984 May 15;33(5):643–649. doi: 10.1002/ijc.2910330515. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Egan M. L., Pritchard D. G., Todd C. W., Go V. L. Isolation and immunochemical and chemical characterization of carcinoembryonic antigen-like substances in colon lavages of healthy individuals. Cancer Res. 1977 Aug;37(8 Pt 1):2638–2643. [PubMed] [Google Scholar]
  8. Fritsche R., Mach J. P. Isolation and characterization of carcinoembryonic antigen (CEA) extracted from normal human colon mucosa. Immunochemistry. 1977 Feb;14(2):119–127. doi: 10.1016/0019-2791(77)90290-7. [DOI] [PubMed] [Google Scholar]
  9. GOLD P., FREEDMAN S. O. DEMONSTRATION OF TUMOR-SPECIFIC ANTIGENS IN HUMAN COLONIC CARCINOMATA BY IMMUNOLOGICAL TOLERANCE AND ABSORPTION TECHNIQUES. J Exp Med. 1965 Mar 1;121:439–462. doi: 10.1084/jem.121.3.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gold P., Freedman S. O. Specific carcinoembryonic antigens of the human digestive system. J Exp Med. 1965 Sep 1;122(3):467–481. doi: 10.1084/jem.122.3.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hefta L. J., Schrewe H., Thompson J. A., Oikawa S., Nakazato H., Shively J. E. Expression of complementary DNA and genomic clones for carcinoembryonic antigen and nonspecific cross-reacting antigen in Chinese hamster ovary and mouse fibroblast cells and characterization of the membrane-expressed products. Cancer Res. 1990 Apr 15;50(8):2397–2403. [PubMed] [Google Scholar]
  12. Hefta S. A., Hefta L. J., Lee T. D., Paxton R. J., Shively J. E. Carcinoembryonic antigen is anchored to membranes by covalent attachment to a glycosylphosphatidylinositol moiety: identification of the ethanolamine linkage site. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4648–4652. doi: 10.1073/pnas.85.13.4648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ikezawa H., Nakabayashi T., Suzuki K., Nakajima M., Taguchi T., Taguchi R. Complete purification of phosphatidylinositol-specific phospholipase C from a strain of Bacillus thuringiensis. J Biochem. 1983 Jun;93(6):1717–1719. doi: 10.1093/oxfordjournals.jbchem.a134315. [DOI] [PubMed] [Google Scholar]
  14. Kamarck M. E., Elting J. J., Hart J. T., Goebel S. J., Rae P. M., Nothdurft M. A., Nedwin J. J., Barnett T. R. Carcinoembryonic antigen family: expression in a mouse L-cell transfectant and characterization of a partial cDNA in bacteriophage lambda gt11. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5350–5354. doi: 10.1073/pnas.84.15.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Low M. G., Kincade P. W. Phosphatidylinositol is the membrane-anchoring domain of the Thy-1 glycoprotein. Nature. 1985 Nov 7;318(6041):62–64. doi: 10.1038/318062a0. [DOI] [PubMed] [Google Scholar]
  16. Mach J. P., Pusztaszeri G. Carcinoembryonic antigen (CEA): demonstration of a partial identity between CEA and a normal glycoprotein. Immunochemistry. 1972 Oct;9(10):1031–1034. doi: 10.1016/0019-2791(72)90113-9. [DOI] [PubMed] [Google Scholar]
  17. Nagura H., Koshikawa T., Fukuda Y., Asai J. Hepatic vascular endothelial cells heterogenously express surface antigens associated with monocytes, macrophages and T lymphocytes. Virchows Arch A Pathol Anat Histopathol. 1986;409(4):407–416. doi: 10.1007/BF00705413. [DOI] [PubMed] [Google Scholar]
  18. Neumaier M., Zimmermann W., Shively L., Hinoda Y., Riggs A. D., Shively J. E. Characterization of a cDNA clone for the nonspecific cross-reacting antigen (NCA) and a comparison of NCA and carcinoembryonic antigen. J Biol Chem. 1988 Mar 5;263(7):3202–3207. [PubMed] [Google Scholar]
  19. O'Hare K., Benoist C., Breathnach R. Transformation of mouse fibroblasts to methotrexate resistance by a recombinant plasmid expressing a prokaryotic dihydrofolate reductase. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1527–1531. doi: 10.1073/pnas.78.3.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oikawa S., Imajo S., Noguchi T., Kosaki G., Nakazato H. The carcinoembryonic antigen (CEA) contains multiple immunoglobulin-like domains. Biochem Biophys Res Commun. 1987 Apr 29;144(2):634–642. doi: 10.1016/s0006-291x(87)80013-x. [DOI] [PubMed] [Google Scholar]
  21. Oikawa S., Inuzuka C., Kuroki M., Matsuoka Y., Kosaki G., Nakazato H. Cell adhesion activity of non-specific cross-reacting antigen (NCA) and carcinoembryonic antigen (CEA) expressed on CHO cell surface: homophilic and heterophilic adhesion. Biochem Biophys Res Commun. 1989 Oct 16;164(1):39–45. doi: 10.1016/0006-291x(89)91679-3. [DOI] [PubMed] [Google Scholar]
  22. Oikawa S., Kosaki G., Nakazato H. Molecular cloning of a gene for a member of carcinoembryonic antigen (CEA) gene family; signal peptide and N-terminal domain sequences of nonspecific crossreacting antigen (NCA). Biochem Biophys Res Commun. 1987 Jul 31;146(2):464–469. doi: 10.1016/0006-291x(87)90552-3. [DOI] [PubMed] [Google Scholar]
  23. Oikawa S., Nakazato H., Kosaki G. Primary structure of human carcinoembryonic antigen (CEA) deduced from cDNA sequence. Biochem Biophys Res Commun. 1987 Jan 30;142(2):511–518. doi: 10.1016/0006-291x(87)90304-4. [DOI] [PubMed] [Google Scholar]
  24. Paxton R. J., Mooser G., Pande H., Lee T. D., Shively J. E. Sequence analysis of carcinoembryonic antigen: identification of glycosylation sites and homology with the immunoglobulin supergene family. Proc Natl Acad Sci U S A. 1987 Feb;84(4):920–924. doi: 10.1073/pnas.84.4.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sato C., Miyaki M., Oikawa S., Nakazato H., Kosaki G. Differential expression of carcinoembryonic antigen and nonspecific crossreacting antigen genes in human colon adenocarcinomas and normal colon mucosa. Jpn J Cancer Res. 1988 Apr;79(4):433–437. doi: 10.1111/j.1349-7006.1988.tb01610.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takami N., Misumi Y., Kuroki M., Matsuoka Y., Ikehara Y. Evidence for carboxyl-terminal processing and glycolipid-anchoring of human carcinoembryonic antigen. J Biol Chem. 1988 Sep 5;263(25):12716–12720. [PubMed] [Google Scholar]
  27. Tate H. Plasma CEA in the post-surgical monitoring of colorectal carcinoma. Br J Cancer. 1982 Sep;46(3):323–330. doi: 10.1038/bjc.1982.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tawaragi Y., Oikawa S., Matsuoka Y., Kosaki G., Nakazato H. Primary structure of nonspecific crossreacting antigen (NCA), a member of carcinoembryonic antigen (CEA) gene family, deduced from cDNA sequence. Biochem Biophys Res Commun. 1988 Jan 15;150(1):89–96. doi: 10.1016/0006-291x(88)90490-1. [DOI] [PubMed] [Google Scholar]
  29. Yamamoto M., Shimokata K., Nagura H. An immunohistochemical study on phenotypic heterogeneity of human pulmonary vascular endothelial cells. Virchows Arch A Pathol Anat Histopathol. 1988;412(5):479–486. doi: 10.1007/BF00750582. [DOI] [PubMed] [Google Scholar]
  30. Zimmermann W., Ortlieb B., Friedrich R., von Kleist S. Isolation and characterization of cDNA clones encoding the human carcinoembryonic antigen reveal a highly conserved repeating structure. Proc Natl Acad Sci U S A. 1987 May;84(9):2960–2964. doi: 10.1073/pnas.84.9.2960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. von Kleist S., Chavanel G., Burtin P. Identification of an antigen from normal human tissue that crossreacts with the carcinoembryonic antigen. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2492–2494. doi: 10.1073/pnas.69.9.2492. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES