A Polymerase-chain-reaction Assay for the Specific Identification of Transcripts Encoded by Individual Carcinoembryonic Antigen (CEA)-gene-family Members

Thompson, John A.; Mössinger, S.; Reichhardt, V.; Engels, U.; Beauchemin, N.; Komoss, F.; Kleist, Sabine von; Zimmermann, Wolfgang (9. September 1993): A Polymerase-chain-reaction Assay for the Specific Identification of Transcripts Encoded by Individual Carcinoembryonic Antigen (CEA)-gene-family Members. In: International Journal of Cancer, Vol. 55, No. 2: pp. 311-319

Abstract

Carcinoembryonic antigen (CEA) is a tumor marker that belongs to a family of closely related molecules with variable expression patterns. We have developed sets of oligonucleotide primers for the specific amplification of transcripts from individual CEA-family members using the reverse transcriptase/ polymerase chain reaction (RT/PCR). Specific primer sets were designed for CEA, non-specific cross-reacting antigen (NCA), biliary glycoprotein (BGP), carcinoembryonic antigen gene-family members 1, 6 and 7 (CGMI, CGM6 and CGM7), and one set for all pregnancy-specific glycoprotein (PSG) transcripts. Primers were first tested for their specificity against individual cDNA clones and product-hybridization with internal, transcript-specific oligonucleotides. Total RNA from 12 brain and 63 gynecological tumors were then tested for expression of CEA-related transcripts. None were found in tumors located in the brain, including various mesenchymal and neuro-epithelial tumors. CEA and NCA transcripts were, however, present in an adenocarcinoma located in the nasal sinuses. In ovarian mucinous adenocarcinomas, we always found co-expression of CEA and NCA transcripts, and occasionally BGP mRNA. CEA-related transcripts were also found in some serous, endometrioid and clear-cell ovarian carcinomas. CEA, NCA and BGP transcripts were present in endometrial carcinomas of the uterus and cervical carcinomas, whereas uterine leiomyomas were completely negative. No transcripts were found from CGM 1, CGM6, CGM7 or from PSG genes in any of the tumors tested. The PCR data were compared with immunohistochemical investigations of ovarian tumors at the protein level using CEA (26/3/13)-, NCA-50/90 (9A6FR) and NCA-95 (80H3)-specific monoclonal antibodies.

• Item Type: Journal article
• Faculties: Medicine
• Subjects: 600 Technology > 610 Medicine and health
• ISSN: 1097-0215

References

Ang, L. C., Tayler, A. R., Bergin, D. and Kaufmann, J. C. E., An immunohistochemical study of papillary tumors in the central nervous system. Cancer , 65, 2712-2719 (1990). Links Berling, B., Kolbinger, F., Grunert, F., Thompson, J. A., Brombacher, F., Buchegger, F., von Kleist, S. and Zimmermann, W., Cloning of a carcinoembryonic-antigen-family member expressed in leukocytes of chronic-myeloid-leukaemia patients and bone marrow. Cancer Res. , 50, 6534-6539 (1990). Links Bernards, A., Rubin, C. M., Westbrook, C. A., Paskind, M. and Baltimore, D., The first intron in the human c-abl gene is at least 200 kilobases long and is a target for translocations in chronic myelogenous leukemia. Mol. cell Biol. , 7, 3231-3236 (1987). Links Chi, K., Jessup, J. M. and Frazier, M. L., Predominant expression of mRNA coding for non-specific cross-reacting antigen in colorectal carcinomas. Tumor Biol. , 12, 298-308 (1991). Links Chomczynski, P. and Qasba, P. K., Alkaline transfer of DNA to plastic membrane. Biochem. biophys. Res. Commun. , 122, 340-344 (1984). Links Chomczynski, P. and Sacchi, N., Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. , 162, 156-159 (1987). Links Daniel, S., Nagel, G., Johnson, J., Lobo, F. M., Hirn, M., Jantscheff, P., Kuroki, M., von Kleist, S. and Grunert, F., Determination of the specifities of monoclonal antibodies recognizing members of the CEA family using a panel of transfectants. Int. J. Cancer , 55, 303-310 (1993). Links Fainstein, E., Einat, M., Gokkel, E., Marcelle, C, Croce, C. M., Gale, R. P. and Canaani, E., Nucleotide sequence analysis of human abl and bcr-abl cDNAs. Oncogene , 4, 1477-1481 (1989). Links Hinoda, Y., Imai, K., Nakagawa, N., Ibayashi, Y., Nakano, T., Paxton, R. J., Shively, J. E. and Yachi, A., Transcription of biliary glycoprotein I in malignant and non-malignant human liver tissues. Int. J. Cancer , 45, 875-878 (1990). Links Hinoda, Y., Neumaier, M., Hefta, S. A., Drzeniek, Z., Wagner, C, Shively, L., Hefta, L. J. F., Shively, J. E. and Paxton, R. J., Molecular cloning of a cDNA coding biliary glycoprotein. I: Primary structure of a glycoprotein immunologically cross-reactive with carcinoembryonic antigen. Proc. nat. Acad. Sci. (Wash.) , 85, 6959-6963 (1988). Links Khan, W. N., Frängsmyr, L., Teglund, S., Israelsson, A., Bremer, K. and Hammarström, S., Identification of three new genes and estimation of the size of the carcinoembryonic antigen family. Genomics , 14, 384-390 (1992). Links Kim, J., Kaye, F. J., Henslee, J. G., Shively, J. E., Park, J.-G., Lai, S.-L., Linnoila, R. I., Mulshine, J. L. and Gazdar, A. F., Expression of carcinoembryonic antigen and related genes in lung and gastrointestinal cancers. Int. J. Cancer , 52, 718-725 (1992). Links Kuroki, M., Arakawa, F., Matsuo, Y., Oikawa, S., Misumi, Y., Nakazato, H. and Matsuoka, Y., Molecular cloning of non-specific cross-reacting antigens in human granulocytes. J. biol. Chem. , 266, 11810-11817 (1991). Links Lathe, R., Synthetic oligonucleotide probes deduced from amino acid sequence data: theoretical and practical considerations. J. mol. Biol. , 183, 1-12 (1985). Links Lucas, K., Busch, M., Mössinger, S. and Thompson, J. A., An improved microcomputer program for finding gene- or gene-family-specific oligonucleotides suitable as primers for polymerase chain reactions or as probes. CABIOS , 7, 525-529 (1991). Links Motoyama, T., Watanabe, H., Takeuchi, S., Watanabe, T., Gotoh, S. and Okazaki, E., Cancer antigen 125, carcinoembryonic antigen, and carbohydrate determinant 19-9 in ovarian tumors. Cancer , 66, 2628-2635 (1990). Links Nagel, G., Grunert, F., Kuijpers, T. W., Watt, S., Thompson, J. and Zimmermann, W., Genomic organization, splice variants and expression of CGM1, a CD66-related member of the carcinoembryonic-antigen-gene family. Europ. J. Biochem. , 214, 27-35 (1993). Links Neumaier, M., Zimmermann, W., Shively, L., Hinoda, Y., Riggs, A. D. and Shively, J. E., Characterization of a cDNA clone for the non-specific cross-reacting antigen (NCA) and a comparison of NCA and carcinoembryonic antigen (CEA). J. biol. Chem. , 263, 3202-3207 (1988). Links Ng, S. Y., Gunning, P., Eddy, R., Ponte, P., Leavitt, J., Shows, T. and Kedes, L., Evolution of the functional human -actin gene and its multi-pseudogene family: conservation of non-coding regions and chromosomal dispersion of pseudosenes. Mol. cell. Biol , 5, 2720-2732 (1985). Links Oikawa, S., Nakazato, H. and Kosaki, G., Primary structure of human carcinoembryonic antigen (CEA) deduced from cDNA sequence. Biochem. biophys. Res. Commun. , 142, 511-518 (1987). Links Pèlegrin, A., Terskikh, A., Hayoz, D., Chalandon, Y., Olsson, N.-O., Folli, S., Buchegger, F., Kromer, B., Schwarz, K., Martin, M., Martin, F. and Mach, J.-P., Human carcinoembryonic antigen cDNA expressed in rat carcinoma cells can function as a target antigen for tumor localization of antibodies in nude rats and as rejection antigen in syngeneic rats. Int. J. Cancer , 52, 110-119 (1992). Links Ponte, P., Ng, S.-Y., Engel, J., Gunning, P. and Kedes, L., Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta-actin cDNA. Nucl. Acids Res. , 12, 1687-1696 (1984). Links Rooney, B. C., Horne, C. H. W. and Hardman, N., Molecular cloning of a cDNA for human pregnancy-specific 1-glycoprotein: homology with human carcinoembryonic antigen and related proteins. Gene , 71, 439-449 (1988). Links Sakuragi, N., Serum SP1 and hCG beta sub-unit (hCG beta) levels in choriocarcinoma, invasive mole, and hydatidiform mole: clinical significance of SP1/hCG beta ratio. Gynecol. Oncol , 13, 393-398 (1982). Links Sambrook, J. E., Fritsch, J. and Maniatis, T., Molecular cloning: a laboratory manual (2nd ed.), Cold Spring Harbor Laboratory, New York (1989). Shively, J. E. and Beatty, J. D., CEA-related antigens: molecular biology and clinical significance. CRC Crit. Rev. Oncol. Hematol , 2, 355-399 (1985). Links Thompson, J. A., Grunert, F. and Zimmermann, W., The carcinoem-bryonic-antigen-gene family: molecular biology and clinical perspectives. J clin. Lab. Analysis , 5, 344-366 (1991). Links