Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1985 Mar;4(3):715–724. doi: 10.1002/j.1460-2075.1985.tb03688.x

Expression of the human calcitonin/CGRP gene in lung and thyroid carcinoma.

M R Edbrooke, D Parker, J H McVey, J H Riley, G D Sorenson, O S Pettengill, R K Craig
PMCID: PMC554247  PMID: 2408883

Abstract

Nucleotide sequence analysis of a partially processed polyadenylated precursor RNA transcript shows that the human calcitonin gene in common with the rat calcitonin gene, encodes calcitonin and the calcitonin gene related peptide (CGRP). Using hybridisation probes specific to calcitonin mRNA, intron, coding and non-coding regions of the CGRP mRNA, we demonstrate by Southern blotting the existence of a second human CGRP gene, and by RNA blotting and S1 mapping, the differential expression of calcitonin and CGRP in medullary thyroid carcinoma and human lung tumour cell-lines. These studies implicate the requirement for separate post-transcriptional events in the differential expression of calcitonin and CGRP from a single gene, the preferential use of splice acceptor sites for the synthesis of CGRP mRNA, and post-transcriptional cleavage modulated by a trans-acting gene product for the synthesis of calcitonin mRNA. Studies using antisera raised against CGRP and calcitonin, demonstrate elevated circulating levels of plasma CGRP in medullary thyroid carcinoma which do not parallel calcitonin levels, and the presence of CGRP in secretions from lung tumour cell-lines. These studies indicate that CGRP is a tumour marker of diagnostic and possibly prognostic value in the management of lung and thyroid tumours.

Full text

PDF
715

Images in this article

718Fig. 2.
on p.718
719Fig. 3.
on p.719
720Fig. 4.
on p.720

Selected References

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

  1. Allison J., Hall L., MacIntyre I., Craig R. K. The construction and partial characterization of plasmids containing complementary DNA sequences to human calcitonin precursor polyprotein. Biochem J. 1981 Dec 1;199(3):725–731. doi: 10.1042/bj1990725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amara S. G., Jonas V., Rosenfeld M. G., Ong E. S., Evans R. M. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature. 1982 Jul 15;298(5871):240–244. doi: 10.1038/298240a0. [DOI] [PubMed] [Google Scholar]
  3. Austin L. A., Heath H., 3rd Calcitonin: physiology and pathophysiology. N Engl J Med. 1981 Jan 29;304(5):269–278. doi: 10.1056/NEJM198101293040505. [DOI] [PubMed] [Google Scholar]
  4. Berget S. M. Are U4 small nuclear ribonucleoproteins involved in polyadenylation? Nature. 1984 May 10;309(5964):179–182. doi: 10.1038/309179a0. [DOI] [PubMed] [Google Scholar]
  5. Birchmeier C., Folk W., Birnstiel M. L. The terminal RNA stem-loop structure and 80 bp of spacer DNA are required for the formation of 3' termini of sea urchin H2A mRNA. Cell. 1983 Dec;35(2 Pt 1):433–440. doi: 10.1016/0092-8674(83)90176-9. [DOI] [PubMed] [Google Scholar]
  6. Bradbury A. F., Finnie M. D., Smyth D. G. Mechanism of C-terminal amide formation by pituitary enzymes. Nature. 1982 Aug 12;298(5875):686–688. doi: 10.1038/298686a0. [DOI] [PubMed] [Google Scholar]
  7. Coombes R. C., Hillyard C., Greenberg P. B., MacIntyre I. Plasma-immunoreactive-calcitonin in patients with non-thyroid tumours. Lancet. 1974 Jun 1;1(7866):1080–1083. doi: 10.1016/s0140-6736(74)90557-1. [DOI] [PubMed] [Google Scholar]
  8. Craig R. K., Brown P. A., Harrison O. S., McIlreavy D., Campbell P. N. Guinea-pig milk-protein synthesis. Isolation and characterization of messenger ribonucleic acids from lactating mammary gland and identification of caseins and pre-alpha-lactalbumin as translation products in heterologous cell-free systems. Biochem J. 1976 Oct 15;160(1):57–74. doi: 10.1042/bj1600057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Craig R. K., Hall L., Edbrooke M. R., Allison J., MacIntyre I. Partial nucleotide sequence of human calcitonin precursor mRNA identifies flanking cryptic peptides. Nature. 1982 Jan 28;295(5847):345–347. doi: 10.1038/295345a0. [DOI] [PubMed] [Google Scholar]
  10. Fisher L. A., Kikkawa D. O., Rivier J. E., Amara S. G., Evans R. M., Rosenfeld M. G., Vale W. W., Brown M. R. Stimulation of noradrenergic sympathetic outflow by calcitonin gene-related peptide. Nature. 1983 Oct 6;305(5934):534–536. doi: 10.1038/305534a0. [DOI] [PubMed] [Google Scholar]
  11. Galli G., Hofstetter H., Stunnenberg H. G., Birnstiel M. L. Biochemical complementation with RNA in the Xenopus oocyte: a small RNA is required for the generation of 3' histone mRNA termini. Cell. 1983 Oct;34(3):823–828. doi: 10.1016/0092-8674(83)90539-1. [DOI] [PubMed] [Google Scholar]
  12. Gil A., Proudfoot N. J. A sequence downstream of AAUAAA is required for rabbit beta-globin mRNA 3'-end formation. 1984 Nov 29-Dec 5Nature. 312(5993):473–474. doi: 10.1038/312473a0. [DOI] [PubMed] [Google Scholar]
  13. Girgis S. I., Galan F. G., Arnett T. R., Rogers R. M., Bone Q., Ravazzola M., MacIntyre I. Immunoreactive human calcitonin-like molecule in the nervous systems of protochordates and a cyclostome, Myxine. J Endocrinol. 1980 Dec;87(3):375–382. doi: 10.1677/joe.0.0870375. [DOI] [PubMed] [Google Scholar]
  14. Hall L., Craig R. K., Campbell P. N. mRNA species directing synthesis of milk proteins in normal and tumour tissue from human mammary gland. Nature. 1979 Jan 4;277(5691):54–56. doi: 10.1038/277054a0. [DOI] [PubMed] [Google Scholar]
  15. Kühne T., Wieringa B., Reiser J., Weissmann C. Evidence against a scanning model of RNA splicing. EMBO J. 1983;2(5):727–733. doi: 10.1002/j.1460-2075.1983.tb01492.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Le Moullec J. M., Jullienne A., Chenais J., Lasmoles F., Guliana J. M., Milhaud G., Moukhtar M. S. The complete sequence of human preprocalcitonin. FEBS Lett. 1984 Feb 13;167(1):93–97. doi: 10.1016/0014-5793(84)80839-x. [DOI] [PubMed] [Google Scholar]
  17. Lippman S. M., Mendelsohn G., Trump D. L., Wells S. A., Jr, Baylin S. B. The prognostic and biological significance of cellular heterogeneity in medullary thyroid carcinoma: a study of calcitonin, L-dopa decarboxylase, and histaminase. J Clin Endocrinol Metab. 1982 Feb;54(2):233–240. doi: 10.1210/jcem-54-2-233. [DOI] [PubMed] [Google Scholar]
  18. Mather E. L., Nelson K. J., Haimovich J., Perry R. P. Mode of regulation of immunoglobulin mu- and delta-chain expression varies during B-lymphocyte maturation. Cell. 1984 Feb;36(2):329–338. doi: 10.1016/0092-8674(84)90226-5. [DOI] [PubMed] [Google Scholar]
  19. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  20. Mendelsohn G., Wells S. A., Jr, Baylin S. B. Relationship of tissue carcinoembryonic antigen and calcitonin to tumor virulence in medullary thyroid carcinoma. An immunohistochemical study in early, localized, and virulent disseminated stages of disease. Cancer. 1984 Aug 15;54(4):657–662. doi: 10.1002/1097-0142(1984)54:4<657::aid-cncr2820540412>3.0.co;2-v. [DOI] [PubMed] [Google Scholar]
  21. Milhaud G., Calmette C., Taboulet J., Julienne A., Moukhtar M. S. Letter: Hypersecretion of calcitonin in neoplastic conditions. Lancet. 1974 Mar 16;1(7855):462–463. doi: 10.1016/s0140-6736(74)92428-3. [DOI] [PubMed] [Google Scholar]
  22. Morris H. R., Panico M., Etienne T., Tippins J., Girgis S. I., MacIntyre I. Isolation and characterization of human calcitonin gene-related peptide. Nature. 1984 Apr 19;308(5961):746–748. doi: 10.1038/308746a0. [DOI] [PubMed] [Google Scholar]
  23. Nelkin B. D., Rosenfeld K. I., de Bustros A., Leong S. S., Roos B. A., Baylin S. B. Structure and expression of a gene encoding human calcitonin and calcitonin gene related peptide. Biochem Biophys Res Commun. 1984 Sep 17;123(2):648–655. doi: 10.1016/0006-291x(84)90278-x. [DOI] [PubMed] [Google Scholar]
  24. Pettengill O. S., Sorenson G. D., Wurster-Hill D. H., Curphey T. J., Noll W. W., Cate C. C., Maurer L. H. Isolation and growth characteristics of continuous cell lines from small-cell carcinoma of the lung. Cancer. 1980 Mar 1;45(5):906–918. doi: 10.1002/1097-0142(19800301)45:5<906::aid-cncr2820450513>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  25. Reichlin M. Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods Enzymol. 1980;70(A):159–165. doi: 10.1016/s0076-6879(80)70047-2. [DOI] [PubMed] [Google Scholar]
  26. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  27. Roos B. A., Huber M. B., Birnbaum R. S., Aron D. C., Lindall A. W., Lips K., Baylin S. B. Medullary thyroid carcinomas secrete a noncalcitonin peptide corresponding to the carboxyl-terminal region of preprocalcitonin. J Clin Endocrinol Metab. 1983 Apr;56(4):802–807. doi: 10.1210/jcem-56-4-802. [DOI] [PubMed] [Google Scholar]
  28. Rosenfeld M. G., Amara S. G., Evans R. M. Alternative RNA processing: determining neuronal phenotype. Science. 1984 Sep 21;225(4668):1315–1320. doi: 10.1126/science.6089345. [DOI] [PubMed] [Google Scholar]
  29. Rosenfeld M. G., Mermod J. J., Amara S. G., Swanson L. W., Sawchenko P. E., Rivier J., Vale W. W., Evans R. M. Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature. 1983 Jul 14;304(5922):129–135. doi: 10.1038/304129a0. [DOI] [PubMed] [Google Scholar]
  30. Steenbergh P. H., Höppener J. W., Zandberg J., Van de Ven W. J., Jansz H. S., Lips C. J. Calcitonin gene related peptide coding sequence is conserved in the human genome and is expressed in medullary thyroid carcinoma. J Clin Endocrinol Metab. 1984 Aug;59(2):358–360. doi: 10.1210/jcem-59-2-358. [DOI] [PubMed] [Google Scholar]
  31. Stunnenberg H. G., Birnstiel M. L. Bioassay for components regulating eukaryotic gene expression: a chromosomal factor involved in the generation of histone mRNA 3' termini. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6201–6204. doi: 10.1073/pnas.79.20.6201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Taya Y., Devos R., Tavernier J., Cheroutre H., Engler G., Fiers W. Cloning and structure of the human immune interferon-gamma chromosomal gene. EMBO J. 1982;1(8):953–958. doi: 10.1002/j.1460-2075.1982.tb01277.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Taylor J. B., Craig R. K., Beale D., Ketterer B. Construction and characterization of a plasmid containing complementary DNA to mRNA encoding the N-terminal amino acid sequence of the rat glutathione transferase Ya subunit. Biochem J. 1984 Apr 1;219(1):223–231. doi: 10.1042/bj2190223. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES