Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1989 Sep 15;262(3):873–879. doi: 10.1042/bj2620873

Metallothionein mRNA and protein induction by cadmium in peripheral-blood leucocytes.

C B Harley 1, C R Menon 1, R A Rachubinski 1, E Nieboer 1
PMCID: PMC1133355  PMID: 2590172

Abstract

To help to evaluate the role of metallothionein (MT) in peripheral-blood leucocytes, we examined MT protein and mRNA levels in these cells before and after exposure to CdCl2 in culture. Protein was assayed by 109Cd2+ binding, and RNA by dot-blot hybridization. MT was induced in both lymphocytes and adherent monocytes about 10-fold with a 12 h exposure to 10 microM-CdCl2, but absolute levels were 3-fold higher in monocytes: 57 x 10(5) (+Cd2+) versus 6 x 10(5) (-Cd2+) molecules/cell for monocytes; 18 x 10(5) (+Cd2+) versus 2 x 10(5) (-Cd2+) for lymphocytes. Polymorphonuclear cells expressed relatively little MT (0.6 x 10(5) molecules/cell), and this did not change with phorbol ester stimulation or exposure to Cd2+, arguing against a direct protective role for MT in activated neutrophils. MT mRNA levels corresponded qualitatively to expression of protein in these cells. Our data provide quantitative comparisons of leucocyte MT expression and regulation in the human population. Variation in both basal and induced MT mRNA levels reflects environmental or experimental (intra-individual) and possibly genetic (inter-individual) differences.

Full text

PDF
873

Images in this article

Selected References

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

  1. Bell T. V., Harley C. B., Stetsko D., Sauder D. N. Expression of mRNA homologous to interleukin 1 in human epidermal cells. J Invest Dermatol. 1987 Apr;88(4):375–379. doi: 10.1111/1523-1747.ep12469041. [DOI] [PubMed] [Google Scholar]
  2. Dieter H. H., Müller L., Abel J., Summer K. H. Determination of Cd-thionein in biological materials: comparative standard recovery by five current methods using protein nitrogen for standard calibration. Toxicol Appl Pharmacol. 1986 Sep 30;85(3):380–388. doi: 10.1016/0041-008x(86)90345-5. [DOI] [PubMed] [Google Scholar]
  3. Durnam D. M., Hoffman J. S., Quaife C. J., Benditt E. P., Chen H. Y., Brinster R. L., Palmiter R. D. Induction of mouse metallothionein-I mRNA by bacterial endotoxin is independent of metals and glucocorticoid hormones. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1053–1056. doi: 10.1073/pnas.81.4.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Durnam D. M., Palmiter R. D. Transcriptional regulation of the mouse metallothionein-I gene by heavy metals. J Biol Chem. 1981 Jun 10;256(11):5712–5716. [PubMed] [Google Scholar]
  5. Friedman R. L., Manly S. P., McMahon M., Kerr I. M., Stark G. R. Transcriptional and posttranscriptional regulation of interferon-induced gene expression in human cells. Cell. 1984 Oct;38(3):745–755. doi: 10.1016/0092-8674(84)90270-8. [DOI] [PubMed] [Google Scholar]
  6. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  7. Hager L. J., Palmiter R. D. Transcriptional regulation of mouse liver metallothionein-I gene by glucocorticoids. Nature. 1981 May 28;291(5813):340–342. doi: 10.1038/291340a0. [DOI] [PubMed] [Google Scholar]
  8. Hamer D. H. Metallothionein. Annu Rev Biochem. 1986;55:913–951. doi: 10.1146/annurev.bi.55.070186.004405. [DOI] [PubMed] [Google Scholar]
  9. Hanke T., Tyers M., Harley C. B. Metallothionein RNA levels in HL-60 cells. Effect of cadmium, differentiation, and protein kinase C activation. FEBS Lett. 1988 Dec 5;241(1-2):159–163. doi: 10.1016/0014-5793(88)81051-2. [DOI] [PubMed] [Google Scholar]
  10. Harley C. B. Hybridization of oligo(dT) to RNA on nitrocellulose. Gene Anal Tech. 1987 Mar-Apr;4(2):17–22. doi: 10.1016/0735-0651(87)90013-6. [DOI] [PubMed] [Google Scholar]
  11. Harley C. B., Pollard J. W., Chamberlain J. W., Stanners C. P., Goldstein S. Protein synthetic errors do not increase during aging of cultured human fibroblasts. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1885–1889. doi: 10.1073/pnas.77.4.1885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hildebrand C. E., Cram L. S. Distribution of cadmium in human blood cultured in low levels of CdCl2: accumulation of Cd in lymphocytes and preferential binding to metallothionein. Proc Soc Exp Biol Med. 1979 Sep;161(4):438–443. doi: 10.3181/00379727-161-40569. [DOI] [PubMed] [Google Scholar]
  13. Imbra R. J., Karin M. Metallothionein gene expression is regulated by serum factors and activators of protein kinase C. Mol Cell Biol. 1987 Apr;7(4):1358–1363. doi: 10.1128/mcb.7.4.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jin T., Nordberg G. F., Nordberg M. Influence of cadmium-metallothionein pretreatment on tolerance of rat kidney cortical cells to cadmium toxicity in vitro and in vivo. Pharmacol Toxicol. 1987 May;60(5):345–349. doi: 10.1111/j.1600-0773.1987.tb01524.x. [DOI] [PubMed] [Google Scholar]
  15. Karin M., Haslinger A., Holtgreve H., Cathala G., Slater E., Baxter J. D. Activation of a heterologous promoter in response to dexamethasone and cadmium by metallothionein gene 5'-flanking DNA. Cell. 1984 Feb;36(2):371–379. doi: 10.1016/0092-8674(84)90230-7. [DOI] [PubMed] [Google Scholar]
  16. Karin M., Imbra R. J., Heguy A., Wong G. Interleukin 1 regulates human metallothionein gene expression. Mol Cell Biol. 1985 Oct;5(10):2866–2869. doi: 10.1128/mcb.5.10.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koropatnick J., Winning R., Wiese E., Heschl M., Gedamu L., Duerksen J. Acute treatment of mice with cadmium salts results in amplification of the metallothionein-1 gene in liver. Nucleic Acids Res. 1985 Aug 12;13(15):5423–5439. doi: 10.1093/nar/13.15.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. MacArthur C. A., Lieberman M. W. Different types of hypersensitive sites in the mouse metallothionein gene region. J Biol Chem. 1987 Feb 15;262(5):2161–2165. [PubMed] [Google Scholar]
  19. Matsubara J., Tajima Y., Karasawa M. Metallothionein induction as a potent means of radiation protection in mice. Radiat Res. 1987 Aug;111(2):267–275. [PubMed] [Google Scholar]
  20. Monia B. P., Butt T. R., Ecker D. J., Mirabelli C. K., Crooke S. T. Metallothionein turnover in mammalian cells. Implications in metal toxicity. J Biol Chem. 1986 Aug 25;261(24):10957–10959. [PubMed] [Google Scholar]
  21. Morgan A. R., Lee J. S., Pulleyblank D. E., Murray N. L., Evans D. H. Review: ethidium fluorescence assays. Part 1. Physicochemical studies. Nucleic Acids Res. 1979 Oct 10;7(3):547–569. doi: 10.1093/nar/7.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nolan C. V., Shaikh Z. A. Determination of metallothionein in tissues by radioimmunoassay and by cadmium saturation method. Anal Biochem. 1986 Apr;154(1):213–223. doi: 10.1016/0003-2697(86)90518-x. [DOI] [PubMed] [Google Scholar]
  23. Onosaka S., Cherian M. G. Comparison of metallothionein determination by polarographic and cadmium-saturation methods. Toxicol Appl Pharmacol. 1982 Apr;63(2):270–274. doi: 10.1016/0041-008x(82)90049-7. [DOI] [PubMed] [Google Scholar]
  24. Patierno S. R., Costa M., Lewis V. M., Peavy D. L. Inhibition of LPS toxicity for macrophages by metallothionein-inducing agents. J Immunol. 1983 Apr;130(4):1924–1929. [PubMed] [Google Scholar]
  25. Peavy D. L., Fairchild E. J., 2nd Induction of metallothionein synthesis in human peripheral blood leukocytes. Environ Res. 1987 Apr;42(2):377–385. doi: 10.1016/s0013-9351(87)80204-9. [DOI] [PubMed] [Google Scholar]
  26. Roels H., Lauwerys R., Buchet J. P., Bernard A., Garvey J. S., Linton H. J. Significance of urinary metallothionein in workers exposed to cadmium. Int Arch Occup Environ Health. 1983;52(2):159–166. doi: 10.1007/BF00405419. [DOI] [PubMed] [Google Scholar]
  27. Schmidt C. J., Hamer D. H. Cell specificity and an effect of ras on human metallothionein gene expression. Proc Natl Acad Sci U S A. 1986 May;83(10):3346–3350. doi: 10.1073/pnas.83.10.3346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sone T., Yamaoka K., Minami Y., Tsunoo H. Induction of metallothionein synthesis in Menkes' and normal lymphoblastoid cells is controlled by the level of intracellular copper. J Biol Chem. 1987 Apr 25;262(12):5878–5885. [PubMed] [Google Scholar]
  29. Thomas J. P., Bachowski G. J., Girotti A. W. Inhibition of cell membrane lipid peroxidation by cadmium- and zinc-metallothioneins. Biochim Biophys Acta. 1986 Dec 10;884(3):448–461. doi: 10.1016/0304-4165(86)90195-9. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES