Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

A single-injection protein kinase A-directed antisense treatment to inhibit tumour growth

Abstract

Expression of the RIα subunit of cAMP-dependent protein kinase type I is enhanced in human cancer cell lines, in primary tumours, in cells after transformation and in cells upon stimulation of growth. We have investigated the effect of sequence-specific inhibition of RIα gene expression on in vivo tumour growth. We report that single injection RIα antisense treatment results in a reduction in RIα expression and inhibition of tumour growth. Tumour cells behaved like untransformed cells by making less protein kinase type I. The RIα antisense, which produces a biochemical imprint for growth control, requires infrequent dosing to halt neoplastic growth in vivo.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Krebs, E.G. Protein kinase. Curr. Topics Cell Regul. 5, 99–133 (1972).

    Article  CAS  Google Scholar 

  2. Lohmann, S.M. & Walter, U. Regulation of the cellular and subcellular concentrations and distribution of cyclic nucleotide-dependent protein kinases. in Advances in Cyclic Nucleotide and Protein Phosphorylation Research, vol. 18, 63–117 (Raven, New York, 1984).

    Google Scholar 

  3. Cho-Chung, Y.S. Role of cyclic AMP receptor proteins in growth, differentiation, and suppression of malignancy: New approaches to therapy [Perspectives in cancer research]. Cancer Res. 50, 7093–7100 (1990).

    CAS  PubMed  Google Scholar 

  4. Beebe, S.J. & Corbin, J.D. Cyclic nucleotide-dependent protein kinases. in The Enzymes: Control by Phosphorylation, vol. 17, part A, 43–111 (Academic, New York, 1986).

    Google Scholar 

  5. McKnight, G.S. et al. Analysis of the cAMP-dependent protein kinase system using molecular genetic approaches. Recent Prog. Horm. Res. 44, 307–335 (1988).

    CAS  PubMed  Google Scholar 

  6. Levy, F.O. et al. Molecular cloning, complementary deoxyribonucleic acid structure and predicted full-length amino acid sequence of the hormone-inducible regulatory subunit of 3′,5′-cyclic adenoslne monophosphate-dependent protein kinase from human testis. Molec. Endocrinol. 2, 1364–1373 (1988).

    Article  CAS  Google Scholar 

  7. Uhler, M.D. et al. Isolation of cDNA clones coding for the catalytic subunit of mouse cAMP-dependent protein kinase. Proc. natn. Acad. Sci. U.S.A. 83, 1300–1304 (1986).

    Article  CAS  Google Scholar 

  8. Uhler, M.D., Chrivia, J.C. & McKnight, G.S. Evidence for a second isoform of the catalytic subunit of cAMP-dependent protein kinase. J. biol. Chem. 261, 15360–15363 (1986).

    CAS  PubMed  Google Scholar 

  9. Showers, M.O. & Maurer, R.A. A cloned bovine cDNA encodes an alternate form of the catalytic subunit of cAMP-dependent protein kinase. J. biol. Chem. 261, 16288–16291 (1986).

    CAS  PubMed  Google Scholar 

  10. Beebe, S.J. et al. Molecular cloning of a unique tissue-specific protein kinase (Cγ) from human testis—representing a third isoform for the catalytic subunit of the cAMP-dependent protein kinase. Molec. Endocrinol. 4, 465–475 (1990).

    Article  CAS  Google Scholar 

  11. Øyen, O. et al. A unique mRNA species for a regulatory subunit of cAMP-dependent protein kinase is specifically induced in haploid germ cells. FEBS Lett. 229, 391–394 (1988).

    Article  Google Scholar 

  12. Clegg, C.H., Cadd, G.G. & McKnight, G.S. Genetic characterization of a brain-specific form of the type I regulatory subunit of cAMP-dependent protein kinase. Proc. natn. Acad. Sci. U.S.A. 85, 3703–3707 (1988).

    Article  CAS  Google Scholar 

  13. Cadd, G.G., Uhler, M.D. & McKnight, G.S. Holoenzymes of cAMP-dependent protein kinase containing the neural form of type I regulatory subunit have an increased sensitivity to cyclic nucleotides. J. biol. Chem. 265, 19502–19506 (1990).

    CAS  PubMed  Google Scholar 

  14. Kapoor, C.L. & Cho-Chung, Y.S. Compartmentalization of regulatory subunits of cyclic adenosine 3′, 5′-monophosphate-dependent protein kinases in MCF-7 human breast cancer cells. Cancer Res. 43, 295–302 (1983).

    CAS  PubMed  Google Scholar 

  15. Nigg, E.A., Schäfer, G., Hilz, H. & Eppenberger, H.M. Cyclic-AMP-dependent protein kinase type II is associated with the golgi complex and with centrosomes. Cell 41, 1039–1051 (1985).

    Article  CAS  Google Scholar 

  16. Taylor, S.S. et al. cAMP-dependent protein kinase: Prototype for a family of enzymes. FASEB J. 2, 2677–2685 (1988).

    Article  CAS  Google Scholar 

  17. Yokozaki, H. et al. An antisense oligodeoxynucleotide that depletes RIα sub-unit of cyclic AMP-dependent protein kinase induces growth inhibition in human cancer cells. Cancer Res. 53, 868–872 (1993).

    CAS  PubMed  Google Scholar 

  18. Tortora, G., Yokozaki, H., Pepe, S., Clair, T. & Cho-Chung, Y.S. Differentiation of HL-60 leukemia by type I regulatory subunit antisense oligodeoxynucleotide of cAMP-dependent protein kinase. Proc. natn. Acad. Sci. U.S.A. 88, 2011–2015 (1991).

    Article  CAS  Google Scholar 

  19. Sandberg, M., Tasken, K., Øyen, O., Hansson, V. & Jahnsen, T. Molecular cloning, cDNA structure and deduced amino acid sequence for A type I regulatory subunit of cAMP-dependent protein kinase from human testis. Biochem. biophys. Res. Commun. 149, 939–945 (1987).

    Article  CAS  Google Scholar 

  20. Tortora, G., Clair, T. & Cho-Chung, Y.S. An antisense oligodeoxynucleotide targeted against the type RIIβ, regulatory subunit mRNA of protein kinase inhibits cAMP-induced differentiation in HL-60 leukemia Cells without affecting phorbol ester effects. Proc. natn. Acad. Sci. U.S.A. 87, 705–708 (1990).

    Article  CAS  Google Scholar 

  21. Rosen, O.M. & Erlichman, J. Reversible autophosphorylation of a cyclic 3′, 5′-AMP-dependent protein kinase from bovine cardiac muscle. J. biol. Chem. 250, 7788–7794 (1975).

    CAS  PubMed  Google Scholar 

  22. Rohlff, C., Clair, T. & Cho-Chung, Y.S. 8-Cl-cAMP induces truncation and down-regulation of the RIα subunit and up-regulation of the RIIβ, subunit of cAMP-dependent protein kinase leading to type II holoenzyme-dependent growth inhibition and differentiation of HL-60 leukemia cells. J. biol. Chem. 268, 5774–5782 (1993).

    CAS  PubMed  Google Scholar 

  23. Tagliaferri, P., Katsaros, D., Clair, T., Neckers, L., Robins, R.K. & Cho-Chung, Y.S. Reverse transformation of Harvey murine sarcoma virus-transformed NIH/3T3 cells by site-selective cyclic AMP analogs. J. biol. Chem. 263, 409–416 (1988).

    CAS  PubMed  Google Scholar 

  24. Connelly, P.A., Hastings, T.G. & Reimann, E.M. Identification of a ternary complex between cAMP and a trimeric form of cAMP-dependent protein kinase. J. biol. Chem. 261, 2325–2330 (1986).

    CAS  PubMed  Google Scholar 

  25. Cobb, C.E., Beth, A.H. & Corbin, J.D. Purification and characterization of an inactive form of cAMP-dependent protein kinase containing bound cAMP. J. biol. Chem. 262, 16566–16574 (1987).

    CAS  PubMed  Google Scholar 

  26. Otten, A.D., Parenteau, L.A., Døskeland, S. & McKnight, G.S. Hormonal activation of gene transcription in ras-transformed NIH3T3 cells overexpressing RIIα and RIIβ subunits of the cAMP-dependent protein kinase. J. biol. Chem. 266, 23074–23082 (1991).

    CAS  PubMed  Google Scholar 

  27. Nesterova, M., Budillon, A., Pepe, S., Cereseto, A. & Cho-Chung, Y.S. Introducing an autophosphorylation site mutation in the RIIβ regulatory subunit of cAMP-dependent protein kinase abolishes the RIIβ-mediated regulatory function. FASEB J. 8, A1328 (1994) (abstract).

    Google Scholar 

  28. Srivastava, A.K. & Stellwagen, R.H. Presence of the sites for interacting with cyclic AMP and with catalytic subunit on small fragments of protein kinase regulatory subunit. J. biol. Chem. 253, 1752–1755 (1978).

    CAS  PubMed  Google Scholar 

  29. Wagner, R.W. Gene inhibition using antisense oligodeoxynucleotides. Nature 372, 333–335 (1994).

    Article  CAS  Google Scholar 

  30. Iversen, P. In vivo studies with phosphorothioate oligonucleotides: Pharmacokinetics prologue. Anti-Cancer Drug Des. 6, 531–538 (1991).

    CAS  Google Scholar 

  31. Weber, W. & Hilz, H. cAMP-dependent protein kinases I and II: Divergent turnover of subunits. Biochemistry 25, 5661–5667 (1986).

    Article  CAS  Google Scholar 

Download references

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nesterova, M., Cho-Chung, Y. A single-injection protein kinase A-directed antisense treatment to inhibit tumour growth. Nat Med 1, 528–533 (1995). https://doi.org/10.1038/nm0695-528

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm0695-528

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing