Research Article | Published:

Induction of endogenous Bcl-xS through the control of Bcl-x pre-mRNA splicing by antisense oligonucleotides

Nature Biotechnology volume 17, pages 10971100 (1999) | Download Citation

Subjects

Abstract

Resistance to apoptosis, which plays an important role in tumors that are refractory to chemotherapy, is regulated by the ratio of antiapoptotic to proapoptotic proteins. By manipulating levels of these proteins, cells can become sensitized to undergo apoptosis in response to chemotherapeutic agents. Alternative splicing of the bcl-x gene gives rise to two proteins with antagonistic functions: Bcl-xL, a well-characterized antiapoptotic protein, and Bcl-xS, a proapoptotic protein. We show here that altering the ratio of Bcl-xL to Bcl-xS in the cell using an antisense oligonucleotide permitted cells to be sensitized to undergo apoptosis in response to ultraviolet B radiation and chemotherapeutic drug treatment. These results demonstrate the ability of a chemically modified oligonucleotide to alter splice site selection in an endogenous gene and illustrate a powerful tool to regulate cell survival.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Cell suicide for beginners. Nature 396, 119– 122 (1998).

  2. 2.

    Life, death, and the pursuit of apoptosis. Genes Dev. 10, 1–15 (1996).

  3. 3.

    & Alternative splicing and programmed cell death. Proc. Soc. Exp. Biol. Med. 220, 64–72 (1999).

  4. 4.

    et al. Bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 74, 597– 608 (1993).

  5. 5.

    & Molecular thanatopsis: a discourse on the BCL2 family and cell death. Blood 88, 386–401 (1996).

  6. 6.

    & Bcl-2-family proteins: the role of the BH3 domain in apoptosis. Trends Cell Biol. 8 , 324–330 (1998).

  7. 7.

    et al. Bcl-x is expressed in embryonic and postnatal neural tissues and functions to prevent neuronal cell death. Proc. Natl. Acad. Sci. USA 92, 4304–4308 (1995).

  8. 8.

    et al. An additional form of rat Bcl-x, Bcl-xβ, generated by an unspliced RNA, promotes apoptosis in promyeloid cells. J. Biol. Chem. 271, 13258–13265 (1996).

  9. 9.

    , , , & Identification of a human cDNA encoding a novel Bcl-x isoform. Biochem. Biophys. Res. Commun. 248, 147–152 (1998).

  10. 10.

    Regulation of apoptosis by bcl-2 family proteins and its role in cancer chemoresistance. Curr. Opin. Oncol. 7, 541– 546 (1995).

  11. 11.

    & Regulation of pre-mRNA splicing by antisense oligonucleotides. Curr. Opin. Drug Discovery Dev. 2, 147–151 (1999).

  12. 12.

    Modification of pre-mRNA splicing by antisense oligonucleotides. Acta Biochim Pol. 44, 231–238 (1997).

  13. 13.

    & Inhibition of splicing of wild-type and mutated luciferase adenovirus pre-mRNAs by antisense oligonucleotides. Mol. Pharmacol. 48, 905– 918 (1995).

  14. 14.

    & Restoration of correct splicing in thalassemic pre-mRNA by antisense oligonucleotides. Proc. Natl. Acad. Sci. USA 90, 8673–8677 (1993).

  15. 15.

    , , & Repair of thalassemic human β-globin mRNA in mammalian cells by antisense oligonucleotides. Proc. Natl. Acad. Sci. USA 93, 12840– 12844 (1996).

  16. 16.

    , , , & Modification of splicing in the dystrophin gene in cultured Mdx muscle cells by antisense oligoribonucleotides. Hum. Mol. Genet. 5, 1083– 1090 (1998).

  17. 17.

    & Identification and characterization of second-generation antisense oligonucleotides. Antisense Nucleic Acid Drug Dev. 7, 229–233 (1997).

  18. 18.

    , , , & Inhibition of Bcl-xL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli. Oncogene 18, 4495–4504 (1999).

  19. 19.

    , , & Effects of exogenous wild-type p53 on a human lung carcinoma cell line with endogenous wild-type p53. Exp. Cell Res. 203, 297–304 (1992).

  20. 20.

    et al. Ras-dependent apoptosis correlates with persistent activation of stress-activated protein kinases and induction of isoform(s) of Bcl-x. Cell Death Differ. 4, 745–755 (1997).

  21. 21.

    et al. A recombinant bcl-xS adenovirus selectively induces apoptosis in cancer cells but not in normal bone marrow cells. Proc. Natl. Acad. USA 92, 11024–11028 (1995).

  22. 22.

    , , , & Overexpression of Bcl-xS sensitizes MCF-7 cells to chemotherapy-induced apoptosis. Cancer Res. 55, 2507–2510 (1995).

  23. 23.

    , , , & Bcl-xS gene therapy induces apoptosis of human mammary tumors in nude mice. Cancer Res. 56 , 1965–1969 (1996).

  24. 24.

    & Mechanisms of multidrug resistance in cancer treatment. Acta Oncol. 31, 205 –213 (1992).

  25. 25.

    , & Bcl-xS antagonizes the protective effects of Bcl-xL. J. Biol. Chem. 271, 6306– 6312 (1996).

  26. 26.

    , & Bcl-xS and Bad potentiate the death suppressing activities of Bcl-xL, Bcl-2, and A1 in yeast. J. Biol. Chem. 273 , 23704–23708 (1998).

  27. 27.

    et al. Enforced expression of Bcl-XS induces differentiation and sensitizes chronic myelogenous leukemia-blast crisis K562 cells to 1-beta-D-arabinofuranosylcytosine-mediated differentiation and apoptosis. Cell Growth & Differ. 7, 1617–1623 (1996).

  28. 28.

    , & Bcl-x expression influences keratinocyte cell survival but not terminal differentiation. Cell Growth & Differ. 8, 619–629 (1997).

  29. 29.

    , , , & Expression of Bcl-XS alters cytokinetics and decreases clonogenic survival in K12 rat colon carcinoma cells. Oncogene 17, 2981–2991 (1998).

  30. 30.

    et al. Selective inhibition of mutant Ha-ras mRNA expression by antisense oligonucleotides. J. Biol. Chem. 267, 19954 –19962 (1992).

  31. 31.

    & Altered mRNA splicing and inhibition of E-selectin by an antisense oligonucleotide in human umbilical vein endothelial cells. J. Biol. Chem. 271, 30398– 30403 (1996).

  32. 32.

    , , , & TNF receptor death domain-associated proteins TRADD and FADD signal activation of acid sphingomyelinase. J. Biol. Chem. 273, 5916–5922 (1998).

Download references

Acknowledgements

We would like to thank Sue Freier, Todd Burckin, and Frank Bennett for advice with these studies.

Author information

Affiliations

  1. Department of Pharmacology, Carlsbad, CA 92008.

    • Jennifer K. Taylor
    •  & Nicholas M. Dean
  2. Department of Structural Biology, Isis Pharmaceuticals , Carlsbad, CA 92008.

    • Qing Qing Zhang
    •  & Jacqueline R. Wyatt

Authors

  1. Search for Jennifer K. Taylor in:

  2. Search for Qing Qing Zhang in:

  3. Search for Jacqueline R. Wyatt in:

  4. Search for Nicholas M. Dean in:

Corresponding author

Correspondence to Nicholas M. Dean.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/15079

Further reading