Skip to main content
SpringerLink
Log in

Downregulation of activated leukemic oncogenes AML1-ETO and RUNX1(K83N) expression with RNA-interference

  • Cell Molecular Biology
  • Published:
Molecular Biology Aims and scope Submit manuscript

Abstract

In the present study, we have applied the siRNA approach to reduce the expression of AML1-ETO and RUNX1(K83N) oncogenes, which are frequently found in leukemic cells. We have designed small hairpin RNAs (shRNA) for targeting AML1-ETO oncogene and a region close to the 5′-untranslated region of mRNA for the mutant RUNX1(K83N) oncogene and expressed the shRNAs in lentiviral vectors. We report a stable reduction in expression of oncogenes following the introduction of shRNAs into cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

AML:

Acute Myeloid Leykosis

FAB:

Franco-American-British classification of leucosis

HSC:

Hemopoietic Stem Cell

siRNA:

small interfering RNA

shPHK:

small hairpin interfering RNA.

References

  1. Weinberg R.A. 2007. The Biology of Cancer. NY: Garland Science.

    Google Scholar 

  2. Jemal A., Siegel R., Ward E., Hao Y., Xu J., Thun M.J. 2009. Cancer statistics. Ca. Cancer. J. Clin. 59, 225–249.

    Article  PubMed  Google Scholar 

  3. Peterson L.F., Zhang D.E. 2004. The 8;21 translocation in leukemogenesis. Oncogene. 23, 4255–4562.

    Article  CAS  PubMed  Google Scholar 

  4. Imai Y., Kurokawa M., Izutsu K., et al. 2000. Mutations of the AML1 gene in myelodysplastic syndrome and their functional implications in leukemogenesis. Blood. 96, 3154–3160.

    CAS  PubMed  Google Scholar 

  5. Osato M., Asou N., Abdalla E., et al. 1999. Biallelic and heterozygous point mutations in the runt domain of the AML1/PEBP2B gene associated with myeloblastic leukemia. Blood. 93, 1817–1824.

    CAS  PubMed  Google Scholar 

  6. Michaud J., Wu F., Osato M., et al. 2002. In vitro analyses of known and novel RUNX1/AML1 mutations in dominant familial platelet disorder with predisposition to acute myelogenous leukemia: implications for mechanisms of pathogenesis. Blood. 99, 1364–1372.

    Article  CAS  PubMed  Google Scholar 

  7. Cammenga J., Niebuhr B., Horn S., Bergholz U., Putz G., Buchholz F., Löhler J., Stocking C. 2007. RUNX1 DNA-binding mutants, associated with minimally differentiated acute myelogenous leukemia, disrupt myeloid differentiation. Cancer Res. 67, 537–545.

    Article  CAS  PubMed  Google Scholar 

  8. Wang Y.Y., Zhou G.B., Yin T., et al. 2005. AML1-ETO and C-KIT mutation/overexpression in t(8;21) leukemia: Implication in stepwise leukemogenesis and response to Gleevec. Proc. Natl. Acad. Sci. USA. 102, 1104–1109.

    Article  CAS  PubMed  Google Scholar 

  9. Peterson L.F., Boyapati A., Ahn E.Y., Biggs J.R., Okumura A.J., Lo M.C., Yan M., Zhang D.E. 2007. Acute myeloid leukemia with the 8q22;21q22 translocation: Secondary mutational events and alternative t(8;21) transcripts. Blood. 110, 799–805.

    Article  CAS  PubMed  Google Scholar 

  10. Bantounas I., Phylactou L.A., Uney J.B. 2004. RNA interference and the use of small interfering RNA to study gene function in mammalian systems. J. Mol. Endocrinol. 33, 545–557.

    Article  CAS  PubMed  Google Scholar 

  11. Vilgelm A.E., Chumakov S.P., Prassolov V.S. 2006. RNA interference: Biology and prospects of application in biomedicine and biotechnology. Mol. Biol. 40, 339–354.

    Article  CAS  Google Scholar 

  12. Elbashir S.M., Harborth J., Lendeckel W., Yalcin A., Weber K., Tuschl T. 2001. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411, 494–498.

    Article  CAS  PubMed  Google Scholar 

  13. Caplen N.J., Parrish S., Imani F., Fire A., Morgan R.A. 2001. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc. Natl. Acad. Sci. USA. 98, 9742–9747.

    Article  CAS  PubMed  Google Scholar 

  14. Fang Z.H., Dong C.L., Chen Z., Zhou B., Liu N., Lan H.F., Liang L., Liao W.B., Zhang L., Han Z.C. 2009. Transcriptional regulation of survivin by c-Myc in BCR/ABL-transformed cells: Implications in anti-leukaemic strategy. J. Cell Mol. Med. 13, 2039–2052.

    Article  PubMed  Google Scholar 

  15. Heidenreich O. 2009. Targeting oncogenes with siRNAs. Methods Mol. Biol. 487, 221–242.

    CAS  PubMed  Google Scholar 

  16. Thomas M., Greil J., Heidenreich O. 2006. Targeting leukemic fusion proteins with small interfering RNAs: Recent advances and therapeutic potentials. Acta Pharmacol. Sin. Mar. 27, 273–281.

    Article  CAS  Google Scholar 

  17. Nechipurenko O., Rossi L., Moore B. 2007. Comparison of approaches for rational siRNA design leading to a new efficient and transparent method. Nucleic Acids Res. 35, 1–10.

    Google Scholar 

  18. Volkov A.A., Kruglova N.S., Meschaninova M.I., Venyaminova A.G., Zenkova M.A., Vlassov V.V., Chernolovskaya E.L. 2009. Selective protection of nuclease-sensitive sites in siRNA prolongs silencing effect. Oligonucleotides. 19, 191–202.

    Article  CAS  PubMed  Google Scholar 

  19. Fagard M., Vaucheret H. 2000. Systemic silencing signals. Plant Mol. Biol. 43, 285–293.

    Article  CAS  PubMed  Google Scholar 

  20. Singer O., Verma I.M. 2008. Applications of lentiviral vectors for shRNA delivery and transgenesis. Curr. Gene. Ther. 8, 483–488.

    Article  CAS  PubMed  Google Scholar 

  21. Manjunath N., Wu H., Subramanya S., Shankar P. 2009. Lentiviral delivery of short hairpin RNAs. Adv. Drug Delivery Rev. 61, 732–745.

    Article  CAS  Google Scholar 

  22. Schwieger M., Löhler J., Friel J., Scheller M., Horak I., Stocking C. 2002. AML1-ETO inhibits maturation of multiple lymphohematopoietic lineages and induces myeloblast transformation in synergy with ICSBP deficiency. J. Exp. Med. 196, 1227–1240.

    Article  CAS  PubMed  Google Scholar 

  23. Kravchenko J.E., Ilyinskaya G.V., Komarov P.G., Agapova L.S., Kochetkov D.V., Strom E., Frolova E.I., Kovriga I., Gudkov A.V., Feinstein E., Chumakov P.M. 2008. Small-molecule RETRA suppresses mutant p53-bearing cancer cells through a p73-dependent salvage pathway. Proc. Natl. Acad. Sci. USA. 105, 6302–6307.

    Article  CAS  PubMed  Google Scholar 

  24. Bergemann J., Kuhlcke K., Fehse B., Ratz I., Ostertag W., Lother H. 1995. Excision of specific DNA-sequences from integrated retroviral vectors via site-specific recombination. Nucleic Acids Res. 21, 4451–4456.

    Article  Google Scholar 

  25. Spirin P.V., Baskaran D., Rubtsov P.M., Zenkova M.A., Vlasov V.V., Chernolovskaya E.L., Prassolov V. S. 2009. A comparison of target gene silencing using syntheti-cally modified siRNA and shRNA that express recombinant lentiviral vectors. Acta Naturae. 2, 86–90.

    Google Scholar 

  26. Takenokuchi M., Yashida C., Takeuchi K., Nakamachi Y., Mukai M., Kondo S., Kumagai S., Saigo K., Murayama T., Koizumi T., Tatsumi E. 2004. Quantitative nested reverse transcriptase PCR vs. real-time PCR for measuring AML1/ETO (MTG8) transcripts. Clin. Lab. Haem. 26, 107–114

    Article  CAS  Google Scholar 

  27. Koldehoff M., Kordelas L., Beelen D.W., Elmaagacli A.H. 2010. Small interfering RNA against BCR-ABL transcripts sensitize mutated T315I cells to nilotinib. Haematologica. 95, 388–397.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia

    P. V. Spirin, D. Baskaran, N. N. Orlova, A. V. Rulina, N. A. Nikitenko, P. M. Rubtsov, P. M. Chumakov & V. S. Prassolov

  2. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, 630090, Russia

    E. L. Chernolovskaya, M. A. Zenkova & V. V. Vlassov

  3. Heinrich-Pette-Institute for Experimental Virology and Immunology, Hamburg, D-20251, Germany

    C. Stocking

Authors
  1. P. V. Spirin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. Baskaran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N. N. Orlova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. V. Rulina
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. A. Nikitenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. E. L. Chernolovskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M. A. Zenkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. V. V. Vlassov
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. P. M. Rubtsov
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. P. M. Chumakov
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. C. Stocking
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. V. S. Prassolov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. S. Prassolov.

Additional information

Original Russian Text © P.V. Spirin, D. Baskaran, N.N. Orlova, A.V. Rulina, N.A. Nikitenko, E.L. Chernolovskaya, M.A. Zenkova, V.V. Vlassov, P.M. Rubtsov, P.M. Chumakov, C. Stocking, V.S. Prassolov, 2010, published in Molekulyarnaya Biologiya, 2010, Vol. 44, No. 5, pp. 876–888.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Spirin, P.V., Baskaran, D., Orlova, N.N. et al. Downregulation of activated leukemic oncogenes AML1-ETO and RUNX1(K83N) expression with RNA-interference. Mol Biol 44, 776–786 (2010). https://doi.org/10.1134/S0026893310050146

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893310050146

Key words

Search

Navigation