Skip to main content
SpringerLink
Log in

Modulation of activated oncogene c-kit expression with RNA-interference

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

Abstract

Overexpression of oncogene c-kit is detected in 80% patients with acute myeloid leukemia (AML). A transgenic model cell line expressing oncogene c-kit was obtained by transduction with a recombinant retrovirus. We have designed small interfering RNAs (siRNAs) that efficiently suppress the expression of activated oncogene c-kit. Further, small hairpin RNAs (shRNAs) targeting c-kit mRNA were designed and expressed in lentiviral vectors. We report a stable reduction in c-kit expression following the introduction of shRNAs into model cells, as well as Kasumi-1 cells from a patient with AML.

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 leukemia

HSC:

hemopoietic stem cell

FAB:

French-American-British classification system of leukemias

siRNA:

small interfering RNA

shRNA:

small hairpin RNA

References

  1. Weinberg R.A. 2007. The Biology of Cancer. N.Y.: Garland Sci.

    Google Scholar 

  2. 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  PubMed  CAS  Google Scholar 

  3. 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 

  4. Grimwade D., Walker H., Oliver F., Wheatley K., Harrison C., Harrison G., Rees J., Hann I., Stevens R., Burnett A., Goldstone A. 1998. The importance of diagnostic cytogenetics on outcome in AML: Analysis of 1,612 patients entered into the MRC AML 10 trial. The medical research council adult and children’s leukaemia working parties. Blood. 92, 2322–2333.

    PubMed  CAS  Google Scholar 

  5. Bennett J.M., Catovsky D., Daniel M.T., Flandrin G., Galton D.A., Gralnick H.R., Sultan C. 1976. Proposals for the classification of the acute leukaemias. French-American-British (FAB) co-operative group. Br. J. Haematol. 33, 451–458.

    Article  PubMed  CAS  Google Scholar 

  6. Gilliland D.G. 1998. Molecular genetics of human leukemia. Leukemia. 12(Suppl. 1), S7–S12.

    PubMed  CAS  Google Scholar 

  7. Frankfurt O., Licht J.D., Tallman M.S. 2007. Molecular characterization of acute myeloid leukemia and its impact on treatment. Curr. Opin. Oncol. 19, 635–649.

    Article  PubMed  Google Scholar 

  8. Ikeda H., Kanakura Y., Tamaki T., Kuriu A., Kitayama H., Ishikawa J., Kanayama Y., Yonezawa T., Tarui S., Griffin J.D. 1991. Expression and functional role of the proto-oncogene c-kit in acute myeloblastic leukemia cells. Blood. 78, 2962–2968.

    PubMed  CAS  Google Scholar 

  9. Baskaran D., Spirin P.V., Prassolov V.S. 2010. Activated leukemic oncogenes responsible for neoplastic transformation of hematopoietic cells. Mol. Biol. (Moscow). 44, 369–379.

    Article  CAS  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  PubMed  CAS  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. (Moscow). 40, 339–354.

    Article  CAS  Google Scholar 

  12. Spirin P.V., Baskaran D., Orlova N.N., Rulina A.V., Nikitenko N.A., Chernolovskaya E.L., Zenkova M.A., Vlassov V.V., Rubtsov P.M., Chumakov P.M., Stocking C., Prassolov V.S. 2010. Downregulation of activated leukemic oncogenes AML1-ETO and RUNX1(K83N) expression with RNA-interference. Mol. Biol. (Moscow). 44, 776–786.

    Article  CAS  Google Scholar 

  13. 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  PubMed  CAS  Google Scholar 

  14. 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. U.S.A. 98, 9742–9747.

    Article  PubMed  CAS  Google Scholar 

  15. 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 

  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.

    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  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  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  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  22. Larizza L., Magnani I., Beghini A. 2005. The Kasumi-1 cell line: A t(8;21)-kit mutant model for acute myeloid leukemia. Leuk. Lymphoma. 46, 247–255.

    Article  PubMed  CAS  Google Scholar 

  23. 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  PubMed  CAS  Google Scholar 

  24. 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. U.S.A. 105, 6302–6307.

    Article  PubMed  CAS  Google Scholar 

  25. Huo L., Sugimura J., Tretiakova M.S., Patton K.T., Gupta R., Popov B., Laskin W.B., Yeldandi A., Teh B.T., Yang X.J. 2005. C-kit expression in renal oncocytomas and chromophobe renal cell carcinomas. Hum. Pathol. 36, 262–268.

    Article  PubMed  CAS  Google Scholar 

  26. Stec R., Grala B., Maczewski M., Bodnar L., Szczylik C. 2009. Chromophobe renal cell cancer: Review of the literature and potential methods of treating metastatic disease. J. Exp. Clin. Cancer Res. 28, 1–6.

    Article  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, N. A. Nikitenko, T. D. Lebedev, P. M. Rubtsov & V. S. Prasolov

  2. 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. N. A. Nikitenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. D. Lebedev
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. M. Rubtsov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. Stocking
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. V. S. Prasolov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. S. Prasolov.

Additional information

Original Russian Text © P.V. Spirin, N.A. Nikitenko, T.D. Lebedev, P.M. Rubtsov, C. Stocking, V.S. Prasolov, 2011, published in Molekulyarnaya Biologiya, 2011, Vol. 45, No. 6, pp. 1036–1045.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Spirin, P.V., Nikitenko, N.A., Lebedev, T.D. et al. Modulation of activated oncogene c-kit expression with RNA-interference. Mol Biol 45, 950–958 (2011). https://doi.org/10.1134/S0026893311060136

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893311060136

Keywords

Search

Navigation