Skip to main content

Advertisement

SpringerLink
Log in

Effect of siRNA targeting Ets2 gene on chemosensitization of human acute monocytic leukemic cell line SHI-1

  • Published:
The Chinese-German Journal of Clinical Oncology

Abstract

Objective

This study was to observe the levels of Ets2 mRNA expression in leukemia patients and investigate the effect of small interfering RNA (siRNA) targeting Ets2 gene on sensibility of human acute monocytic leukemic cell line SHI-1 cells to etoposide (VP-16). SHI-1 cells to etoposide (VP-16).

Methods

Ets2 mRNA levels were determined by reverse transcription polymerase chain reaction (RT-PCR). reaction (RT-PCR). After the transfection of Ets2 siRNA to SHI-1 cells by electroporation method, qRT-PCR was used to detect Ets2 gene expression in these cells; VP-16-induced apoptosis was investigated by Annexin V-FITC/PI. Ets2 gene expression in these cells; VP-16-induced apoptosis was investigated by Annexin V-FITC/PI.

Results

Ets2 mRNA was detectable in SHI-cells. was detectable in SHI-cells. The Ets2 expression rate was respectively 10% in 5 healthy volunteers, 60% in 5 acute lymphocytic leukemia (ALL) patients, 73. leukemia (ALL) patients, 73. 68% in 19 acute nonlymphocytic leukemia (ANLL) patients and 100% in 4 chronic myeloid leukemia (CML) patients. leukemia (CML) patients. The expression levels of Ets2 mRNA were significantly higher in leukemia patients compared with healthy volunteers. healthy volunteers. It also showed that siRNA targeting Ets2 gene resulted in substantial loss of Ets2 mRNA of SHI-1 cells compared to the control groups. compared to the control groups. Downregulation of Ets2 gene expression increased SHI-1 cells apoptosis and VP-16-induced apoptosis of SHI-1 cells. apoptosis of SHI-1 cells.

Conclusion

The high-level expression of Ets2 transcription factor in leukemia cells were connected with proliferation and anti-apoptosis of leukemia cells. with proliferation and anti-apoptosis of leukemia cells. SiRNA mediated Ets2 gene silencing induced cell apoptosis and enhanced in vitro sensitivity to chemotherapy (VP-16) of SHI-1 cells. in vitro sensitivity to chemotherapy (VP-16) of SHI-1 cells. It speculated the high-level expression of Ets2 may actually be an unfavorable determinant of chemotherapy sensitivity in leukemia. be an unfavorable determinant of chemotherapy sensitivity in leukemia.

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

References

  1. Yordy JS, Muise-Helmericks RC. Signal transduction and the Ets family of transcription factors. Oncogene, 2000, 19: 6503–6513.

    Article  PubMed  CAS  Google Scholar 

  2. de Nigris F, Mega T, Berger N, et al. Induction of ETS1 and ETS2 transcription factors is required for thyroid cell transformation. Cancer Res, 2001, 61: 2267–2275.

    PubMed  Google Scholar 

  3. Li X, Lu JY, Zhao LQ, et al. Overexpression of ETS2 in human esophageal squamous cell carcinoma. World J Gastroenterol, 2003, 9: 205–208.

    PubMed  Google Scholar 

  4. Carbone GM, Napoli S, Valentini A, et al. Triplex DNA-mediated downregulation of Ets2 expression results in growth inhibition and apoptosis in human prostate cancer cells. Nucleic Acids Res, 2004, 32: 4358–4367.

    Article  PubMed  CAS  Google Scholar 

  5. Foos G, Hauser CA. Altered Ets transcription factor activity in prostate tumor cells inhibits anchorage-independent growth, survival and invasiveness. Oncogene, 2000, 19: 5507–5516.

    Article  PubMed  CAS  Google Scholar 

  6. Buggy Y, Maguire TM, McDermott E, et al. Ets2 transcription factor in normal and neoplastic human breast tissue. Eur J Cancer, 2006, 42: 485–491.

    Article  PubMed  CAS  Google Scholar 

  7. Zabuawala T, Taffany DA, Sharma SM, et al. An ets2-driven transcriptional program in tumor-associated macrophages promotes tumor metastasis. Cancer, Res, 2010, 70: 1323–1333.

    Article  CAS  Google Scholar 

  8. Dwyer JM, Liu JP. Ets2 transcription factor, telomerase activity and breast cancer. Clin Exp Pharmacol Physiol, 2010, 37: 83–87.

    Article  PubMed  CAS  Google Scholar 

  9. Santoro A, Maggio A, Carbone P, et al. Amplification of ETS2 oncogene in acute nonlymphoblastic leukemia with t (6;21;18). Cancer Genet Cytogenet, 1992, 58: 71–75.

    Article  PubMed  CAS  Google Scholar 

  10. Ge Y, LaFiura KM, Dombkowski AA, et al. The role of the proto-oncogene ETS2 in acute megakaryocytic leukemia biology and therapy. Leukemia, 2008, 22: 521–529.

    Article  PubMed  CAS  Google Scholar 

  11. Zeller B, Gustafsson G, Forestier E, et al. Acute leukaemia in children with Down syndrome: a population-based Nordic study. Br J Haematol, 2005, 128: 797–804.

    Article  PubMed  Google Scholar 

  12. Stankiewicz MJ, Crispino JD. ETS2 and ERG promote megakaryopoiesis and synergize with alterations in GATA-1 to immortalize hematopoietic progenitor cells. Blood, 2009, 113: 3337–3347.

    Article  PubMed  CAS  Google Scholar 

  13. Baldus CD, Liyanarachchi S, Mrózek K, et al. Acute myeloid leukemia with complex karyotypes and abnormal chromosome 21: Amplification discloses overexpression of APP, ETS2, and ERG genes. Proc Natl Acad Sci U S A, 2004, 101: 3915–3920.

    Article  PubMed  CAS  Google Scholar 

  14. Elbashir SM, Harborth J, Lendeckel W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 2001, 411: 494–498.

    Article  PubMed  CAS  Google Scholar 

  15. Wiese M, Castiglione K, Hensel M, et al. Small interfering RNA (siRNA) delivery into murine bone marrow-derived macrophages by electroporation. J Immunol Methods, 2010, 353: 102–110.

    Article  PubMed  CAS  Google Scholar 

  16. Chen S, Xue Y, Zhang X, et al. A new human acute monocytic leukemia cell line SHI-1 with t(6;11)(q27;q23), p53 gene alterations and high tumorigenicity in nude mice. Haematologica, 2005, 90: 766–775.

    PubMed  CAS  Google Scholar 

  17. Tomonaga M. New classification of leukemia. Rinsho Byori, 2001 Suppl 15, 3: 45–53.

    Google Scholar 

  18. Maroulakou IG, Bowe DB. Expression and function of Ets transcription factors in mammalian development: a regulatory network. Oncogene, 2000, 19: 6432–6442.

    Article  PubMed  CAS  Google Scholar 

  19. Jin DI, Jameson SB, Reddy MA, et al. Alterations in differentiation and behavior of monocytic phagocytes in transgenic mice that express dominant suppressors of ras signaling. Mol Cell Biol, 1995, 15: 693–703.

    PubMed  CAS  Google Scholar 

  20. Boulukos KE, Pognonec P, Sariban E, et al. Rapid and transient expression of Ets2 in mature macrophages following stimulation with cMGF, LPS, and PKC activators. Genes Dev, 1990, 4: 401–409.

    Article  PubMed  CAS  Google Scholar 

  21. Sevilla L, Zaldumbide A, Carlotti F, et al. Bcl-XL expression correlates with primary macrophage differentiation, activation of functional competence, and survival and results from synergistic transcriptional activation by Ets2 and PU.1. J Biol Chem, 2001, 276: 17800–17807.

    Article  PubMed  CAS  Google Scholar 

  22. Sevilla L, Aperlo C, Dulic V, et al. The Ets2 transcription factor Inhibits apoptosis induced by colony-stimulating factor-1 deprivation of macrophages through a Bcl-xL -dependent mechanism. Mol Cell Biol, 1999, 19: 2624–2634.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Chun Huang, Lifang Wang, Chunrui Li, Shan Huang & Dengju Li

Authors
  1. Chun Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lifang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chunrui Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shan Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dengju Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dengju Li.

Additional information

Supported by a grant from the Natural Science Foundation of Hubei Province (No. 2009CDB416).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, C., Wang, L., Li, C. et al. Effect of siRNA targeting Ets2 gene on chemosensitization of human acute monocytic leukemic cell line SHI-1. Chin. -Ger. J. Clin. Oncol. 10, 726–729 (2011). https://doi.org/10.1007/s10330-011-0864-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10330-011-0864-x

Key words

Search

Navigation