Tumor Biology

, Volume 37, Issue 5, pp 5919–5923 | Cite as

EZH2 mediates ATO-induced apoptosis in acute myeloid leukemia cell lines through the Wnt signaling pathway

  • Hao Zhang
  • Huizi Gu
  • Limei Li
  • Yuan Ren
  • Lijun Zhang
Original Article


In this study, we examined the mechanisms associated with EZH2 mediation of apoptosis and chemoresistance to arsenic trioxide (ATO) in acute myeloid leukemia (AML) cell lines through the Wnt/β-catenin signaling pathway. The induction of spontaneous apoptosis observed in multiple EZH2-silenced leukemic cell lines was assessed by flow cytometry, and levels of Wnt/β-catenin-related expression were determined by western blot analysis. In comparison with AML control cells, EZH2-knockdown cells exhibited increased apoptosis and significant downregulation of β-catenin expression, as well as decreases in GSK-3β phosphorylation and β-catenin activation (p < 0.05 for all measurements). Additionally, EZH2 knockdown sensitized AML cells to induced cell death following administration of chemotherapeutic ATO. Our results suggested that EZH2 in leukemic cell lines might inhibit ATO-induced apoptosis and that EZH2 may be a potential therapeutic target in AML patients undergoing ATO treatment. Our findings provide new insights into the role of ATO and EZH2 in regulating AML progression.


Acute myeloid leukemia ATO EZH2 Wnt/β-catenin 



This study was funded by the National Natural Science Foundation of China (No. 81402384) and the Natural Science Foundation of Liaoning Province (201202288).

Author contributions

Hao Zhang and Yuan Ren carried out the molecular genetic studies and drafted the manuscript. Limei Li and Lijun Zhang participated in the design of the study, and Huizi Gu performed the statistical analysis. Lijun Zhang conceived of the study, participated in its design and coordination, and helped draft the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflicts of interest



  1. 1.
    Estey EH. Acute myeloid leukemia: 2013 update on risk-stratification and management. Am J Hematol. 2013;88(4):318–27.CrossRefPubMedGoogle Scholar
  2. 2.
    Zhang T, Li Y. Clinical analysis and research on Ai-lin 1 treatment in acute myeloid leukemia. Chin J Integr Chin West Med. 1984;4:19–20.Google Scholar
  3. 3.
    Sun HD, Ma L, Hu XC, Zhang TD. Ai-Lin I treated 32 cases of acute promyelocytic leukemia. Chin J Integr Chin West Med. 1992;12:170–1.Google Scholar
  4. 4.
    Zhang P, Wang SY, Hu XH. Arsenic trioxide treated 72 cases of acute promyelocytic leukemia. Chin J Hematol (Chin). 1996;17:58–62.Google Scholar
  5. 5.
    Varambally S, Cao Q, Mani RS, et al. Genomic loss of microRNA-101 leads to overexpression of histone methyltransferase EZH2 in cancer. Science. 2008;322:1695–9.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Varambally S, Dhanasekaran SM, Zhou M, et al. The polycomb group protein EZH2 is involved in progression of prostate cancer. Nature. 2002;419:624–9.CrossRefPubMedGoogle Scholar
  7. 7.
    Kleer CG, Cao Q, Varambally S, et al. EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci U S A. 2003;100:11606–11.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Wagener N, Macher-Goeppinger S, Pritsch M, et al. Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma. BMC Cancer. 2010;10:524.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Takawa M, Masuda K, Kunizaki M, et al. Validation of the histonemethyltransferase EZH2 as a therapeutic target for various types of human cancer and as a prognostic marker. Cancer Sci. 2011;102:1298–305.CrossRefPubMedGoogle Scholar
  10. 10.
    Morin RD, Mendez-Lago M, Mungall AJ, et al. Frequent mutation of histone-modifying genes in non-Hodgkin lymphoma. Nature. 2011;476:298–303.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Morin RD, Johnson NA, Severson TM, et al. Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat Genet. 2010;42:181–5.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Pasqualucci L, Trifonov V, Fabbri G, et al. Analysis of the coding genome of diffuse large B-cell lymphoma. Nat Genet. 2011;43:830–7.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    McCabe MT, Graves AP, Ganji G, et al. Mutation of A677 in histonemethyltransferase EZH2in human B-cell lymphoma promotes hypertrimethylation of histone H3 on lysine 27 (H3K27). Proc Natl Acad Sci U S A. 2012;109:2989–94.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Ryan RJ, Nitta M, Borger D, et al. EZH2 codon 641 mutations are common in BCL2-rearranged germinal center B cell lymphomas. PLoS ONE. 2011;6, e28585.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell. 2009;17(1):9–26.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Murgo AJ. Clinical trials of arsenic trioxide in hematologic and solid tumors: overview of the National Cancer Institute Cooperative Research and Development Studies. Oncologist. 2001;6 Suppl 2:22–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Tallman MS. What is the role of arsenic in newly diagnosed APL? Best Pract Res Clin Haematol. 2008;21:659–66.CrossRefPubMedGoogle Scholar
  18. 18.
    Berenson JR, Yeh HS. Arsenic compounds in the treatment of multiple myeloma: a new role for a historical remedy. Clin Lymphoma Myeloma. 2006;7:192–8.CrossRefPubMedGoogle Scholar
  19. 19.
    Baj G, Arnulfo A, Deaglio S, et al. Arsenic trioxide and breast cancer: analysis of the apoptotic, differentiative and immunomodulatory effects. Breast Cancer Res Treat. 2002;73:61–73.CrossRefPubMedGoogle Scholar
  20. 20.
    Evens AM, Tallman MS, Gartenhaus RB. The potential of arsenic trioxide in the treatment of malignant disease: past, present, and future. Leuk Res. 2004;28:891–900.CrossRefPubMedGoogle Scholar
  21. 21.
    Chen Z, Chen GQ, Shen ZX, et al. Expanding the use of arsenic trioxide: leukemias and beyond. Semin Hematol. 2002;39:22–6.CrossRefPubMedGoogle Scholar
  22. 22.
    Dilda PJ, Hogg PJ. Arsenical-based cancer drugs. Cancer Treat Rev. 2007;33:542–64.CrossRefPubMedGoogle Scholar
  23. 23.
    Verstovsek S, Giles F, Quintas-Cardama A, et al. Arsenic derivatives in hematologic malignancies: a role beyond acute promyelocytic leukemia? Hematol Oncol. 2006;24:181–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Kühnl A, Valk PJ, Sanders MA, et al. Downregulation of the Wnt inhibitor CXXC5 predicts a better prognosis in acute myeloid leukemia. Blood. 2015;125(19):2985–94.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Giambra V, Jenkins CE, Lam SH, et al. Leukemia stem cells in T-ALL require active Hif1α and Wnt signaling. Blood. 2015;125(25):3917–27.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Wang Y, Krivtsov AV, Sinha AU, et al. The Wnt/beta-catenin pathway is required for the development of leukemia stem cells in AML. Science. 2010;327(5973):1650–3.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Hao Zhang
    • 1
    • 2
  • Huizi Gu
    • 3
  • Limei Li
    • 4
  • Yuan Ren
    • 1
  • Lijun Zhang
    • 1
  1. 1.Department of HematologyThe First Hospital of China Medical UniversityShenyangChina
  2. 2.Breast Disease and Reconstruction Center, Breast Cancer Key Lab of DalianThe Second Hospital of Dalian Medical UniversityDalianChina
  3. 3.Department of NeurologyThe Second Hospital of Dalian Medical UniversityDalianChina
  4. 4.Department of HematologyThe People’s Hospital of Liaoning ProvinceShenyangChina

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