Tumor Biology

, Volume 35, Issue 4, pp 3933–3940 | Cite as

Knockdown of CD44 enhances chemosensitivity of acute myeloid leukemia cells to ADM and Ara-C

  • Ni-sha Wang
  • Min Wei
  • Wen-li Ma
  • Wei Meng
  • Wen-ling Zheng
Research Article


It is known that chemoresistance is a major cause of treatment failure in acute myeloid leukemia (AML). Substantial data indicate that the CD44 adhesion molecule is strongly expressed on AML blasts and that it can also inhibit apoptosis. Our study shows that drug resistance of the AML cell line HL60/ADM is due to overexpression of CD44. In an in vitro study, we knocked down CD44 in the HL60/ADM cell line using small interfering RNA (siRNA). Cell proliferation and the 50 % inhibitory concentrations (IC50) were determined by Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis and intracellular ADM accumulation were detected by flow cytometry. Expression of CD44, Bcl-2, c-Myc were assayed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The results indicate that the expression of CD44 in HL60/ADM cell line was much higher than in HL60 cell, and siRNA targeted CD44 (siRNA/CD44) could silence its expression in both mRNA and protein levels effectively. siRNA/CD44 substantially induces cell apoptosis, inhibits cell proliferation, enhances susceptibility to ADM and Ara-C, and at the same time increases intracellular ADM accumulation even reverses chemoresistance to ADM and Ara-C. Furthermore, by qRT-PCR and Western blot, we found that siRNA/CD44 decreases Bcl-2 and c-Myc synthesis. Our study provides a novel clue that CD44 plays a significant role in the chemoresistance of AML cells to Ara-C and ADM. Moreover, this provides a new direction to the approaches that combination therapy including targeting CD44 may overcome drug resistance and improve treatment effects.


CD44 Apoptosis Chemosensitivity Ara-C (cytosine arabinoside) ADM (adriamycin) 



This work is supported by the National Natural Science Foundation of China (Grant No. 30901721) and the Natural science foundation of Guangdong province (No: S2012010009537).

Conflicts of interest



  1. 1.
    Shipley JL, Butera JN. Acute myelogenous leukemia. Exp Hematol. 2009;37(6):649–58.PubMedCrossRefGoogle Scholar
  2. 2.
    Tallman MS, Gilliland DG, Rowe JM. Drug therapy for acute myeloid leukemia. Blood. 2005;106(4):1154–63.PubMedCrossRefGoogle Scholar
  3. 3.
    Hiddemann W. Cytosine arabinoside in the treatment of acute myeloid leukemia: the role and place of high-dose regimens. Ann Hematol. 1991;62(4):119–28.PubMedCrossRefGoogle Scholar
  4. 4.
    Konopleva MY, Jordan CT. Leukemia stem cells and microenvironment: biology and therapeutic targeting. J Clin Oncol. 2011;29(5):591–9.PubMedCrossRefGoogle Scholar
  5. 5.
    Ayala F, Dewar R, Kieran M, Kalluri R. Contribution of bone microenvironment to leukemogenesis and leukemia progression. Leukemia. 2009;23(12):2233–41.PubMedCrossRefGoogle Scholar
  6. 6.
    Westhoff MA, Fulda S. Adhesion-mediated apoptosis resistance in cancer. Drug Resist Updat. 2009;12(4–5):127–36.PubMedCrossRefGoogle Scholar
  7. 7.
    Legras S, Gunthert U, Stauder R, Curt F, Oliferenko S, Kluin-Nelemans HC, et al. A strong expression of CD44-6v correlates with shorter survival of patients with acute myeloid leukemia. Blood. 1998;91(9):3401–13.PubMedGoogle Scholar
  8. 8.
    Charrad RS, Gadhoum Z, Qi J, Glachant A, Allouche M, Jasmin C, et al. Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood. 2002;99(1):290–9.PubMedCrossRefGoogle Scholar
  9. 9.
    Allouche M, Charrad RS, Bettaieb A, Greenland C, Grignon C, Smadja-Joffe F. Ligation of the CD44 adhesion molecule inhibits drug-induced apoptosis in human myeloid leukemia cells. Blood. 2000;96(3):1187–90.PubMedGoogle Scholar
  10. 10.
    Tamada M, Nagano O, Tateyama S, Ohmura M, Yae T, Ishimoto T, et al. Modulation of glucose metabolism by CD44 contributes to antioxidant status and drug resistance in cancer cells. Cancer Res. 2012;72(6):1438–48.PubMedCrossRefGoogle Scholar
  11. 11.
    Hu Z, Gao J, Zhang D, Liu Q, Yan L, Gao L, et al. High expression of Lewis y antigen and CD44 is correlated with resistance to chemotherapy in epithelial ovarian cancers. PLoS One. 2013;8(2):e57250.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Yan S, Ma D, Ji M, Guo D, Dai J, Zhao P, et al. Expression profile of Notch-related genes in multidrug resistant K562/A02 cells compared with parental K562 cells. Int J Lab Hematol. 2010;32(2):150–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Tothova E, Fricova M, Stecova N, Kafkova A, Elbertova A. High expression of Bcl-2 protein in acute myeloid leukemia cells is associated with poor response to chemotherapy. Neoplasma. 2002;49(3):141–4.PubMedGoogle Scholar
  14. 14.
    Konopleva M, Zhao S, Hu W, Jiang S, Snell V, Weidner D, et al. The anti-apoptotic genes Bcl-X(L) and Bcl-2 are over-expressed and contribute to chemoresistance of non-proliferating leukaemic CD34+ cells. Br J Haematol. 2002;118(2):521–34.PubMedCrossRefGoogle Scholar
  15. 15.
    Hoffman B, Amanullah A, Shafarenko M, Liebermann DA. The proto-oncogene c-myc in hematopoietic development and leukemogenesis. Oncogene. 2002;21(21):3414–21.PubMedCrossRefGoogle Scholar
  16. 16.
    Tsuruo T, Naito M, Tomida A, Fujita N, Mashima T, Sakamoto H, et al. Molecular targeting therapy of cancer: drug resistance, apoptosis and survival signal. Cancer Sci. 2003;94(1):15–21.PubMedCrossRefGoogle Scholar
  17. 17.
    Mahadevan D, List AF. Targeting the multidrug resistance-1 transporter in AML: molecular regulation and therapeutic strategies. Blood. 2004;104(7):1940–51.PubMedCrossRefGoogle Scholar
  18. 18.
    Sansonetti A, Bourcier S, Durand L, Chomienne C, Smadja-Joffe F, Robert-Lezenes J. CD44 activation enhances acute monoblastic leukemia cell survival via Mcl-1 upregulation. Leuk Res. 2012;36(3):358–62.PubMedCrossRefGoogle Scholar
  19. 19.
    Quere R, Andradottir S, Brun AC, Zubarev RA, Karlsson G, Olsson K, et al. High levels of the adhesion molecule CD44 on leukemic cells generate acute myeloid leukemia relapse after withdrawal of the initial transforming event. Leukemia. 2011;25(3):515–26.PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Bourguignon LY, Peyrollier K, Xia W, Gilad E. Hyaluronan-CD44 interaction activates stem cell marker Nanog, Stat-3-mediated MDR1 gene expression, and ankyrin-regulated multidrug efflux in breast and ovarian tumor cells. J Biol Chem. 2008;283(25):17635–51.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Bjorklund CC, Baladandayuthapani V, Lin HY, Jones RJ, Kuiatse I, Wang H, et al. Evidence of a role for CD44 and cell adhesion in mediating resistance to lenalidomide in multiple myeloma: therapeutic implications. Leukemia. 2013.Google Scholar
  22. 22.
    Testa U, Riccioni R. Deregulation of apoptosis in acute myeloid leukemia. Haematologica. 2007;92(1):81–94.PubMedCrossRefGoogle Scholar
  23. 23.
    Brunelle JK, Ryan J, Yecies D, Opferman JT, Letai A. MCL-1-dependent leukemia cells are more sensitive to chemotherapy than BCL-2-dependent counterparts. J Cell Biol. 2009;187(3):429–42.PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Park YS, Huh JW, Lee JH, Kim HR. shRNA against CD44 inhibits cell proliferation, invasion and migration, and promotes apoptosis of colon carcinoma cells. Oncol Rep. 2012;27(2):339–46.PubMedGoogle Scholar
  25. 25.
    Huang MJ, Cheng YC, Liu CR, Lin S, Liu HE. A small-molecule c-Myc inhibitor, 10058-F4, induces cell-cycle arrest, apoptosis, and myeloid differentiation of human acute myeloid leukemia. Exp Hematol. 2006;34(11):1480–9.PubMedCrossRefGoogle Scholar
  26. 26.
    Cheng YC, Lin H, Huang MJ, Chow JM, Lin S, Liu HE. Downregulation of c-Myc is critical for valproic acid-induced growth arrest and myeloid differentiation of acute myeloid leukemia. Leuk Res. 2007;31(10):1403–11.PubMedCrossRefGoogle Scholar
  27. 27.
    Liu L, Wang S, Chen R, Wu Y, Zhang B, Huang S, et al. Myc induced miR-144/451 contributes to the acquired imatinib resistance in chronic myelogenous leukemia cell K562. Biochem Biophys Res Commun. 2012;425(2):368–73.PubMedCrossRefGoogle Scholar
  28. 28.
    Song G, Liao X, Zhou L, Wu L, Feng Y, Han ZC. HI44a, an anti-CD44 monoclonal antibody, induces differentiation and apoptosis of human acute myeloid leukemia cells. Leuk Res. 2004;28(10):1089–96.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  1. 1.Institute of Genetic Engineering of Southern Medical UniversityGuangzhouChina

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