siRNA-mediated Silencing of Survivin Inhibits Proliferation and Enhances Etoposide Chemosensitivity in Acute Myeloid Leukemia Cells

  • Karami, Hadi (Immunology Research Center, Tabriz University of Medical Sciences) ;
  • Baradaran, Behzad (Immunology Research Center, Tabriz University of Medical Sciences) ;
  • Esfahani, Ali (Hematology and Oncology Research Center, Tabriz University of Medical Sciences) ;
  • Estiar, Mehrdad Asghari (Scientific Research Center, Tehran University of Medical Sciences) ;
  • Naghavi-Behzad, Mohammad (Medical Philosophy and History Research, Tabriz University of Medical Sciences) ;
  • Sakhinia, Masoud (Faculty of Medicine, University of Liverpool) ;
  • Sakhinia, Ebrahim (Tuberculosis and Lung Diseases Research Center, Tabriz University of Medical Sciences)
  • Published : 2013.12.31


Background: Overexpression of survivin, a known inhibitor of apoptosis, is associated with tumor progression and drug resistance in numerous malignancies, including leukemias. The aim of this study was to investigate the effect of a specific survivin small interference RNA (siRNA) on proliferation and the sensitivity of HL-60 acute myeloid leukemia (AML) cells to the chemotherapeutic drug etoposide. Materials and Methods: The cells were transfected with siRNAs using Lipofectamine $^{TM}2000$ transfection reagent. Relative survivin mRNA and protein levels were measured by quantitative real-time PCR and Western blotting, respectively. Trypan blue exclusion assays were performed to monitor tumor cell proliferation after siRNA transfection. The cytotoxic effects of etoposide and survivin siRNA, alone and in combination, on leukemic cells were determined using MTT assay. Apoptosis was assessed by ELISA cell death assay. Results: Survivin siRNA markedly reduced both mRNA and protein expression levels in a time-dependent manner, leading to distinct inhibition of cell proliferation and increased spontaneous apoptosis. Surprisingly, survivin siRNA synergistically increased the cell toxic effects of etoposide. Moreover, survivin down-regulation significantly enhanced its induction of apoptosis. Conclusions: Our study suggests that down-regulation of survivin by siRNA can trigger apoptosis and overcome drug resistance of leukemia cells. Therefore, survivin siRNA may be an effective adjuvant in AML chemotherapy.


  1. Adida C, Recher C, Raffoux E, et al (2000). Expression and prognostic significance of survivin in de novo acute myeloid leukaemia. Br J Haematol, 111, 196-203.
  2. Aoki Y, Cioca DP, Oidaira H, Kamiya J, Kiyosawa K (2003). RNA interference may be more potent than antisense RNA in human cancer cell lines. Clin Exp Pharmacol Physiol, 30, 96-102.
  3. Baran Y, Gur B, Kaya P, et al (2007). Upregulation of multi drug resistance genes in doxorubicin resistant human acute myelogeneous leukemia cells and reversal of the resistance. Hematol, 12, 511-7.
  4. Brantl S (2002). Antisense-RNA regulation and RNA interference. Biochim Biophys Acta, 1575, 15-25.
  5. Chou TC, Talalay P (1984). Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul, 22, 27-55.
  6. Church DN, Talbot DC (2012). Survivin in solid tumors: rationale for development of inhibitors. Curr Oncol Rep, 14, 120-8.
  7. Coumar MS, Tsai FY, Kanwar JR, Sarvagalla S, Cheung CH (2013). Treat cancers by targeting survivin: just a dream or future reality? Cancer Treat Rev, 39, 802-11.
  8. Dai Y, Grant S (2007). Targeting multiple arms of the apoptotic regulatory machinery. Cancer Res, 67, 2908-11.
  9. Devi GR (2006). siRNA-based approaches in cancer therapy. Cancer Gene Ther, 13, 819-29.
  10. Feng W, Yoshida A, Ueda T (2013). YM155 induces caspase-8 dependent apoptosis through downregulation of survivin and Mcl-1 in human leukemia cells. Biochem Biophys Res Commun, 435, 52-7.
  11. Fukuda S, Pelus LM (2006). Survivin, a cancer target with an emerging role in normal adult tissues. Mol Cancer Ther, 5, 1087-98.
  12. Gautrey HL, Tyson-Capper AJ (2012). Regulation of Mcl-1 by SRSF1 and SRSF5 in cancer cells. PLoS One, 7, 51497.
  13. High LM, Szymanska B, Wilczynska-Kalak U, et al (2010). The Bcl-2 homology domain 3 mimetic ABT-737 targets the apoptotic machinery in acute lymphoblastic leukemia resulting in synergistic in vitro and in vivo interactions with established drugs. Mol Pharmacol, 77, 483-94.
  14. Ho MM, Hogge DE, Ling V (2008). MDR1 and BCRP1 expression in leukemic progenitors correlates with chemotherapy response in acute myeloid leukemia. Exp Hematol, 36, 433-42.
  15. Kang MH, Reynolds CP (2009). Bcl-2 inhibitors: targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res, 15, 1126-32.
  16. Kelly RJ, Lopez-Chavez A, Citrin D, Janik JE, Morris JC (2011). Impacting tumor cell-fate by targeting the inhibitor of apoptosis protein survivin. Mol Cancer, 10, 35.
  17. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method. Methods, 25, 402-8.
  18. Martins LM, Mesner PW, Kottke TJ, et al (1997). Comparison of caspase activation and subcellular localization in HL-60 and K562 cells undergoing etoposide-induced apoptosis. Blood, 90, 4283-96.
  19. Miao GY, Lu QM, Zhang XL (2007). Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells. World J Gastroenterol, 13, 1170-4.
  20. Michels J, Johnson PW, Packham G (2005). Mcl-1. Int J Biochem Cell Biol, 37, 267-71.
  21. Montecucco A, Biamonti G (2007). Cellular response to etoposide treatment. Cancer Lett, 252, 9-18.
  22. Nakao K, Hamasaki K, Ichikawa T, et al (2006). Survivin downregulation by siRNA sensitizes human hepatoma cells to TRAIL-induced apoptosis. Oncol Rep, 16, 389-92.
  23. Paduano F, Villa R, Pennati M, et al (2006). Silencing of survivin gene by small interfering RNAs produces supra-additive growth suppression in combination with 17-allylamino-17-demethoxygeldanamycin in human prostate cancer cells. Mol Cancer Ther, 5, 179-86.
  24. Robak T, Wierzbowska A (2009). Current and emerging therapies for acute myeloid leukemia. Clin Ther, 31, 2349-70.
  25. Ryan BM, O'Donovan N, Duffy MJ (2009). Survivin: a new target for anti-cancer therapy. Cancer Treat Rev, 35, 553-62.
  26. Shan G (2010). RNA interference as a gene knockdown technique. Int J Biochem Cell Biol, 42, 1243-51.
  27. Song H, Xin XY, Xiao F, at al (2008). Survivin gene RNA interference inhibits proliferation, induces apoptosis, and enhances radiosensitivity in HeLa cells. Eur J Obstet Gynecol Reprod Biol, 136, 83-9.
  28. Wacheck V, Cejka D, Sieghart W, et al (2006). Mcl-1 is a relevant molecular target for antisense oligonucleotide strategies in gastric cancer cells. Cancer Biol Ther, 5, 1348-54.
  29. Yang M, Mattes J (2008). Discovery, biology and therapeutic potential of RNA interference, microRNA and antagomirs. Pharmacol Ther, 117, 94-104.
  30. Zaffaroni N, Daidone MG (2002). Survivin expression and resistance to anticancer treatments: perspectives for new therapeutic interventions. Drug Resist Updat, 5, 65-72.
  31. Zaffaroni N, Pennati M, Daidone MG (2005). Survivin as a target for new anticancer interventions. J Cell Mol Med, 9, 360-72.

Cited by

  1. BCR/ABL mRNA Targeting Small Interfering RNA Effects on Proliferation and Apoptosis in Chronic Myeloid Leukemia vol.15, pp.12, 2014,
  2. Knowledge Production Status of Iranian Researchers in the Gastric Cancer Area: Based on the Medline Database vol.15, pp.12, 2014,
  3. MEKK3 and Survivin Expression in Cervical Cancer: Association with Clinicopathological Factors and Prognosis vol.15, pp.13, 2014,
  4. Elevated Serum Ferritin Levels in Patients with Hematologic Malignancies vol.15, pp.15, 2014,
  5. Lack of Relation of Survivin Gene Expression with Survival and Surgical Prognostic Factors in Endometrial Carcinoma Patients vol.15, pp.16, 2014,
  6. Upregulation of HIF-1α by Hypoxia Protect Neuroblastoma Cells from Apoptosis by Promoting Survivin Expression vol.15, pp.19, 2014,
  7. Telmisartan Exerts Anti-Tumor Effects by Activating Peroxisome Proliferator-Activated Receptor-γ in Human Lung Adenocarcinoma A549 Cells vol.19, pp.3, 2014,
  8. Knockdown of homeobox A5 by small hairpin RNA inhibits proliferation and enhances cytarabine chemosensitivity of acute myeloid leukemia cells vol.12, pp.5, 2015,
  9. Ki-67/MIB-1 as a Prognostic Marker in Cervical Cancer - a Systematic Review with Meta-Analysis vol.16, pp.16, 2015,
  10. Molecular Mechanisms of Apoptosis and Roles in Cancer Development and Treatment vol.16, pp.6, 2015,
  11. Piperlongumine reverses doxorubicin resistance through the PI3K/Akt signaling pathway in K562/A02 human leukemia cells vol.9, pp.4, 2015,
  12. Survivin: a unique target for tumor therapy vol.16, pp.1, 2016,
  13. Patient Derived Therapeutic Efficacy vol.14, pp.3, 2017,
  14. Inhibitors of apoptosis: clinical implications in cancer vol.22, pp.12, 2017,
  15. siRNA-Mediated Silencing of HMGA2 Induces Apoptosis and Cell Cycle Arrest in Human Colorectal Carcinoma vol.48, pp.2, 2017,
  16. Potential Molecular Targets in the Treatment of Lung Cancer Using siRNA Technology vol.36, pp.1, 2018,
  17. Glioblastoma and acute myeloid leukemia: malignancies with striking similarities vol.136, pp.2, 2018,