Pharmaceutical Research

, Volume 34, Issue 1, pp 175–184 | Cite as

Therapeutic Effects of AICAR and DOX Conjugated Multifunctional Nanoparticles in Sensitization and Elimination of Cancer Cells via Survivin Targeting

  • Cenk DagliogluEmail author
  • Burcu Okutucu
Research Paper



Resistance to chemotherapy is one of the major problems facing current cancer research. Enhancing tumor cell response to anticancer agents increases chemotherapeutic effectiveness. We have recently addressed this issue and reported on producing multifunctional nanoparticles (Fe3O4@SiO2(FITC)-FA/AICAR/DOX) aiming to overcome chemoresistance with synergetic effect of AICAR and DOX. In the present study, we demonstrated that these nanoparticles not only show enhanced cellular uptake and cytotoxic effect but can also show enhanced pro-apoptotic and anti-proliferative effects in five different tumor-derived cell lines (A549, HCT-116, HeLa, Jurkat and MIA PaCa-2).


The nanoparticles were examined by using flow cytometric analyses of apoptosis and cell cycle. In addition, we performed caspase-3 activity assay, which supported our flow cytometric data. Furthermore, we demonstrated the applicability of this approach in a variety of cancer types confirming the potential widespread utility of this approach.


With the concept of co-delivery of AICAR and DOX in the nanoparticle formulation, the use of AICAR against survivin (BIRC5) sensitized cancer cells to DOX chemotherapy which resulted in effective cancer cell elimination. These result showed that combination therapy involving both a molecularly targeted therapy and chemotherapeutic agent has the ability to retain and enhance therapeutic efficacy.


Fe3O4@SiO2(FITC)-FA/AICAR/DOX nanoparticles is superior to monotherapy via the synergetic effect of AICAR and DOX and also the nanoparticle formulation could overcome issues of toxicity with targeted therapy while maintaining the potent anticancer effects of AICAR and DOX.

Graphical Abstract

Apoptosis analysis of A549 cells by flow cytometry-based PE-annexin-V / 7-ADD double staining treated with low-dose (10 μg/ml) concentration of (1) Fe3O4@SiO2(FITC)-FA (2) Fe3O4@SiO2(FITC)-FA/AICAR, (3) Fe3O4@SiO2(FITC)-FA/DOX or (4) Fe3O4@SiO2(FITC)-FA/AICAR/DOX nanoparticles. Viable cells labelled with PE-annexin-V(-)/7-ADD(-), early apoptotic cells labelled with PE-annexin-V(+)/7-ADD(-) and apoptotic cells labelled with PE-annexin-V(+)/ 7-ADD(+) in flow cytometric graphics.


AICAR chemoresistance DOX multifunctional nanoparticles survivin 









Dulbecco’s modified Eagle medium




Folic acid


Fluorescence-activated cell sorting


Fetal bovine serum


Fluorescein isothiocyanate


Heat shock protein 90


Inhibitors of apoptosis proteins


Phosphate buffered saline




Propidium iodide


Roswell Park Memorial Institute-1640 medium



The authors would like to thank Professor Anne Frary at the Izmir Institute of Technology for proofreading of the manuscript and Özgür Yılmazer from Biotechnology and Bioengineering Center of Izmir Institute of Technology for technical assistance during flow cytometry reading.


  1. 1.
    Lockshin RA, Zakeri Z. Cell death in health and disease. J Cell Mol Med. 2007;11:1214–24.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Letai AG. Diagnosing and exploiting cancer’s addiction to blocks in apoptosis. Nat Rev Cancer. 2008;8:121–32.CrossRefPubMedGoogle Scholar
  3. 3.
    Fulda S, Vucic D. Targeting IAP proteins for therapeutic intervention in cancer. Nat Rev Drug Disc. 2012;11:109–24.CrossRefGoogle Scholar
  4. 4.
    Li F. Survivin study: what is the next wave? J Cell Physiol. 1971;2003:8–29.Google Scholar
  5. 5.
    Duffy MJ, O’Donovan N, Brennan DJ, Gallagher WM, Ryan BM. Survivin: a promising tumor biomarker. Cancer Lett. 2007;2491:49–60.CrossRefGoogle Scholar
  6. 6.
    Altieri DC. Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer. 2008;81:61–70.CrossRefGoogle Scholar
  7. 7.
    Pennati M, Folini M, Zaffaroni N. Targeting survivin in cancer therapy. Expert Opin Ther Targets. 2008;124:463–76.CrossRefGoogle Scholar
  8. 8.
    Pennati M, Folini M, Zaffaroni N. Targeting survivin in cancer therapy: fulfilled promises and open questions. Carcinogenesis. 2007;286:1133–9.CrossRefGoogle Scholar
  9. 9.
    Chantalat L, Skoufias DA, Kleman JP, Jung B, Dideberg O, Margolis RL. Crystal structure of human survivin reveals a bow tie-shaped dimer with two unusual alpha-helical extensions. Mol Cell. 2000;61:183–9.CrossRefGoogle Scholar
  10. 10.
    Ryan BM, O’Donovan N, Duffy MJ. Survivin: a new target for anti-cancer therapy. Cancer Treat Rev. 2009;35:553–62.CrossRefPubMedGoogle Scholar
  11. 11.
    Whitesell L, Lindquist SL. Hsp90 and the chaperoning of cancer. Nat Rev Cancer. 2005;5:761–72.CrossRefPubMedGoogle Scholar
  12. 12.
    Fortugno P, Beltrami E, Plescia J, Fontana J, Pradhan D, Marchisio PC, et al. Regulation of survivin function by Hsp90. Proc Natl Acad Sci U S A. 2003;100:13791–6.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Plescia J, Salz W, Xia F, Pennati M, Zaffaroni N, Daidone MG, et al. Rational design of Shepherdin, a novel anticancer agent. Cancer Cell. 2005;7:457–67.CrossRefPubMedGoogle Scholar
  14. 14.
    Meli M, Pennati M, Curto M, Daidone MG, Plescia J, Toba S, et al. Small-molecule targeting of heat shock protein 90 chaperone function: rational identification of a new anticancer lead. J Med Chem. 2006;49:7721–30.CrossRefPubMedGoogle Scholar
  15. 15.
    Kizek R, Adam V, Hrabeta J, Eckschlager T, Smutny S, Burda JV, et al. Anthracyclines and ellipticines as DNA-damaging anticancer drugs: recent advances. Pharmacol Ther. 2012;133:26–39.CrossRefPubMedGoogle Scholar
  16. 16.
    Smith L, Watson MB, O’Kane SL, Drew PJ, Lind MJ, Cawkwell L. The analysis of doxorubicin resistance in human breast cancer cells using antibody microarrays. Mol Cancer Ther. 2006;5:2115–20.CrossRefPubMedGoogle Scholar
  17. 17.
    Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 2004;56:185–229.CrossRefPubMedGoogle Scholar
  18. 18.
    Daglioglu C, Okutucu B. Synthesis and characterization of AICAR and DOX conjugated multifunctional nanoparticles as a platform for synergistic inhibition of cancer cell growth. Bioconjug Chem. 2016;27:1098–111.CrossRefPubMedGoogle Scholar
  19. 19.
    Zaffaroni N, Pennati M, Colella G, Perego P, Supino R, Gatti L, et al. Expression of the anti-apoptotic gene survivin correlates with taxol resistance in human ovarian cancer. Cell Mol Life Sci. 2002;59:1406–12.CrossRefPubMedGoogle Scholar
  20. 20.
    Van Geelen CM, De Vries EG, De Jong S. Lessons from TRAIL-resistance mechanisms in colorectal cancer cells: paving the road to patient-tailored therapy. Drug Resist Updat. 2004;7:345–58.CrossRefPubMedGoogle Scholar
  21. 21.
    Asanuma K, Moriai R, Yajima T. Survivin as a radio-resistance factor in pancreatic cancer. Jpn J Cancer Res. 2000;91:1204–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Olie RA, Simoes-Wust AP, Baumann B, Leech SH, Fabbro D, Stahel RA, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy. Cancer Res. 2000;60:2805–9.PubMedGoogle Scholar
  23. 23.
    Li F, Altieri DC. Transcriptional analysis of human survivin gene expression. Biochem J. 1999;344(Pt 2):305–11.PubMedPubMedCentralGoogle Scholar
  24. 24.
    Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC, et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature. 1998;396:580–4.CrossRefPubMedGoogle Scholar
  25. 25.
    Tamm I, Wang Y, Sausville E, Scudiero DA, Vigna N, Oltersdorf T, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, Caspases, and Anticancer Drugs. Cancer Res. 1998;58:5315–20.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Faculty of Science, Department of Molecular Biology and GeneticsIzmir Institute of TechnologyUrla/IzmirTurkey
  2. 2.Faculty of Science, Biochemistry DepartmentEge UniversityBornova/IzmirTurkey

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