Advertisement

Tumor Biology

, Volume 37, Issue 2, pp 2365–2378 | Cite as

A molecular and biophysical comparison of macromolecular changes in imatinib-sensitive and imatinib-resistant K562 cells exposed to ponatinib

Original Article

Abstract

Chronic myeloid leukemia (CML) is a type of hematological malignancy that is characterized by the generation of Philadelphia chromosome encoding BCR/ABL oncoprotein. Tyrosine kinase inhibitors (TKIs), imatinib, nilotinib, and dasatinib, are used for the frontline therapy of CML. Development of resistance against these TKIs in the patients bearing T315I mutation is a major obstacle in CML therapy. Ponatinib, the third-generation TKI, is novel drug that is effective even in CML patients with T315I mutation. The exact mechanism of ponatinib in CML has been still unknown. In this study, we aimed to determine the potential mechanisms and structural metabolic changes activated by ponatinib treatment in imatinib-sensitive K562 human CML cell lines and 3 μM-imatinib-resistant K562/IMA3 CML cell lines generated at our lab. Apoptotic and antiproliferative effects of ponatinib on imatinib-sensitive and 3 μM-imatinib-resistant K562/IMA3 CML cells were determined by proliferation and apoptosis assays. Additionally, the effects of ponatinib on macromolecules and lipid profiles were also analyzed using Fourier transform infrared spectroscopy (FTIR). Our results revealed that ponatinib inhibited cell proliferation and induced apoptosis as determined by loss of mitochondrial membrane potential, increased caspase-3 enzyme activity, and transfer of phosphatidylserine to the plasma membrane in both K562 and K562/IMA-3 cells. Furthermore, cell cycle analyses revealed that ponatinib arrested K562 and K562/IMA-3 cells at G1 phase. Moreover, ponatinib treatment created a more ordered nucleic acid structure in the resistant cells. Although the lipid to protein ratio increased in imatinib-sensitive K562 cells with a little decrease in the K562/IMA-3 cells, ponatinib treatment indicated significant changes in the lipid composition such as a significant increase in the cellular cholesterol amounts much more in the K562/IMA-3 cells than the sensitive counterparts. Unsaturation in lipids was higher in the resistant cells; however, increases in lipids without phosphate and the number of acyl chains were much higher in the K562 cells. Taken together, all these results showed powerful antiproliferative and apoptotic effects of ponatinib in both imatinib-sensitive and imatinib-resistant CML cells in a dose-dependent manner, and hence, the use of ponatinib for the treatment of TKI-resistant CML patients may be an effective treatment approach in the clinic. More importantly, these results showed that FTIR spectroscopy can detect drug-induced physiological changes in cancer drug resistance.

Keywords

Ponatinib Chronic myeloid leukemia (CML) Multidrug resistance (MDR) Imatinib Fourier transform infrared spectroscopy (FTIR) 

Notes

Acknowledgments

This study was partly supported by TUBITAK project number 107S317 to Y.B. We thank İzmir Institute of Technology, Bioengineering and Biotechnology Application and Research Center for their assistance.

Conflicts of interest

None

References

  1. 1.
    Cea M, Cagnetta A, Nencioni A, Gobbi M, Patrone F. New insights into biology of chronic myeloid leukemia: implications in therapy. Curr Cancer Drug Targets. 2013;13:711–23.CrossRefPubMedGoogle Scholar
  2. 2.
    Pavlovsky C, Kantarjian H, Cortes JE. First-line therapy for chronic myeloid leukemia: past, present, and future. Am J Hematol. 2009;84:287–93.CrossRefPubMedGoogle Scholar
  3. 3.
    O’Brien S, Radich JP, Abboud CN, Akhtari M, Altman JK, Berman E, et al. Chronic myelogenous leukemia, version 1.2014. J Natl Compr Cancer Netw. 2013;11:1327–40.Google Scholar
  4. 4.
    Kimura S, Ando T, Kojima K. Ever-advancing chronic myeloid leukemia treatment. Int J Clin Oncol. 2014;19:3–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Weisberg E, Manley PW, Cowan-Jacob SW, Hochhaus A, Griffin JD. Second generation inhibitors of BCR-ABL for the treatment of imatinib-resistant chronic myeloid leukaemia. Nat Rev Cancer. 2007;7:345–56.CrossRefPubMedGoogle Scholar
  6. 6.
    La Rosee P, Corbin AS, Stoffregen EP, Deininger MW, Druker BJ. Activity of the Bcr-Abl kinase inhibitor PD180970 against clinically relevant Bcr-Abl isoforms that cause resistance to imatinib mesylate (Gleevec, STI571). Cancer Res. 2002;62:7149–53.PubMedGoogle Scholar
  7. 7.
    O’Hare T, Shakespeare WC, Zhu X, Eide CA, Rivera VM, Wang F, et al. AP24534, a pan-BCR-ABL inhibitor for chronic myeloid leukemia, potently inhibits the T315I mutant and overcomes mutation-based resistance. Cancer Cell. 2009;16:401–12.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Jain P, Kantarjian H, Cortes J. Chronic myeloid leukemia: overview of new agents and comparative analysis. Curr Treat Options in Oncol. 2013;14:127–43.CrossRefGoogle Scholar
  9. 9.
    Dogan A, Ergen K, Budak F, Severcan F. Evaluation of disseminated candidiasis on an experimental animal model: a Fourier transform infrared study. Appl Spectrosc. 2007;61:199–203.CrossRefPubMedGoogle Scholar
  10. 10.
    Cakmak G, Togan I, Severcan F. 17Beta-estradiol induced compositional, structural and functional changes in rainbow trout liver, revealed by FT-IR spectroscopy: a comparative study with nonylphenol. Aquat Toxicol. 2006;77:53–63.CrossRefPubMedGoogle Scholar
  11. 11.
    Kneipp J, Lasch P, Baldauf E, et al. Detection of pathological molecular alterations in scrapie-infected hamster brain by Fourier transform infrared (FT-IR) spectroscopy. Biochim Biophys Acta. 2000;1501:189–99.CrossRefPubMedGoogle Scholar
  12. 12.
    Gaigneaux A, Ruysschaert JM, Goormaghtigh E. Infrared spectroscopy as a tool for discrimination between sensitive and multiresistant K562 cells. Eur J Biochem. 2002;269:1968–73.CrossRefPubMedGoogle Scholar
  13. 13.
    Le Gal JM, Morjani H, Manfait M. Ultrastructural appraisal of the multidrug resistance in K562 and LR73 cell lines from Fourier transform infrared spectroscopy. Cancer Res. 1993;53:3681–6.PubMedGoogle Scholar
  14. 14.
    Piskin O, Ozcan MA, Ozsan GH, Ates H, Demirkan F, Alacacioglu I, et al. Synergistic effect of imatinib mesylate and fludarabine combination on Philadelphia chromosome-positive chronic myeloid leukemia cell lines. Turk J Haematol. 2007;24:23–7.PubMedGoogle Scholar
  15. 15.
    Gokbulut AA, Apohan E, Baran Y. Resveratrol and quercetin-induced apoptosis of human 232B4 chronic lymphocytic leukemia cells by activation of caspase-3 and cell cycle arrest. Hematology. 2013;18(3):144–50.CrossRefPubMedGoogle Scholar
  16. 16.
    Baran Y, Bielawski J, Gunduz U, Ogretmen B. Targeting glucosylceramide synthase sensitizes imatinib-resistant chronic myeloid leukemia cells via endogenous ceramide accumulation. J Cancer Res Clin Oncol. 2011;137(10):1535–44.CrossRefPubMedGoogle Scholar
  17. 17.
    Goktas S, Baran Y, Ural AU, Yazici S, Aydur E, Basal S, et al. Proteasome inhibitor bortezomib increases radiation sensitivity in androgen independent human prostate cancer cells. Urology. 2010;75(4):793–8.CrossRefPubMedGoogle Scholar
  18. 18.
    Petkovic M, Vocks A, Müller M, Schiller J, Arnhold J. Comparison of different procedures for the lipid extraction from HL-60 cells: a MALDI-TOF mass spectrometric study. Z Naturforsch. 2005;60c:143–51.Google Scholar
  19. 19.
    Ceylan C, Karacicek B. Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes/tripsin çözelti, jel ve agregat formlarının organik çözgen içeren ve pH-tektiklenmiş faz geçişlerinde yapısal ve fonksiyonel incelenmesi. Turk J Biochem. 2015;40:81–7.Google Scholar
  20. 20.
    Baran Y, Ceylan C, Camgoz A. The roles of macromolecules in imatinib resistance of chronic myeloid leukemia cells by Fourier transform infrared spectroscopy. Biomed Pharmacother. 2013;67:221–7.CrossRefPubMedGoogle Scholar
  21. 21.
    Melin AM, Perromat A, Deleris G. Pharmacologic application of Fourier transform IR spectroscopy: in vivo toxicity of carbon tetrachloride on rat liver. Biopolymers. 2000;57:160–8.CrossRefPubMedGoogle Scholar
  22. 22.
    Nara M, Okazaki M, Kagi H. Infrared study of human serum very-low-density and low-density lipoproteins. Implication of esterified lipid C O stretching bands for characterizing lipoproteins. Chem Phys Lipids. 2002;117:1–6.CrossRefPubMedGoogle Scholar
  23. 23.
    Voortman G, Gerrits J, Altavilla M, Henning M, Van BL, Hessels J. Quantitative determination of faecal fatty acids and triglycerides by Fourier transform infrared analysis with a sodium chloride transmission flow cell. Clin Chem Lab Med. 2002;40:795–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Manoharan R, Baraga JJ, Rava RP, Dasari RR, Fitzmaurice M, Feld MS. Biochemical analysis and mapping of atherosclerotic human artery using FT-IR microspectroscopy. Atherosclerosis. 1993;103:181–93.CrossRefPubMedGoogle Scholar
  25. 25.
    Jackson M, Ramjiawan B, Hewko M, Mantsch HH. Infrared microscopic functional group mapping and spectral clustering analysis of hypercholesterolemic rabbit liver. Cell Mol Biol. 1998;44:89–98.PubMedGoogle Scholar
  26. 26.
    Wang JJ, Chi CW, Lin SY, Chern YT. Conformational changes in gastric carcinoma cell membrane protein correlated to cell viability after treatment with adamantyl maleimide. Anticancer Res. 1997;17:3473–7.PubMedGoogle Scholar
  27. 27.
    Wong PT, Wong RK, Caputo TA, Godwin TA, Rigas B. Infrared spectroscopy of exfoliated human cervical cells: evidence of extensive structural changes during carcinogenesis. Proc Natl Acad Sci U S A. 1991;88:10988–92.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Chiriboga L, Xie P, Vigorita V, Zarou D, Zakim D, Diem M. Infrared spectroscopy of human tissue. II.A comparative study of spectra of biopsies of cervical squamous epithelium and of exfoliated cervical cells. Biospectroscopy. 1998;4:55–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Ci YX, Gao TY, Feng J, Guo JQ. Fourier transform infrared spectroscopic characterization of human breast tissue: implications for breast cancer diagnosis. Appl Spectrosc. 1999;53:312–5.CrossRefGoogle Scholar
  30. 30.
    Dovbeshko GI, Gridina NY, Kruglova EB, Pashchuk OP. FTIR spectroscopy studies of nucleic acid damage. Talanta. 2000;53:233–46.CrossRefPubMedGoogle Scholar
  31. 31.
    Pavlovsky C, Kantarjian H, Cortes JE. First-line therapy for chronic myeloid leukemia: past, present, and future. Am J Hematol. 2009;84:287–93.CrossRefPubMedGoogle Scholar
  32. 32.
    Frazer R, Irvine AE, McMullin MF. Chronic myeloid leukaemia in the 21st century. Ulster Med J. 2007;76(1):8–17.PubMedPubMedCentralGoogle Scholar
  33. 33.
    Cortes JE, Kantarjian H, Shah NP, Bixby D, Mauro MJ, Flinn I, et al. Ponatinib in refractory Philadelphia chromosome-positive leukemias. N Engl J Med. 2012;367:2075–88.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Cortes JE, Kim DW, Pinilla-Ibarz J, Le Coutre PD, Chuah C, Nicolini FE, Paquette R, Apperley JF, DiPersio JF, Khoury HJ, Rea D, Talpaz M, DeAngelo DJ, Abruzzese E, Baccarani M, Mueller MC, Gambacorti-Passerini C, Wong S, Lustgarten S, Turner CD, Rivera VM, Clackson T, Haluska F, Kantarjian HM, The PACE Study Group. Initial findings from the PACE Trial: a pivotal phase 2 study of ponatinib in patients with CML and Ph+ALL resistant or intolerant to dasatinib or nilotinib, or with the T315I mutation. ASH Meeting 2012; Abstract 163.Google Scholar
  35. 35.
    Cassuto O, Dufies M, Jacquel A, Robert G, Ginet C, Dubois A, et al. All tyrosine kinase inhibitor-resistant chronic myelogenous cells are highly sensitive to ponatinib. Oncotarget. 2012;3:1557–65.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Takahashi H, French SW, Wong PT. Alterations in hepatic lipids and proteins by chronic ethanol intake: a high-pressure Fourier transform infrared spectroscopic study on alcoholic liver disease in the rat. Alcohol Clin Exp Res. 1991;15:219–23.CrossRefPubMedGoogle Scholar
  37. 37.
    Kim HH, Kim T, Kim E, Park JK, Park SJ, Joo H. The mitochondrial Warburg effect: a cancer enigma. Interdisciplinary Bio Central. 2009;1:1–7.Google Scholar
  38. 38.
    Severcan F, Toyran N, Kaptan N, Turan B. Fourier transform infrared study of the effect of diabetes on rat liver and heart tissues in the CH region. Talanta. 2000;53:55–9.CrossRefPubMedGoogle Scholar
  39. 39.
    Schuldes H, Dolderer JH, Zimmer G, Knobloch J, Bickeboller R, Jonas D, et al. Reversal of multidrug resistance and increase in plasma membrane fluidity in CHO cells with R-verapamil and bile salts. Eur J Cancer. 2001;37:660–7.CrossRefPubMedGoogle Scholar
  40. 40.
    Tsvetkova NM, Horvath I, Torok Z, Wolkers WF, Balogi Z, Shigapova N, et al. Small heat-shock proteins regulate membrane lipid polymorphism. Proc Natl Acad Sci U S A. 2002;99:13504–9.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Colagar A, Chaichi M, Khadjvand T. Fourier transform infrared microspectroscopy as a diagnostic tool for distinguishing between normal and malignant human gastric tissue. J Biosci. 2011;36:669–77.CrossRefPubMedGoogle Scholar
  42. 42.
    Kochan K, Maslak E, Chlopicki S, Baranska M. FT-IR imaging for quantitative determination of liver fat content in non-alcoholic fatty liver. Analyst. 2015;140(15):4997–5002.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Department of Molecular Biology and Geneticsİzmir Institute of TechnologyUrlaTurkey
  2. 2.Department of Food Engineeringİzmir Institute of TechnologyUrlaTurkey
  3. 3.Department of Molecular Biology and Genetics, Faculty of Life and Natural SciencesAbdullah Gul UniversityKayseriTurkey

Personalised recommendations