Cancer Chemotherapy and Pharmacology

, Volume 64, Issue 3, pp 629–633 | Cite as

MTT assays cannot be utilized to study the effects of STI571/Gleevec on the viability of solid tumor cell lines

  • Jonathan T. Sims
  • Rina PlattnerEmail author
Short Communication



This study will determine whether MTT assays accurately assess the effect of STI571 (Gleevec; Abl kinase inhibitor) on the viability of cancer cells containing highly active Abl kinases.


Growth kinetics, tritiated thymidine, fluorescent caspase, MTT, and Cell Titer Glo (CTG) assays were used to determine the effect of STI571 on growth, proliferation, apoptosis, and viability of melanoma and breast cancer cells.


STI571 inhibited growth and proliferation, and increased apoptosis. However, MTT assays indicated that STI571 increased cell viability. In contrast, STI571 induced a dose-dependent decrease in viability using CTG assays.


Doses of STI571 (1–10 μM) required to inhibit endogenous Abl kinases interfere with the MTT assay, and therefore MTT cannot be used to determine the effect of STI571 on viability using these doses. Additionally, caution should be utilized when interpreting the results of MTT assays used to screen kinase inhibitors for anti-cancer activity, as drug effectiveness may be minimized.


STI571 Gleevec Abl MTT Cell Titer Glo (CTG) Viability 



This work was supported by National Institute of Health Grant P20 RR20171 from the National Center for Research Resources, and National Institute of Health/National Cancer Institute Grant 1R01CA116784 to R.P. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIH. We thank Elisabeth Buchdunger (Novartis Pharmaceuticals, Basel, Switzerland) for providing STI571, and Leann Fiore for critically reading the manuscript.


  1. 1.
    Pendergast AM (2002) The Abl family kinases: mechanisms of regulation and signaling. Adv Cancer Res 85:51–100PubMedCrossRefGoogle Scholar
  2. 2.
    Pendergast AM (2001) BCR-ABL protein domain, function, and signaling. In: Helmann R (ed) Chronic myeloid leukaemia: biology and treatment. Martin Dunitz Lt, London, pp 19–39Google Scholar
  3. 3.
    Druker BJ, Talpaz M, Resta DJ, Peng B, Buchdunger E, Ford JM, Lydon NB, Kantarjian H, Capdeville R, Ohno-Jones S, Sawyers CL (2001) Efficacy and safety of a specific inhibitor of the BCR-ABL tyrosine kinase in chronic myeloid leukemia. N Engl J Med 344:1031–1037PubMedCrossRefGoogle Scholar
  4. 4.
    Druker BJ, Tamura S, Buchdunger E, Ohno S, Segal GM, Fanning S, Zimmermann J, Lydon NB (1996) Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med 2:561–566PubMedCrossRefGoogle Scholar
  5. 5.
    Plattner R, Kadlec L, DeMali KA, Kazlauskas A, Pendergast AM (1999) c-Abl is activated by growth factors and Src family kinases and has a role in the cellular response to PDGF. Genes Dev 13:2400–2411PubMedCrossRefGoogle Scholar
  6. 6.
    Plattner R, Irvin BJ, Guo S, Blackburn K, Kazlauskas A, Abraham RT, York JD, Pendergast AM (2003) A new link between the c-Abl tyrosine kinase and phosphoinositide signaling via PLC-γ1. Nat Cell Biol 5:309–319PubMedCrossRefGoogle Scholar
  7. 7.
    Srinivasan D, Plattner R (2006) Activation of abl tyrosine kinases promotes invasion of aggressive breast cancer cells. Cancer Res 66:5648–5655PubMedCrossRefGoogle Scholar
  8. 8.
    Srinivasan D, Sims JT, Plattner R (2008) Aggressive breast cancer cells are dependent on activated Abl kinases for proliferation, anchorage-independent growth and survival. Oncogene 27:1095–1105PubMedCrossRefGoogle Scholar
  9. 9.
    Liu Y, Peterson DA, Kimura H, Schubert D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem 69:581–593PubMedCrossRefGoogle Scholar
  10. 10.
    Berridge MV, Tan AS (1993) Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch Biochem Biophys 303:474–482PubMedCrossRefGoogle Scholar
  11. 11.
    Bernhard D, Schwaiger W, Crazzolara R, Tinhofer I, Kofler R, Csordas A (2003) Enhanced MTT-reducing activity under growth inhibition by resveratrol in CEM-C7H2 lymphocytic leukemia cells. Cancer Lett 195:193–199PubMedGoogle Scholar
  12. 12.
    Vellonen KS, Honkakoski P, Urtti A (2004) Substrates and inhibitors of efflux proteins interfere with the MTT assay in cells and may lead to underestimation of drug toxicity. Eur J Pharm Sci 23:181–188PubMedCrossRefGoogle Scholar
  13. 13.
    Pagliacci MC, Spinozzi F, Migliorati G, Fumi G, Smacchia M, Grignani F, Riccardi C, Nicoletti I (1993) Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: a further pitfall in the use of the MTT assay for evaluating cell growth and survival. Eur J Cancer 29A:1573–1577PubMedCrossRefGoogle Scholar
  14. 14.
    Es-Saady D, Simon A, Jayat-Vignoles C, Chulia AJ, Delage C (1996) MCF-7 cell cycle arrested at G1 through ursolic acid, and increased reduction of tetrazolium salts. Anticancer Res 16:481–486PubMedGoogle Scholar
  15. 15.
    Rae JM, Creighton CJ, Meck JM, Haddad BR, Johnson MD (2007) MDA-MB-435 cells are derived from M14 melanoma cells—a loss for breast cancer, but a boon for melanoma research. Breast Cancer Res Treat 104:13–19PubMedCrossRefGoogle Scholar
  16. 16.
    Burton EA, Plattner R, Pendergast AM (2003) Abl tyrosine kinases are required for infection by Shigella flexneri. EMBO J 22:5471–5479PubMedCrossRefGoogle Scholar
  17. 17.
    Mayorga ME, Sanchis D, Perez de Santos AM, Velasco A, Dolcet X, Casanova JM, Baradad M, Egido R, Pallares J, Espurz N, Benitez D, Mila J, Malvehy J, Castel T, Comella JX, Matias-Guiu X, Vilella R, Marti RM (2006) Antiproliferative effect of STI571 on cultured human cutaneous melanoma-derived cell lines. Melanoma Res 16:127–135PubMedCrossRefGoogle Scholar
  18. 18.
    Hotfilder M, Lanvers C, Jurgens H, Boos J, Vormoor J (2002) c-KIT-expressing Ewing tumour cells are insensitive to imatinib mesylate (STI571). Cancer Chemother Pharmacol 50:167–169PubMedCrossRefGoogle Scholar
  19. 19.
    Marshall NJ, Goodwin CJ, Holt SJ (1995) A critical assessment of the use of microculture tetrazolium assays to measure cell growth and function. Growth Regul 5:69–84PubMedGoogle Scholar
  20. 20.
    Hayon T, Dvilansky A, Shpilberg O, Nathan I (2003) Appraisal of the MTT-based assay as a useful tool for predicting drug chemosensitivity in leukemia. Leuk Lymphoma 44:1957–1962PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Molecular and Biomedical PharmacologyUniversity of Kentucky School of MedicineLexingtonUSA

Personalised recommendations