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Erschienen in: Cancer Chemotherapy and Pharmacology 3/2009

01.07.2009 | Short Communication

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

verfasst von: Jonathan T. Sims, Rina Plattner

Erschienen in: Cancer Chemotherapy and Pharmacology | Ausgabe 3/2009

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Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.
Literatur
1.
Zurück zum Zitat Pendergast AM (2002) The Abl family kinases: mechanisms of regulation and signaling. Adv Cancer Res 85:51–100PubMedCrossRef Pendergast AM (2002) The Abl family kinases: mechanisms of regulation and signaling. Adv Cancer Res 85:51–100PubMedCrossRef
2.
Zurück zum Zitat 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 19–39 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 19–39
3.
Zurück zum Zitat 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 344:1031–1037PubMedCrossRef 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 344:1031–1037PubMedCrossRef
4.
Zurück zum Zitat 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 2:561–566PubMedCrossRef 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 2:561–566PubMedCrossRef
5.
Zurück zum Zitat 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 13:2400–2411PubMedCrossRef 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 13:2400–2411PubMedCrossRef
6.
Zurück zum Zitat 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 5:309–319PubMedCrossRef 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 5:309–319PubMedCrossRef
7.
Zurück zum Zitat Srinivasan D, Plattner R (2006) Activation of abl tyrosine kinases promotes invasion of aggressive breast cancer cells. Cancer Res 66:5648–5655PubMedCrossRef Srinivasan D, Plattner R (2006) Activation of abl tyrosine kinases promotes invasion of aggressive breast cancer cells. Cancer Res 66:5648–5655PubMedCrossRef
8.
Zurück zum Zitat 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 27:1095–1105PubMedCrossRef 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 27:1095–1105PubMedCrossRef
9.
Zurück zum Zitat 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 69:581–593PubMedCrossRef 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 69:581–593PubMedCrossRef
10.
Zurück zum Zitat 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 303:474–482PubMedCrossRef 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 303:474–482PubMedCrossRef
11.
Zurück zum Zitat 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 195:193–199PubMed 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 195:193–199PubMed
12.
Zurück zum Zitat 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 23:181–188PubMedCrossRef 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 23:181–188PubMedCrossRef
13.
Zurück zum Zitat 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:1573–1577PubMedCrossRef 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:1573–1577PubMedCrossRef
14.
Zurück zum Zitat 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 16:481–486PubMed 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 16:481–486PubMed
15.
Zurück zum Zitat 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 104:13–19PubMedCrossRef 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 104:13–19PubMedCrossRef
16.
Zurück zum Zitat Burton EA, Plattner R, Pendergast AM (2003) Abl tyrosine kinases are required for infection by Shigella flexneri. EMBO J 22:5471–5479PubMedCrossRef Burton EA, Plattner R, Pendergast AM (2003) Abl tyrosine kinases are required for infection by Shigella flexneri. EMBO J 22:5471–5479PubMedCrossRef
17.
Zurück zum Zitat 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 16:127–135PubMedCrossRef 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 16:127–135PubMedCrossRef
18.
Zurück zum Zitat 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 50:167–169PubMedCrossRef 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 50:167–169PubMedCrossRef
19.
Zurück zum Zitat 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 5:69–84PubMed 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 5:69–84PubMed
20.
Zurück zum Zitat 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 44:1957–1962PubMedCrossRef 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 44:1957–1962PubMedCrossRef
Metadaten
Titel
MTT assays cannot be utilized to study the effects of STI571/Gleevec on the viability of solid tumor cell lines
verfasst von
Jonathan T. Sims
Rina Plattner
Publikationsdatum
01.07.2009
Verlag
Springer-Verlag
Erschienen in
Cancer Chemotherapy and Pharmacology / Ausgabe 3/2009
Print ISSN: 0344-5704
Elektronische ISSN: 1432-0843
DOI
https://doi.org/10.1007/s00280-009-1004-y

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