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Evaluation of a Tetrazolium-based Semiautomated Colorimetric Assay: Assessment of Chemosensitivity Testing¹

NCI-Navy Medical Oncology Branch [J.C., A.F.G., J.D.M.] and Radiobiology Section, Radiation Oncology Branch [J.C., W.G.D., J.B.M.], National Cancer Institute and Naval Hospital, Bethesda, Maryland 20814

Drug sensitivity assays were performed using a variation of a colorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] assay on V79, CHO-AuxB1, CH^RC5, NCI-H460, and NCI-H249 cell lines following optimization of experimental conditions for each cell line. Results from this assay were compared with data assimilated simultaneously by clonogenic assay and by dye exclusion assay. Good correlation was observed using the CHO-AuxB1 cell line and the pleiotropic drug-resistant mutant CH^RC5, with similar degrees of relative resistance observed with both the MTT and clonogenic assays. Good correlation was observed between the clonogenic and MTT assays for 1-h drug exposures, although the MTT assay was more sensitive to vinblastine. In general, the clonogenic assay was more sensitive when continuous drug exposures were utilized, although this was primarily related to the increased drug exposure time. While the use of the MTT assay in drug sensitivity testing of primary tumor samples is limited, since contaminating normal cells may also reduce the tetrazolium, the MTT assay can be semiautomated, and therefore it offers a valid, simple method of assessing chemosensitivity in established cell lines.

¹ The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or the Department of Defense.

² To whom requests for reprints should be addressed, at Radiation Oncology Branch, National Cancer Institute, Bldg. 10, Room B3-B69, Bethesda, MD 20892.

Received 4/25/86. Revised 9/18/86. Accepted 10/30/86.

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				N. Wajapeyee and K. Somasundaram Cell Cycle Arrest and Apoptosis Induction by Activator Protein 2{alpha} (AP-2{alpha}) and the Role of p53 and p21WAF1/CIP1 in AP-2{alpha}-mediated Growth Inhibition J. Biol. Chem., December 26, 2003; 278(52): 52093 - 52101. [Abstract] [Full Text] [PDF]

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				J. Litz, P. Carlson, G. S. Warshamana-Greene, S. Grant, and G. W. Krystal Flavopiridol Potently Induces Small Cell Lung Cancer Apoptosis during S Phase in a Manner That Involves Early Mitochondrial Dysfunction Clin. Cancer Res., October 1, 2003; 9(12): 4586 - 4594. [Abstract] [Full Text] [PDF]

				R. D. James, D. A. Jones, W. Aalbersberg, and C. M. Ireland Naamidine A Intensifies the Phosphotransferase Activity of Extracellular Signal-regulated Kinases Causing A-431 Cells to Arrest in G1 Mol. Cancer Ther., August 1, 2003; 2(8): 747 - 751. [Abstract] [Full Text] [PDF]

				Z. Duan, Y. Duan, D. E. Lamendola, R. Z. Yusuf, R. Naeem, R. T. Penson, and M. V. Seiden Overexpression of MAGE/GAGE Genes in Paclitaxel/Doxorubicin-resistant Human Cancer Cell Lines Clin. Cancer Res., July 1, 2003; 9(7): 2778 - 2785. [Abstract] [Full Text] [PDF]

				R. S. Sawhney, B. Sharma, L. E. Humphrey, and M. G. Brattain Integrin {alpha}2 and Extracellular Signal-regulated Kinase Are Functionally Linked in Highly Malignant Autocrine Transforming Growth Factor-{alpha}-driven Colon Cancer Cells J. Biol. Chem., May 23, 2003; 278(22): 19861 - 19869. [Abstract] [Full Text] [PDF]

				A. D. Colevas, J. M. Brown, S. Hahn, J. Mitchell, K. Camphausen, and C. N. Coleman Development of Investigational Radiation Modifiers J Natl Cancer Inst, May 7, 2003; 95(9): 646 - 651. [Full Text] [PDF]

				O. S. Frankfurt and A. Krishan Microplate Screening for Apoptosis with Antibody to Single-Stranded DNA Distinguishes Anticancer Drugs from Toxic Chemicals J Biomol Screen, April 1, 2003; 8(2): 185 - 190. [Abstract] [PDF]

				C. Roca, L. Primo, D. Valdembri, A. Cividalli, P. Declerck, P. Carmeliet, P. Gabriele, and F. Bussolino Hyperthermia Inhibits Angiogenesis by a Plasminogen Activator Inhibitor 1-dependent Mechanism Cancer Res., April 1, 2003; 63(7): 1500 - 1507. [Abstract] [Full Text] [PDF]

				M. Huang, Y. Wang, S. B. Cogut, B. S. Mitchell, and L. M. Graves Inhibition of Nucleoside Transport by Protein Kinase Inhibitors J. Pharmacol. Exp. Ther., February 1, 2003; 304(2): 753 - 760. [Abstract] [Full Text] [PDF]

				J.-W. Hyun, Y.-C. Jung, H.-S. Kim, E.-Y. Choi, J.-E. Kim, B.-H. Yoon, S.-H. Yoon, Y.-S. Lee, J. Choi, H.-J. You, et al. 8-Hydroxydeoxyguanosine Causes Death of Human Leukemia Cells Deficient in 8-Oxoguanine Glycosylase 1 Activity by Inducing Apoptosis Mol. Cancer Res., February 1, 2003; 1(4): 290 - 299. [Abstract] [Full Text] [PDF]

				M. Hayashi, M.-C. Rho, A. Enomoto, A. Fukami, Y.-P. Kim, Y. Kikuchi, T. Sunazuka, T. Hirose, K. Komiyama, and S. Omura Suppression of bone resorption by madindoline A, a novel nonpeptide antagonist to gp130 PNAS, November 12, 2002; 99(23): 14728 - 14733. [Abstract] [Full Text] [PDF]

				M. Hayashi, M.-C. Rho, A. Fukami, A. Enomoto, S. Nonaka, Y. Sekiguchi, T. Yanagisawa, A. Yamashita, T. Nogawa, Y. Kamano, et al. Biological Activity of a Novel Nonpeptide Antagonist to the Interleukin-6 Receptor 20S,21-Epoxy-resibufogenin-3-formate J. Pharmacol. Exp. Ther., October 1, 2002; 303(1): 104 - 109. [Abstract] [Full Text] [PDF]

				S. M. Hussain and J. M. Frazier Cellular Toxicity of Hydrazine in Primary Rat Hepatocytes Toxicol. Sci., October 1, 2002; 69(2): 424 - 432. [Abstract] [Full Text] [PDF]

				O. Volpert, W. Luo, T.-J. Liu, V. T. Estrera, C. Logothetis, and S.-H. Lin Inhibition of Prostate Tumor Angiogenesis by the Tumor Suppressor CEACAM1 J. Biol. Chem., September 13, 2002; 277(38): 35696 - 35702. [Abstract] [Full Text] [PDF]

				G. W. Krystal, G. Sulanke, and J. Litz Inhibition of Phosphatidylinositol 3-Kinase-Akt Signaling Blocks Growth, Promotes Apoptosis, and Enhances Sensitivity of Small Cell Lung Cancer Cells to Chemotherapy Mol. Cancer Ther., September 1, 2002; 1(11): 913 - 922. [Abstract] [Full Text] [PDF]

				D. Machover, J. Zittoun, R. Saffroy, P. Broet, S. Giraudier, T. Magnaldo, E. Goldschmidt, B. Debuire, M. Orrico, Y. Tan, et al. Treatment of Cancer Cells with Methioninase Produces DNA Hypomethylation and Increases DNA Synthesis Cancer Res., August 15, 2002; 62(16): 4685 - 4689. [Abstract] [Full Text] [PDF]

				A. Bandyopadhyay, F. Lopez-Casillas, S. N. Malik, J. L. Montiel, V. Mendoza, J. Yang, and L-Z. Sun Antitumor Activity of a Recombinant Soluble Betaglycan in Human Breast Cancer Xenograft Cancer Res., August 15, 2002; 62(16): 4690 - 4695. [Abstract] [Full Text] [PDF]

				S. Trohalaki, R. J. Zellmer, R. Pachter, S. M. Hussain, and J. M. Frazier Risk Assessment of High-Energy Chemicals by in Vitro Toxicity Screening and Quantitative Structure-Activity Relationships Toxicol. Sci., August 1, 2002; 68(2): 498 - 507. [Abstract] [Full Text] [PDF]

				T. S. Worgall, R. A. Johnson, T. Seo, H. Gierens, and R. J. Deckelbaum Unsaturated Fatty Acid-mediated Decreases in Sterol Regulatory Element-mediated Gene Transcription Are Linked to Cellular Sphingolipid Metabolism J. Biol. Chem., February 1, 2002; 277(6): 3878 - 3885. [Abstract] [Full Text] [PDF]

				R. S. Sawhney, G.-H. K. Zhou, L. E. Humphrey, P. Ghosh, J. I. Kreisberg, and M. G. Brattain Differences in Sensitivity of Biological Functions Mediated by Epidermal Growth Factor Receptor Activation with Respect to Endogenous and Exogenous Ligands J. Biol. Chem., January 4, 2002; 277(1): 75 - 86. [Abstract] [Full Text]

				X. Gao, A. T. Dinkova-Kostova, and P. Talalay Powerful and prolonged protection of human retinal pigment epithelial cells, keratinocytes, and mouse leukemia cells against oxidative damage: The indirect antioxidant effects of sulforaphane PNAS, December 18, 2001; 98(26): 15221 - 15226. [Abstract] [Full Text] [PDF]

				B. M. Sasseen, B. D. Gray, D. Gal, R. Lorinc, D. C. Carpenter, B. D. Klugherz, and R. L. Wilensky Local Delivery of a Hydrophobic Heparin Reduces Neointimal Hyperplasia After Balloon Injury in Rat Carotid but not Pig Coronary Arteries Journal of Cardiovascular Pharmacology and Therapeutics, December 1, 2001; 6(4): 377 - 383. [Abstract] [PDF]

				J. WITOWSKI, J. WISNIEWSKA, K. KORYBALSKA, T. O. BENDER, A. BREBOROWICZ, G. M. GAHL, U. FREI, J. PASSLICK-DEETJEN, and A. JORRES Prolonged Exposure to Glucose Degradation Products Impairs Viability and Function of Human Peritoneal Mesothelial Cells J. Am. Soc. Nephrol., November 1, 2001; 12(11): 2434 - 2441. [Abstract] [Full Text] [PDF]

				K. Tamagawa, K. Shimizu, T. Ebine, Y. Maitani, T. Fukui, K.-i. Kawai, and N. Takahashi Antitumor Efficacy in Vitro and in Vivo of Falconensones, a New Type of Polyene Clin. Cancer Res., November 1, 2001; 7(11): 3551 - 3558. [Abstract] [Full Text] [PDF]

				J. W. Fisher and J. Brookins Adenosine A2A and A2B receptor activation of erythropoietin production Am J Physiol Renal Physiol, November 1, 2001; 281(5): F826 - F832. [Abstract] [Full Text] [PDF]