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Feasibility of Drug Screening with Panels of Human Tumor Cell Lines Using a Microculture Tetrazolium Assay¹

Program Resources, Inc., National Cancer Institute-Frederick Cancer Research Facility, Frederick, Maryland 21701 [M. C. A., D. A. S., A. M., M. L. H., M. J. C., D. L. F.] and Developmental Therapeutics Program, Division of Cancer Treatment, National Cancer Institute, Bethesda, Maryland 20892 [B. J. A., J. G. M., R. H. S., M. R. B.]

For the past 30 years strategies for the preclinical discovery and development of potential anticancer agents have been based largely upon the testing of agents in mice bearing transplantable leukemias and solid tumors derived from a limited number of murine as well as human sources. The feasibility of implementing an alternate approach, namely combined in vitro/in vivo screening for selective cytotoxicity among panels of human tumor cell lines derived from a broad spectrum of human solid tumors is under investigation. A group of 30 cell lines acquired from a variety of sources and representing 8 lung cancer pathologies as well as 76 cell lines representing 10 other categories of human cancer (carcinomas of colon, breast, kidney, prostate, ovary, head and neck; glioma; leukemia; melanoma; and sarcoma) have exhibited acceptable growth characteristics and suitable colorimetric profiles in a single, standard culture medium. Measurements of in vitro growth in microculture wells by cell-mediated reduction of tetrazolium showed excellent correlation (0.89 < r² < 0.98) with measurements of cellular protein in adherent cell line cultures as well as viable cell count in suspension cell line cultures (0.94 < r² < 0.99). Since the microculture tetrazolium assay provides sensitive and reproducible indices of growth as well as drug sensitivity in individual cell lines over the course of multiple passages and several months' cultivation, it appears suitable for initial-stage in vitro drug screening.

¹ Supported by National Cancer Institute contract NO1-CO-23910 with Program Resources, Inc. The contents of this publication do not necessarily reflect the views or policies of the Department of Health and Human Services nor does mention of trade names, commercial products, or organizations imply endorsement by the U. S. Government.

² To whom requests for reprints should be addressed at: PDRG, Developmental Therapeutics Program, National Cancer Institute-Frederick Cancer Research Facility, Building 560, Room 32-60, Frederick, MD 21701.

Received 2/ 2/87. Revised 6/18/87. Revised 10/ 6/87. Accepted 11/ 2/87.

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				M. Naito, Y. Matsuba, S. Sato, H. Hirata, and T. Tsuruo MS-209, a Quinoline-type Reversal Agent, Potentiates Antitumor Efficacy of Docetaxel in Multidrug-resistant Solid Tumor Xenograft Models Clin. Cancer Res., February 1, 2002; 8(2): 582 - 588. [Abstract] [Full Text] [PDF]

				K. Nakamura, H. Sugumi, A. Yamaguchi, T. Uenaka, Y. Kotake, T. Okada, J. Kamata, J. Niijima, T. Nagasu, N. Koyanagi, et al. Antitumor Activity of ER-37328, a Novel Carbazole Topoisomerase II Inhibitor Mol. Cancer Ther., January 1, 2002; 1(3): 169 - 175. [Abstract] [Full Text] [PDF]

				M. Gao, S. Fan, I. D. Goldberg, J. Laterra, R. N. Kitsis, and E. M. Rosen Hepatocyte Growth Factor/Scatter Factor Blocks the Mitochondrial Pathway of Apoptosis Signaling in Breast Cancer Cells J. Biol. Chem., December 7, 2001; 276(50): 47257 - 47265. [Abstract] [Full Text] [PDF]

				D. Conze, L. Weiss, P. S. Regen, A. Bhushan, D. Weaver, P. Johnson, and M. Rincon Autocrine Production of Interleukin 6 Causes Multidrug Resistance in Breast Cancer Cells Cancer Res., December 1, 2001; 61(24): 8851 - 8858. [Abstract] [Full Text] [PDF]

				J. Ciccolini, P. Cuq, A. Evrard, S. Giacometti, A. Pelegrin, C. Aubert, J.-P. Cano, and A. Iliadis Combination of Thymidine Phosphorylase Gene Transfer and Deoxyinosine Treatment Greatly Enhances 5-Fluorouracil Antitumor Activity in Vitro and in Vivo Mol. Cancer Ther., December 1, 2001; 1(2): 133 - 139. [Abstract] [Full Text] [PDF]

				L. He, C.-P. Huang Yang, and S. B. Horwitz Mutations in {beta}-Tubulin Map to Domains Involved in Regulation of Microtubule Stability in Epothilone- resistant Cell Lines Mol. Cancer Ther., November 1, 2001; 1(1): 3 - 10. [Abstract] [Full Text] [PDF]

				R.-q. Yuan, S. Fan, M. Achary, D. M. Stewart, I. D. Goldberg, and E. M. Rosen Altered Gene Expression Pattern in Cultured Human Breast Cancer Cells Treated with Hepatocyte Growth Factor/Scatter Factor in the Setting of DNA Damage Cancer Res., November 1, 2001; 61(21): 8022 - 8031. [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]

				A. Li, M. L. Varney, and R. K. Singh Expression of Interleukin 8 and Its Receptors in Human Colon Carcinoma Cells with Different Metastatic Potentials Clin. Cancer Res., October 1, 2001; 7(10): 3298 - 3304. [Abstract] [Full Text] [PDF]

				S Wildi, J Kleeff, H Maruyama, C A Maurer, M W Buchler, and M Korc Overexpression of activin A in stage IV colorectal cancer Gut, September 1, 2001; 49(3): 409 - 417. [Abstract] [Full Text] [PDF]

				K. Miki, M. Xu, A. Gupta, Y. Ba, Y. Tan, W. Al-Refaie, M. Bouvet, M. Makuuchi, A. R. Moossa, and R. M. Hoffman Methioninase Cancer Gene Therapy with Selenomethionine as Suicide Prodrug Substrate Cancer Res., September 1, 2001; 61(18): 6805 - 6810. [Abstract] [Full Text] [PDF]

				S. Matsubara and M. Ozawa Expression of {alpha}-catenin in {alpha}-catenin-deficient cells increases resistance to sphingosine-induced apoptosis J. Cell Biol., August 6, 2001; 154(3): 573 - 584. [Abstract] [Full Text] [PDF]

				S. Fan, Y. X. Ma, M. Gao, R.-Q. Yuan, Q. Meng, I. D. Goldberg, and E. M. Rosen The Multisubstrate Adapter Gab1 Regulates Hepatocyte Growth Factor (Scatter Factor)-c-Met Signaling for Cell Survival and DNA Repair Mol. Cell. Biol., August 1, 2001; 21(15): 4968 - 4984. [Abstract] [Full Text] [PDF]

				J. Pardo, P. Perez-Galan, S. Gamen, I. Marzo, I. Monleon, A. A. Kaspar, S. A. Susin, G. Kroemer, A. M. Krensky, J. Naval, et al. A Role of the Mitochondrial Apoptosis-Inducing Factor in Granulysin-Induced Apoptosis J. Immunol., August 1, 2001; 167(3): 1222 - 1229. [Abstract] [Full Text] [PDF]

				A. A. Kaspar, S. Okada, J. Kumar, F. R. Poulain, K. A. Drouvalakis, A. Kelekar, D. A. Hanson, R. M. Kluck, Y. Hitoshi, D. E. Johnson, et al. A Distinct Pathway of Cell-Mediated Apoptosis Initiated by Granulysin J. Immunol., July 1, 2001; 167(1): 350 - 356. [Abstract] [Full Text] [PDF]

				M. Fan, M. E. Goodwin, M. J. Birrer, and T. C. Chambers The c-Jun NH2-terminal Protein Kinase/AP-1 Pathway Is Required for Efficient Apoptosis Induced by Vinblastine Cancer Res., June 1, 2001; 61(11): 4450 - 4458. [Abstract] [Full Text] [PDF]

				S. C. Modesitt, P. Ramirez, Z. Zu, D. Bodurka-Bevers, D. Gershenson, and J. K. Wolf In Vitro and in Vivo Adenovirus-mediated p53 and p16 Tumor Suppressor Therapy in Ovarian Cancer : See The Biology Behind: M. E. Murphy, The Battle between Tumor Suppressors: Is Gene Therapy Using p16INK4a More Efficacious Than p53 for Treatment of Ovarian Carcinoma? Clin. Cancer Res., 7: 1487-1489, 2001. Clin. Cancer Res., June 1, 2001; 7(6): 1765 - 1772. [Abstract] [Full Text] [PDF]

				K. E. Zhang, E. Wu, A. K. Patick, B. Kerr, M. Zorbas, A. Lankford, T. Kobayashi, Y. Maeda, B. Shetty, and S. Webber Circulating Metabolites of the Human Immunodeficiency Virus Protease Inhibitor Nelfinavir in Humans: Structural Identification, Levels in Plasma, and Antiviral Activities Antimicrob. Agents Chemother., April 1, 2001; 45(4): 1086 - 1093. [Abstract] [Full Text]

				M. R. Johnston, J. B.M. Mullen, M. E. Pagura, and R. B. Howard Validation of an orthotopic model of human lung cancer with regional and systemic metastases Ann. Thorac. Surg., April 1, 2001; 71(4): 1120 - 1125. [Abstract] [Full Text] [PDF]

				M. Fan, L. Du, A. A. Stone, K. M. Gilbert, and T. C. Chambers Modulation of Mitogen-activated Protein Kinases and Phosphorylation of Bcl-2 by Vinblastine Represent Persistent Forms of Normal Fluctuations at G2-M Cancer Res., November 1, 2000; 60(22): 6403 - 6407. [Abstract] [Full Text]

				H. J. Lim, M. J. Parr, D. Masin, N. L. McIntosh, T. D. Madden, G. Zhang, S. Johnstone, and M. B. Bally Kupffer Cells Do Not Play a Role in Governing the Efficacy of Liposomal Mitoxantrone Used to Treat a Tumor Model Designed to Assess Drug Delivery to Liver Clin. Cancer Res., November 1, 2000; 6(11): 4449 - 4460. [Abstract] [Full Text] [PDF]

				S. Fan, Q. Meng, B. Gao, J. Grossman, M. Yadegari, I. D. Goldberg, and E. M. Rosen Alcohol Stimulates Estrogen Receptor Signaling in Human Breast Cancer Cell Lines Cancer Res., October 1, 2000; 60(20): 5635 - 5639. [Abstract] [Full Text]

				S. Huang, A. DeGuzman, C. D. Bucana, and I. J. Fidler Nuclear Factor-{{kappa}}B Activity Correlates with Growth, Angiogenesis, and Metastasis of Human Melanoma Cells in Nude Mice Clin. Cancer Res., June 1, 2000; 6(6): 2573 - 2581. [Abstract] [Full Text]

				K. Miki, W. Al-Refaie, M. Xu, P. Jiang, Y. Tan, M. Bouvet, M. Zhao, A. Gupta, T. Chishima, H. Shimada, et al. Methioninase Gene Therapy of Human Cancer Cells Is Synergistic with Recombinant Methioninase Treatment Cancer Res., May 1, 2000; 60(10): 2696 - 2702. [Abstract] [Full Text]

				D. Vandier, V. Calvez, L. Massade, A. Gouyette, L. Mickley, T. Fojo, and O. Rixe Transactivation of the Metallothionein Promoter in Cisplatin-Resistant Cancer Cells: a Specific Gene Therapy Strategy J Natl Cancer Inst, April 19, 2000; 92(8): 642 - 647. [Abstract] [Full Text] [PDF]

				J. Ciccolini, L. Peillard, A. Evrard, P. Cuq, C. Aubert, A. Pelegrin, P. Formento, G. Milano, and J. Catalin Enhanced Antitumor Activity of 5-Fluorouracil in Combination with 2'-Deoxyinosine in Human Colorectal Cell Lines and Human Colon Tumor Xenografts Clin. Cancer Res., April 1, 2000; 6(4): 1529 - 1535. [Abstract] [Full Text]

				S. Poujol, F. Pinguet, F. Bressole, M. Boustta, and M. Vert Molecular Microencapsulation: Paclitaxel Formations in Aqueous Medium Using Hydrophobized Poly(L-Lysine Citramide Imide) Journal of Bioactive and Compatible Polymers, March 1, 2000; 15(2): 99 - 114. [Abstract] [PDF]

				L. Ramondetta, G. B. Mills, T. W. Burke, and J. K. Wolf Adenovirus-mediated Expression of p53 or p21 in a Papillary Serous Endometrial Carcinoma Cell Line (SPEC-2) Results in Both Growth Inhibition and Apoptotic Cell Death: Potential Application of Gene Therapy to Endometrial Cancer Clin. Cancer Res., January 1, 2000; 6(1): 278 - 284. [Abstract] [Full Text]

				B. Tang, K. de Castro, H. E. Barnes, W. T. Parks, L. Stewart, E. P. Bottinger, D. Danielpour, and L. M. Wakefield Loss of Responsiveness to Transforming Growth Factor {beta} Induces Malignant Transformation of Nontumorigenic Rat Prostate Epithelial Cells Cancer Res., October 1, 1999; 59(19): 4834 - 4842. [Abstract] [Full Text] [PDF]