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[Cancer Research 57, 4285-4300, October 1, 1997]
© 1997 American Association for Cancer Research
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Characterization of the p53 Tumor Suppressor Pathway in Cell Lines of the National Cancer Institute Anticancer Drug Screen and Correlations with the Growth-Inhibitory Potency of 123 Anticancer Agents

Patrick M. O'Connor1, Joany Jackman, Insoo Bae, Timothy G. Myers, Saijun Fan, Masato Mutoh, Dominic A. Scudiero, Ann Monks, Edward A. Sausville, John N. Weinstein, Stephen Friend, Albert J. Fornace, Jr. and Kurt W. Kohn

Laboratory of Molecular Pharmacology, Division of Basic Sciences, National Cancer Institute, NIH, Bethesda, Maryland 20892 [P. M. O., J. J., I. B., T. G. M., S. Fa., M. M., J. N. W., A. J. F., K. W. K.]; Developmental Therapeutics Program, National Cancer Institute, Frederick Cancer Research & Development Center, Frederick, Maryland 21702 [D. A. S., A. M., E. A. S.]; and Laboratory of Molecular Pharmacology, Fred Hutchinson Cancer Center, Seattle, Washington 98104 [S. Fr.]

In the present study, we report the characterization of the p53 tumor suppressor pathway in the 60 cell lines of the National Cancer Institute (NCI) anticancer drug screen, as well as correlations between the integrity of this pathway and the growth-inhibitory potency of 123 anticancer agents in this screen. Assessment of p53 status in these lines was achieved through complete p53 cDNA sequencing, measurement of basal p53 protein levels and functional assessment of (a) transcriptional activity of p53 cDNA from each line in a yeast assay, (b) {gamma}-ray-induced G1 phase cell cycle arrest, and (c) {gamma}-ray-induced expression of CIP1/WAF1, GADD45, and MDM2 mRNA. Our investigations revealed that p53 gene mutations were common in the NCI cell screen lines: 39 of 58 cell lines analyzed contained a mutant p53 sequence. cDNA derived from almost all of the mutant p53 cell lines failed to transcriptionally activate a reporter gene in yeast, and the majority of mutant p53 lines studied expressed elevated basal levels of the mutant p53 protein. In contrast to most of the wild-type p53-containing lines, cells containing mutant p53 sequence were also deficient in {gamma}-ray induction of CIP1/WAF1, GADD45, and MDM2 mRNA and the ability to arrest in G1 following {gamma}-irradiation. Taken together, these assessments provided indications of the integrity of the p53 pathway in the 60 cell lines of the NCI cell screen. These individual p53 assessments were subsequently used to probe a database of growth-inhibitory potency for 123 "standard agents," which included the majority of clinically approved anticancer drugs. These 123 agents have been tested against these lines on multiple occasions, and a proposed mechanism of drug action had previously been assigned to each agent. Our analysis revealed that cells with mutant p53 sequence tended to exhibit less growth inhibition in this screen than the wild-type p53 cell lines when treated with the majority of clinically used anticancer agents: including DNA cross-linking agents, antimetabolites, and topoisomerase I and II inhibitors. Similar correlations were uncovered when we probed this database using most of the other indices of p53 status we assessed in the lines. Interestingly, a class of agents that differed in this respect was the antimitotic agents. Growth-inhibitory activity of these agents tended, in this assay, to be independent of p53 status. Our characterization of the p53 pathway in the NCI cell screen lines should prove useful to researchers investigating fundamental aspects of p53 biology and pharmacology. This information also allows for the large-scale analysis of the more than 60,000 compounds tested against these lines for novel agents that might exploit defective p53 function as a means of preferential toxicity.

1 To whom requests for reprints should be addressed, at Laboratory of Molecular Pharmacology, Division of Basic Science, Building 37, Room 5C25, National Cancer Institute, NIH, Bethesda, MD 20892. Phone: (301) 435-2848; Fax: (301) 402-0752; E-mail: oconnorp@dc37a.nci.nih.gov.

Received 4/25/97. Accepted 7/25/97.




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X. Wu, W. Fan, S. Xu, and Y. Zhou
Sensitization to the Cytotoxicity of Cisplatin by Transfection with Nucleotide Excision Repair Gene Xeroderma Pigmentosun Group A Antisense RNA in Human Lung Adenocarcinoma Cells
Clin. Cancer Res., December 1, 2003; 9(16): 5874 - 5879.
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