Components of the cell death machine and drug sensitivity of the National Cancer Institute cell line panel

Phyllis A. Svingen, David Loegering, Joe Rodriquez, Xue Wei Meng, Peter W. Mesner, Susan Holbeck, Anne Monks, Stan Krajewski, Dominic A. Scudiero, Edward A. Sausville, John C. Reed, Yuri A. Lazebnik, Scott H. Kaufmann

Research output: Contribution to journalArticle

56 Scopus citations

Abstract

Purpose: According to some studies, susceptibility of cells to anticancer drug-induced apoptosis is markedly inhibited by targeted deletion of genes encoding apoptotic protease activating factor 1 (Apaf-1) or certain caspases. Information about levels of these polypeptides in common cancer cell types and any possible correlation with drug sensitivity in the absence of gene deletion is currently fragmentary. Experimental Design: Immunoblotting was used to estimate levels of Apaf-1 as well as procaspase-2, -3, -6, -7, -8, and -9 in the 60-cell-line panel used for drug screening by the National Cancer Institute. Sensitivity of the same lines to >80,000 compounds was determined with 48-hoer sulforhodamine B binding assays. Additional 6-day assays were performed for selected agents. Results: Levels of Apaf-1 and procaspases varied widely. Apaf-1 and procaspase-9, which are implicated in caspase activation after treatment of cells with various anticancer drugs, were detectable in all of the cell lines, with levels of Apaf-1 ranging from ∼1 × 105 to 2 × 106 molecules per cell and procaspase-9 from ∼5 × 103 to ∼1.6 × 105 molecules per cell. Procaspase-8 levels ranged from 1.7 × 105 to 8 × 10 6 molecules per cell. Procaspase-3, a major effector caspase, varied from undetectable to ∼1.6 × 106 molecules per cell. Correlations between levels of these polypeptides and sensitivity to any of a variety of experimental or conventional antineoplastic agents in either 2-day or 6-day cytotoxicity assays were weak at best. Conclusions: With the exception of caspase-3, all of the components of the core cell-death machinery are expressed in all of the cell lines examined. Despite variations in expression, levels of any one component are not a major determinant of drug sensitivity in these cells in vitro.

Original languageEnglish (US)
Pages (from-to)6807-6820
Number of pages14
JournalClinical Cancer Research
Volume10
Issue number20
DOIs
StatePublished - Oct 15 2004

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Svingen, P. A., Loegering, D., Rodriquez, J., Meng, X. W., Mesner, P. W., Holbeck, S., Monks, A., Krajewski, S., Scudiero, D. A., Sausville, E. A., Reed, J. C., Lazebnik, Y. A., & Kaufmann, S. H. (2004). Components of the cell death machine and drug sensitivity of the National Cancer Institute cell line panel. Clinical Cancer Research, 10(20), 6807-6820. https://doi.org/10.1158/1078-0432.CCR-0778-02