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Components of the Cell Death Machine and Drug Sensitivity of the National Cancer Institute Cell Line Panel

¹ Division of Oncology Research, Mayo Clinic, and ² Department of Molecular Pharmacology, Mayo Graduate School, Rochester, Minnesota; ³ Cold Spring Harbor Laboratory, Cold Spring Harbor, New York; ⁴ SAIC-Frederick, Inc., National Cancer Institute, NIH, Frederick, Maryland; ⁵ Developmental Therapeutics Program, National Cancer Institute, NIH, Bethesda, Maryland; and ⁶ Burnham Institute, LaJolla, California

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-hour 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 x 10⁵ to 2 x 10⁶ molecules per cell and procaspase-9 from 5 x 10³ to 1.6 x 10⁵ molecules per cell. Procaspase-8 levels ranged from 1.7 x 10⁵ to 8 x 10⁶ molecules per cell. Procaspase-3, a major effector caspase, varied from undetectable to 1.6 x 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.

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