Evaluation of Action Mechanisms of Toxic Chemicals Using JFCR39, a Panel of Human Cancer Cell Lines

Abstract

We previously established a panel of human cancer cell lines, JFCR39, coupled to an anticancer drug activity database; this panel is comparable with the NCI60 panel developed by the National Cancer Institute. The JFCR39 system can be used to predict the molecular targets or evaluate the action mechanisms of the test compounds by comparing their cell growth inhibition profiles (i.e., fingerprints) with those of the standard anticancer drugs using the COMPARE program. In this study, we used this drug activity database-coupled JFCR39 system to evaluate the action mechanisms of various chemical compounds, including toxic chemicals, agricultural chemicals, drugs, and synthetic intermediates. Fingerprints of 130 chemicals were determined and stored in the database. Sixty-nine of 130 chemicals (ϳ60%) satisfied our criteria for the further analysis and were classified by cluster analysis of the fingerprints of these chemicals and several standard anticancer drugs into the following three clusters: 1) anticancer drugs, 2) chemicals that shared similar action mechanisms (for example, ouabain and digoxin), and 3) chemicals whose action mechanisms were unknown. These results suggested that chemicals belonging to a cluster (i.e., a cluster of toxic chemicals, a cluster of anticancer drugs, etc.) shared similar action mechanism. In summary, the JFCR39 system can classify chemicals based on their fingerprints, even when their action mechanisms are unknown, and it is highly probable that the chemicals within a cluster share common action mechanisms.Determining the action mechanism or identifying the molecular target of a chemical with pharmacological activity or adverse side effects is highly desirable. Although various test methods are currently available for determining the action mechanisms of chemicals, such as methods based on animal models, methods based on cellular models, bacterial mutagenicity test, the uterotropic assay (Kanno et al., 2002), Hershberger test (Hershberger et al., 1953), and the reporter assay for the nuclear receptor agonists, determination of the action mechanisms of pharmacologically active chemicals, including the toxic chemicals, is still a difficult and challenging task. Therefore, it is highly desirable to develop efficient test methods for evaluating toxicity of chemicals.A number of screening methods are currently available for discovering new anticancer drugs. One very powerful and unique approach using multiple cancer cell lines was developed at NCI (Paull et al

ZSTK474, a novel phosphatidylinositol 3-kinase inhibitor identified using the JFCR39 drug discovery system

“…system, an informatic anticancer drug discovery system for molecular target identification JFCR39 is an informatic anticancer drug screening system we established in the early 1990s by utilizing a human cancer cell line panel JFCR39 coupled with a drug-activity database [1][2][3][4] . This system was developed based on the NCI60 system [5] with some modifications [4] .…”

Effect of ZSTK474, a Novel Phosphatidylinositol 3-Kinase Inhibitor, on DNA-Dependent Protein Kinase

“…18) ZSTK474 is a novel PI3K inhibitor, identified in our laboratory by COMPARE analysis using a panel of 39 human cancer cell lines coupled to a drug-activity database. [19][20][21][22][23] ZSTK474 exhibited highly promising antitumor efficacy without observable toxicity. We previously reported that ZSTK474 inhibited all four PI3K isoforms, indicative of a pan-PI3K inhibitor, while retaining high selectivity for PI3K over 140 protein kinases.…”

Synthesis and Structure–Activity Relationship Study of NBRI16716B, an Antitumor Natural Product

“…The white powder was corrected on funnel, which was thoroughly washed with CH 3 CN and MeOH to give pure sample of 22 (14.9 Cells and Reagents Human lung cancer cell line A549 was obtained as described. 22) NHLF normal human lung fibroblasts were obtained from BioWhittaker. The stromal cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin G, 100 µg/mL streptomycin, ITH (5 µg/ mL insulin, 5 µg/mL transferrin, and 1.4 µM hydrocortisone), and 5 ng/mL basic-FGF (Pepro Tech, NJ, U.S.A.) at 37°C with 5% CO 2 as described. )…”