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Preselection of Potential Cancerostatics by Automatic Analysis of Suspended and Adherent Cells Incubated in
1Hans Knöll Institute of Natural Product Research, Jena, Germany.
Abstract:
Alternative toxicological screening programmes, without the use of animal experiments, are intended to eliminate dangerous substances and to find new pharmacologically active agents in cell cultures. They can also provide information on the cytostatic activities of the agents. Intercalating cytostatics which bind DNA were selected by measuring the statistical distributions of the cell diameters of K-562 and L-929 cells by using an electronic cell analyser (CASY1). These compounds were identified by cell enlargement or from flat concentration-activity curves created with the cell analyser system. Incubation for 72 hours with DNA-binding agents, such as doxorubicin, daunorubicin and Mitoxantron®, resulted in enlargement of cell diameter and cell volume. The antineoplastic agents actinomycin D and ambazone had no comparable effect. Comparisons of the different parameters obtained with CASY1 measurement were performed with Microsoft EXCEL.
Insights
This study introduces an animal-free toxicological screening method using an electronic cell analyzer to detect DNA-binding cytostatics. The method identifies compounds causing cell enlargement, aiding in the discovery of novel pharmaceutical agents.
Area of Science:
- Toxicology
- Pharmacology
- Cell Biology
Background:
- Alternative toxicological screening programs aim to replace animal testing for identifying dangerous substances and discovering new drugs.
- Cell cultures offer a platform for evaluating pharmacological activity and cytostatic effects of agents.
- DNA-binding cytostatics are a class of antineoplastic agents with potential therapeutic applications.
Purpose of the Study:
- To develop and validate an alternative toxicological screening method without animal experiments.
- To identify DNA-binding cytostatic agents using an electronic cell analyzer.
- To assess the cytostatic activity and mechanism of action of selected compounds.
Main Methods:
- Utilized an electronic cell analyzer (CASY1) to measure cell diameter distributions in K-562 and L-929 cell lines.
- Selected intercalating, DNA-binding cytostatics based on cell enlargement or flat concentration-activity curves.
- Incubated cells for 72 hours with various DNA-binding agents and antineoplastic drugs.
- Analyzed and compared measurement parameters using Microsoft EXCEL.
Main Results:
- Doxorubicin, daunorubicin, and Mitoxantron® (DNA-binding agents) caused significant cell enlargement and increased cell volume.
- Actinomycin D and ambazone, other antineoplastic agents, did not produce comparable cell enlargement.
- The CASY1 system effectively differentiated compounds based on their effects on cell size and volume.
Conclusions:
- The electronic cell analyzer (CASY1) is a viable tool for alternative toxicological screening of DNA-binding cytostatics.
- Cell enlargement serves as a reliable indicator for identifying intercalating, DNA-binding agents.
- This method facilitates the discovery of novel pharmacologically active agents and toxic substances without animal use.
