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Identification of small molecules inducing apoptosis by cell-based assay using fission yeast deletion mutants

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Summary

The cell-based assay using yeast deletion mutants has been recognized as an efficient analysis to discover therapeutic compounds and reveal their mode of action. In this study, S. pombe deletion mutants-based HTS screening was carried out to identify potential anti-cancer agents. The NCI chemical library of 5700 compounds was screened using kit strains, which consisted of S. pombe mutants harboring deletions in genes involved in DNA repair and mitotic control. During the screening, we identified 40 compounds conferring growth inhibition of S. pombe. Their anti-tumorigenic properties were examined by phenotypic effect on S. pombe, flow cytometry and apoptosis analysis of human cancer. Here, we report hit compounds inducing apoptosis for development of anti-cancer agents suggesting that S. pombe deletion mutants are useful in identifying potential anti-cancer agents in human cancer therapeutics.

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Acknowledgments

This work was supported, in part, by the Grant KRIBB Initiative from the Korea Research Council of Fundamental Science and Technology, a grant from the 21st Century Frontier for Functional Analysis of the Human Genome.

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Authors and Affiliations

  1. Functional Genomics, KRIBB, Yusong, Daejeon, 305-806, South Korea

    Kyung-Sook Chung, Nam-Hui Yim, Shin-Jung Choi, Kyung-Sun Hur, Kwang-Lae Hoe, Dong-Uk Kim, Hyang-Sook Yoo & Misun Won

  2. College of Pharmacy, Chungnam National University, Daejon, 305-764, South Korea

    Nam-Hui Yim & Ki-Hwan Bae

  3. Department of Food Science and Technology, Chungnam National University, Daejon, South Korea

    Seung-Hee Lee & Kyung-Bin Song

  4. Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., Seattle, WA, 98109, USA

    Sondra Goehle & Julian Simon

  5. Department of Biology, University of Korea, Seoul, South Korea

    Hyung-Bae Kim

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  1. Kyung-Sook Chung
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  2. Nam-Hui Yim
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  13. Julian Simon
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  14. Misun Won
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Corresponding author

Correspondence to Julian Simon.

Additional information

Kyung-Sook Chung, Nam-Hui Yim and Seung-Hee Lee were equally contributed.

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Table 1

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Fig. 1

Determination of IC50 by [3H]-thymidine incorporation assay. IC50 was measured with a modified [3H]-thymidine incorporation assay as described previously [18, 19]. To test their apoptotic effect on cancer cells, A549 (lung cancer), MCF7JK (breast cancer), Hct-116 (colon cancer), HeLa cell (cervic cancer), DU-145 (prostate cancer), IGROV1 (ovary cancer), Ovcar3 (ovary cancer), Ovcar8 (ovary cancer), SKOV3 (ovary cancer), MDA-MB 435 (breast cancer), NCI H460 (colon cancer), Daudi (Lymphoma), HFF, human foreskin cell and NCIH460 cells (colon cancer) were used. A. 1, NSC 15627; 2, NSC 280891; 3, NSC 280892; 4, NSC 126188. B. 1, NSC 337859; 2, NSC 280891; 3, NSC 280892. C. 1, NSC 658139; 2, NSC 87538; 3, NSC 205357; 4, NSC 10120.(PPT 81.5 KB)

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Chung, KS., Yim, NH., Lee, SH. et al. Identification of small molecules inducing apoptosis by cell-based assay using fission yeast deletion mutants. Invest New Drugs 26, 299–307 (2008). https://doi.org/10.1007/s10637-007-9100-5

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