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Putative mechanisms of antitumor activity of cyano-substituted heteroaryles in HeLa cells

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Summary

Six recently synthesized cyano-substituted heteroaryles, which do not bind to DNA but are highly cytotoxic against the human tumor cell line HeLa, were analyzed for their antitumor mechanisms of action (MOA). They did not interfere with the expression of human papillomavirus oncogenes integrated in the HeLa cell genome, but they did induce strong G1 arrest and result in the activation of caspase-3 and apoptosis. A computational analysis was performed that compared the antiproliferative activities of our compounds in 13 different tumor cell lines with those of compounds listed in the National Cancer Institute database. The results indicate that interference with cytoskeletal function and inhibition of mitosis are the likely antitumor MOA. Furthermore, a second in silico investigation revealed that the tumor cells that are sensitive to the cyano-substituted compounds show differences in their expression of locomotion genes compared with that of insensitive cell lines, thus corroborating the involvement of the cytoskeleton. This MOA was also confirmed experimentally: the cyano-substituted heteroaryles disrupted the actin and the tubulin networks in HeLa cells and inhibited cellular migration. However, further analysis indicated that multiple MOA may exist that depend on the position of the cyano-group; while cyano-substituted naphthiophene reduced the expression of cytoskeletal proteins, cyano-substituted thieno-thiophene-carboxanilide inhibited the formation of cellular reactive oxygen species.

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Acknowledgments

We thank Dr. Jaganjac for assistance with measurements of intracellular ROS, and to Dr. Čimbora-Zovko who kindly provided antibodies for cytoskeletal-study. We also thank to Dr. Ragland for helpful comments.

This study was supported by grants from the Ministry of Science, Education and Sports of the Republic of Croatia to MK (098-0982464-2514) and to TŠ (098-0000000-3168) and from Ministero dell’Istruzione, dell’Università e della Ricerca, Italy (PRIN 2008) to DL.

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Authors and Affiliations

  1. Division of Molecular Medicine, Ruđer Bošković Institute, Bijenička 54, HR-10000, Zagreb, Croatia

    Katja Ester, Kristina Majsec, Marko Marjanović & Marijeta Kralj

  2. Division of Electronics, Ruđer Bošković Institute, Bijenička 54, HR-10000, Zagreb, Croatia

    Fran Supek & Tomislav Šmuc

  3. Department of Clinical and Biological Sciences, S. Luigi Gonzaga Medical School, University of Turin, 10043, Orbassano, Turin, Italy

    David Lembo & Manuela Donalisio

  4. Department of Organic Chemistry, Faculty of Chemical Engineering and Technology, University of Zagreb, Marulićev trg 18, HR-10000, Zagreb, Croatia

    Ivana Jarak & Grace Karminski-Zamola

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  1. Katja Ester
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  2. Fran Supek
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Correspondence to Katja Ester.

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Ester, K., Supek, F., Majsec, K. et al. Putative mechanisms of antitumor activity of cyano-substituted heteroaryles in HeLa cells. Invest New Drugs 30, 450–467 (2012). https://doi.org/10.1007/s10637-010-9571-7

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