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Sertaconazole induced toxicity in HeLa cells through mitotic arrest and inhibition of microtubule assembly

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Abstract

Econazole, miconazole, and sertaconazole, the structurally related azoles with imidazole moiety, were evaluated for their cytotoxicity and their ability to bind to mammalian tubulin. Our results indicated that sertaconazole and econazole bound to goat brain tubulin with a dissociation constant of 9 and 19 μM respectively, while miconazole did not bind to goat brain tubulin. Econazole, miconazole, and sertaconazole inhibited the proliferation of HeLa cells with an IC50 of 28, 98, and 38 μM respectively with sertaconazole alone inducing a mitotic block in the treated cells. Since sertaconazole bound to goat brain tubulin with higher affinity and blocked the cells at mitosis, we hypothesized that its cytotoxic mechanism might involve inhibition of tubulin and econazole which did not block the cells at mitosis may have additional targets than tubulin. Sertaconazole inhibited the polymerization of tubulin in HeLa cells and the in vitro assembled goat brain tubulin. Competitive tubulin-binding assay using colchicine and computational simulation studies showed that sertaconazole bound closer to the colchicine site and induced the tubulin dimer to adopt a “bent” conformation which is incompetent for the polymerization. Results from RT-PCR analysis of the A549 cells treated with sertaconazole indicated activation of apoptosis. Sertaconazole significantly inhibited the migration of HeLa cells and showed synergistic antiproliferative potential with vinblastine. Collectively, the results suggest that sertaconazole which is already in clinical practice could be useful as a topical chemotherapy agent for the treatment of skin cancers in combination with other systemic anticancer agents.

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All data analyzed during this study are included as Electronic supplementary material (ESM).

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Funding

This study is funded by the NITC and MHRD, Government of India in the form of scholarship to Mr. Jomon Sebastian and Infrastructural facilities to Dr. Rathinasamy K.

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  1. School of Biotechnology, National Institute of Technology Calicut, Calicut, Kerala, 673601, India

    Jomon Sebastian & Krishnan Rathinasamy

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  1. Jomon Sebastian
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  2. Krishnan Rathinasamy
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Contributions

JS performed the experiments, analyzed the data, and wrote the manuscript. KR designed the experiments, provided the resources for the work, critically analyzed the data, and wrote the manuscript. All the authors have read and approved the final version of the manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Krishnan Rathinasamy.

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Sebastian, J., Rathinasamy, K. Sertaconazole induced toxicity in HeLa cells through mitotic arrest and inhibition of microtubule assembly. Naunyn-Schmiedeberg's Arch Pharmacol 394, 1231–1249 (2021). https://doi.org/10.1007/s00210-021-02059-5

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