Yeast caspase-dependent apoptosis in Saccharomyces cerevisiae BY4742 induced by antifungal and potential antitumor agent
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ORIGINAL PAPER
Yeast caspase‑dependent apoptosis in Saccharomyces cerevisiae BY4742 induced by antifungal and potential antitumor agent clotrimazole Berna Kavakçıoğlu1 · Leman Tarhan1
Received: 23 April 2017 / Revised: 25 July 2017 / Accepted: 7 August 2017 © Springer-Verlag GmbH Germany 2017
Abstract Clotrimazole is an antifungal medication commonly used in the treatment of fungal infections. There is also promising research on using clotrimazole against other diseases such as malaria, beriberi, tineapedis and cancer. It was aimed to investigate the apoptotic phenotype in Saccharomyces cerevisiae induced by clotrimazole. The exposure of S. cerevisiae to 10 µM clotrimazole for 3, 6 and 9 h caused to decrease in cell viability by 24.82 ± 0.81, 56.00 ± 1.54 and 77.59 ± 0.53%, respectively. It was shown by Annexin V–PI assay that 110 µM clotrimazole treatment caused to death by 35.5 ± 2.48% apoptotic and only 13.1 ± 0.08% necrotic pathway within 30 min. The occurrence of DNA strand breaks and condensation could be visualised by the TUNEL and DAPI stainings, respectively. Yeast caspase activity was induced 12.34 ± 0.71-fold after 110 µM clotrimazole treatment for 30 min compared to the control. The dependency of clotrimazole-induced apoptosis to caspase was also shown using Δyca1 mutant. Keywords Saccharomyces cerevisiae · Clotrimazole · Viability · Apoptosis · Yeast caspase
Communicated by Erko Stackebrandt. * Leman Tarhan [email protected] Berna Kavakçıoğlu [email protected] 1
Department of Chemistry, Faculty of Science, University of Dokuz Eylul, Tinaztepe Campus, 35160 Buca, Izmir, Turkey
Introduction Clotrimazole (CLT), a synthetic and lipophilic imidazole derivative, is primarily used to treat infections caused by various fungi. It is known for a long time that Candida spp., Vulvovaginal candidiasis, Trichophyton spp., Microsporum spp. and Malazzesia furfur are the strains against which drug is most active (Sawyer et al. 1975). As all azole-type antimycotic drugs, CLT interferes with the biosynthesis of ergosterol, a major component of the fungal cytoplasmic membrane. Specifically, azoles inhibit the microsomal cytochrome P450 (CYP450)dependent event 14-α-lanosterol demethylation, which is a vital step in ergosterol biosynthesis by fungi (Hitchcock et al. 1990). The resultant depletion of ergosterol and its replacement with the aberrant sterol species, 14-α-methylsterol, perturb normal membrane permeability and fluidity (Crowley and Gallagher 2014). While CYP450 inhibition is accepted to primarily account for the antimycotic properties of CLT, this antifungal azole derivative was also recognized as an effective calmodulin antagonist by 1990s (Hegemann et al. 1993; Mac Neil et al. 1993). Based on the knowledge that calmodulin antagonists cause to the defects in the regulation of glycolytic pathway, primary energy source of cancerous cells, researches have been oriented to explore whether CLT also exerts anticancer effect and its possible use in chemotherapy. It was determined that CLT
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