Summary
Peloruside A (PelA), a novel microtubule-stabilizing agent and potential anti-cancer drug, isolated from the marine sponge Mycale hentscheli, binds to a distinct, non-taxoid binding site on tubulin. Using live-cell confocal microscopy, the effects of PelA on microtubule dynamics were quantified in a human breast adenocarcinoma cell line (MCF7) stably expressing GFP-α-tubulin. Changes in microtubule length were tracked over time in cells treated with PelA concentrations ranging from 3.8–100 nM. As with other microtubule-targeting drugs like paclitaxel and epothilone B, microtubule dynamics were suppressed in a concentration-dependent manner. At the PelA IC50 concentrations for cell proliferation (3.8 nM) and G2/M block (25 nM), PelA inhibited dynamicity by 23% and 45%, respectively. At 25 nM PelA, effects included a 24% and 41% reduction in average growth rate and growth length, respectively. Additionally, the total time spent in pause increased by 53% and coincided with a 36% reduction in the average amount of time spent growing. Rescue and catastrophe frequencies were not significantly affected by PelA, except for length-based catastrophe (67% increase). The results provide further insight into PelA’s unique mode of stabilization and contribute to our understanding of how microtubule-targeting agents exert their anti-mitotic effects.
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Abbreviations
- EpoB:
-
epothilone B
- MSA:
-
microtubule stabilizing agent
- MTA:
-
microtubule targeting agent
- PelA:
-
peloruside A
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Acknowledgments
This work was supported by grants from the Cancer Society of New Zealand and Victoria University of Wellington and a postgraduate scholarship from the Genesis Oncology Trust to A. Chan. We thank Daniel Logan and Cameron Jack for their assistance in setting up the necessary software for measuring microtubule dynamics, and Sushila Pillai for her invaluable assistance using the confocal microscope. We also thank Paraskevi Giannakakou for supplying the GFP-tubulin transfected cells and her constructive feedback regarding the manuscript.
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Chan, A., Andreae, P.M., Northcote, P.T. et al. Peloruside A inhibits microtubule dynamics in a breast cancer cell line MCF7. Invest New Drugs 29, 615–626 (2011). https://doi.org/10.1007/s10637-010-9398-2
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DOI: https://doi.org/10.1007/s10637-010-9398-2