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Serotonin type-3 receptor antagonists selectively kill melanoma cells through classical apoptosis, microtubule depolymerisation, ERK activation, and NF-κB downregulation

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Abstract

Malignant melanoma is a highly metastatic tumour, resistant to treatment. Serotonin type-3 (5-HT3) receptor antagonists, such as tropisetron and ondansetron, are well-tolerated antiemetic drugs commonly used to prevent nausea caused by chemotherapy or radiotherapy. We investigated the anticancer effects of these drugs on melanoma cancer cell lines WM-266–4 and B16F10 with or without paclitaxel. We constructed IC50 curves and performed Chou–Talalay analysis, using data obtained with the MTT assay. Flow cytometry and fluorescent microscopy were used to examine characteristics of the cell cycle, cell death and cytoskeleton changes. Protein levels and activation were analysed by western blotting and molecular docking studies carried out. Data were analysed by one way ANOVA and post hoc testing. Ondansetron and tropisetron showed selective concentration-dependent cytotoxicity in melanoma cell lines WM-266–4 and B16F10. The effect in combination with paclitaxel was synergistic. The drugs did not cause cell cycle arrest but did promote characteristics of classical apoptosis, including accumulation of subG1 DNA, cleaved caspase-3, mitochondrial membrane permeability and phosphatidylserine exposure. As well, the cytosolic calcium level in the melanoma cells was enhanced, phosphorylated ERK1/2 induced and NF-κB inhibited. Finally, the formation of microtubules was shown to be impaired in melanoma cells treated with ondansetron or tropisetron. Docking studies were used to predict that these drugs could bind to the colchicine binding site on the tubulin molecule. Antiemetic drugs, already given in combination with chemotherapy, may enhance the cytotoxic effect of chemotherapy, following successful delivery to the tumour site.

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Data availability

The datasets supporting the conclusions of this article are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Abbreviations

5-HT3:

Serotonin type-3 receptor

TRO:

Tropisetron

OND:

Ondansetron

NHDF:

Normal Human Dermal Fibroblasts

FBS:

Foetal bovine serum

PTX:

Paclitaxel

PBS:

Phosphate-buffered saline

PFA:

Paraformaldehyde

RT:

Room temperature

PI:

Propidium iodide

TMRE:

Tetramethylrhodamine ethyl ester

BSA:

Bovine serum albumin

SD:

Standard deviation

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Acknowledgements

The authors would like to thank Dr Joel Tyndall for his kind help with the docking studies. This work was funded by a University of Otago School of Biomedical Sciences’ Dean’s Bequest Grant and a University of Otago Research Grant.

Funding

This work was funded by a University of Otago School of Biomedical Sciences’ Dean’s Bequest Grant and a University of Otago Research Grant.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand

    Anita Barzegar-fallah, Houman Alimoradi, Jessica L. Dunlop, Elham Torbati & Sarah K. Baird

  2. Department of Biochemistry, University of Otago, Dunedin, New Zealand

    Elham Torbati

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  1. Anita Barzegar-fallah
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  2. Houman Alimoradi
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  3. Jessica L. Dunlop
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  4. Elham Torbati
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  5. Sarah K. Baird
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Contributions

ABF and HA designed and carried out the cell culture-based experimental work for this study, analysed the data, and wrote the initial draft of the manuscript. ET carried out the docking studies and SB obtained the funding, was part of designing the experiments, analysing the data, and finalising the manuscript.

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Correspondence to Sarah K. Baird.

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Barzegar-fallah, A., Alimoradi, H., Dunlop, J.L. et al. Serotonin type-3 receptor antagonists selectively kill melanoma cells through classical apoptosis, microtubule depolymerisation, ERK activation, and NF-κB downregulation. Cell Biol Toxicol 39, 1119–1135 (2023). https://doi.org/10.1007/s10565-021-09667-0

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