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Induction of necrosis and cell cycle arrest in murine cancer cell lines by Melaleuca alternifolia (tea tree) oil and terpinen-4-ol

Abstract

Purpose

To examine the in vitro anticancer activity of Melaleuca alternifolia (tea tree) oil (TTO), and its major active terpene component, terpinen-4-ol, against two aggressive murine tumour cell lines, AE17 mesothelioma and B16 melanoma.

Methods

Effects of TTO and terpinen-4-ol on the cellular viability of two tumour cell lines and fibroblast cells were assessed by MTT assay. Induction of apoptotic and necrotic cell death was visualised by fluorescent microscopy and quantified by flow cytometry. Tumour cell ultrastructural changes were examined by transmission electron microscopy and changes in cell cycle distribution were assessed by flow cytometry, with changes in cellular morphology monitored by video time lapse microscopy.

Results

TTO and terpinen-4-ol significantly inhibited the growth of two murine tumour cell lines in a dose- and time-dependent manner. Interestingly, cytotoxic doses of TTO and terpinen-4-ol were significantly less efficacious against non-tumour fibroblast cells. TTO and terpinen-4-ol induced necrotic cell death coupled with low level apoptotic cell death in both tumour cell lines. This primary necrosis was clarified by video time lapse microscopy and also by transmission electron microscopy which revealed ultrastructural features including cell and organelle swelling following treatment with TTO. In addition, both TTO and terpinen-4-ol induced their inhibitory effect by eliciting G1 cell cycle arrest.

Conclusion

TTO and terpinen-4-ol had significant anti-proliferative activity against two tumour cell lines. Moreover, the identification of primary necrotic cell death and cell cycle arrest of the aggressive tumour cells highlights the potential anticancer activity of TTO and terpinen-4-ol.

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Acknowledgments

This work was supported by a grant (PRJ-002395) from the Rural Industries Research and Development Corporation ACT, Australia and Novasel Australia Pty. Ltd., Mudgeeraba, QLD, Australia. We acknowledge the facilities, scientific and technical assistance of the Australian Microscopy & Microanalysis Research Facility at the Centre for Microscopy, Characterisation & Analysis (CMCA), The University of Western, Australia, a facility funded by The University, State and Commonwealth Governments and in particular Dr Paul Rigby (CMCA) for his help with the video time lapse microscopy. We would like to thank Pierre Filion and Robert Cook from the Electron Microscopy Unit at PathWest Laboratory Medicine WA for their technical advice, the use of reagents and equipment and helpful discussions.

Author information

Correspondence to S. J. Greay.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Representative video time lapse microscopy of AE17 control cells over 24 h (WMV 9763 kb)

Representative video time lapse microscopy of AE17 TTO (0.04%) treated cells over 24 h (WMV 9365 kb)

Representative video time lapse microscopy of B16 control cells over 24 h (WMV 9591 kb)

Representative video time lapse microscopy of B16 TTO (0.04%) treated cells over 24 h (WMV 9334 kb)

Supplementary Movie 1

Representative video time lapse microscopy of AE17 control cells over 24 h (WMV 9763 kb)

Supplementary Movie 2

Representative video time lapse microscopy of AE17 TTO (0.04%) treated cells over 24 h (WMV 9365 kb)

Supplementary Movie 3

Representative video time lapse microscopy of B16 control cells over 24 h (WMV 9591 kb)

Supplementary Movie 4

Representative video time lapse microscopy of B16 TTO (0.04%) treated cells over 24 h (WMV 9334 kb)

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Greay, S.J., Ireland, D.J., Kissick, H.T. et al. Induction of necrosis and cell cycle arrest in murine cancer cell lines by Melaleuca alternifolia (tea tree) oil and terpinen-4-ol. Cancer Chemother Pharmacol 65, 877–888 (2010). https://doi.org/10.1007/s00280-009-1093-7

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Keywords

  • Tea tree oil
  • Terpenes
  • Necrosis
  • Apoptosis
  • Cell cycle arrest
  • Anticancer