title: Induction of necrosis and cell cycle arrest in murine cancer cell lines by Melaleuca alternifolia (tea tree) oil and terpinen-4-ol.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Discipline of Microbiology and Immunology (M502), School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia. srooney@cyllene.uwa.edu.au
authors.forename: S J
authors.initials: SJ
authors.lastname: Greay
authors.forename: D J
authors.initials: DJ
authors.lastname: Ireland
authors.forename: H T
authors.initials: HT
authors.lastname: Kissick
authors.forename: A
authors.initials: A
authors.lastname: Levy
authors.forename: M W
authors.initials: MW
authors.lastname: Beilharz
authors.forename: T V
authors.initials: TV
authors.lastname: Riley
authors.forename: C F
authors.initials: CF
authors.lastname: Carson
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D008546:Melanoma, Experimental / Q000188:drug therapy; D008654:Mesothelioma / Q000188:drug therapy; D051379:Mice; D009336:Necrosis / Q000139:chemically induced; D020947:Tea Tree Oil / Q000627:therapeutic use*; D013729:Terpenes / Q000627:therapeutic use*
chemical_list: D000970:Antineoplastic Agents; D013729:Terpenes; C034019:terpinenol-4; D020947:Tea Tree Oil