Alpha-phellandrene-induced DNA damage and affect DNA repair protein expression in WEHI-3 murine leukemia cells in vitro

Environ Toxicol. 2015 Nov;30(11):1322-30. doi: 10.1002/tox.22003. Epub 2014 May 24.

Abstract

Although there are few reports regarding α-phellandrene (α-PA), a natural compound from Schinus molle L. essential oil, there is no report to show that α-PA induced DNA damage and affected DNA repair associated protein expression. Herein, we investigated the effects of α-PA on DNA damage and repair associated protein expression in murine leukemia cells. Flow cytometric assay was used to measure the effects of α-PA on total cell viability and the results indicated that α-PA induced cell death. Comet assay and 4,6-diamidino-2-phenylindole dihydrochloride staining were used for measuring DNA damage and condensation, respectively, and the results indicated that α-PA induced DNA damage and condensation in a concentration-dependent manner. DNA gel electrophoresis was used to examine the DNA damage and the results showed that α-PA induced DNA damage in WEHI-3 cells. Western blotting assay was used to measure the changes of DNA damage and repair associated protein expression and the results indicated that α-PA increased p-p53, p-H2A.X, 14-3-3-σ, and MDC1 protein expression but inhibited the protein of p53, MGMT, DNA-PK, and BRCA-1.

Keywords: DNA damage and repair; WEHI-3 leukemia cells; α-phellandrene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anacardiaceae / chemistry
  • Animals
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Comet Assay
  • Cyclohexane Monoterpenes
  • DNA Damage*
  • DNA Repair*
  • DNA-Activated Protein Kinase / genetics
  • DNA-Binding Proteins / genetics*
  • Dose-Response Relationship, Drug
  • Gene Expression / drug effects
  • Mice
  • Microscopy, Confocal
  • Monoterpenes / isolation & purification
  • Monoterpenes / pharmacology*
  • Protein Biosynthesis

Substances

  • Cyclohexane Monoterpenes
  • DNA-Binding Proteins
  • Monoterpenes
  • alpha phellandrene
  • DNA-Activated Protein Kinase