Flavokawain A induces apoptosis in MCF-7 and MDA-MB231 and inhibits the metastatic process in vitro

PLoS One. 2014 Oct 6;9(10):e105244. doi: 10.1371/journal.pone.0105244. eCollection 2014.

Abstract

Introduction: The kava-kava plant (Piper methsyticum) is traditionally known as the pacific elixir by the pacific islanders for its role in a wide range of biological activities. The extract of the roots of this plant contains a variety of interesting molecules including Flavokawain A and this molecule is known to have anti-cancer properties. Breast cancer is still one of the leading diagnosed cancers in women today. The metastatic process is also very pertinent in the progression of tumorigenesis.

Methods: MCF-7 and MDA-MB231 cells were treated with several concentrations of FKA. The apoptotic analysis was done through the MTT assay, BrdU assay, Annexin V analysis, cell cycle analysis, JC-1 mitochondrial dye, AO/PI dual staining, caspase 8/9 fluorometric assay, quantitative real time PCR and western blot. For the metastatic assays, the in vitro scratch assay, trans-well migration/invasion assay, HUVEC tube formation assay, ex vivo rat aortic ring assay, quantitative real time PCR and western blot were employed.

Results: We have investigated the effects of FKA on the apoptotic and metastatic process in two breast cancer cell lines. FKA induces apoptosis in both MCF-7 and MDA-MB231 in a dose dependent manner through the intrinsic mitochondrial pathway. Additionally, FKA selectively induces a G2/M arrest in the cell cycle machinery of MDA-MB231 and G1 arrest in MCF-7. This suggests that FKA's anti-cancer activity is dependent on the p53 status. Moreover, FKA also halted the migration and invasion process in MDA-MB231. The similar effects can be seen in the inhibition of the angiogenesis process as well.

Conclusions: FKA managed to induce apoptosis and inhibit the metastatic process in two breast cancer cell lines, in vitro. Overall, FKA may serve as a promising candidate in the search of a new anti-cancer drug especially in halting the metastatic process but further in vivo evidence is needed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Apoptosis / genetics
  • Blotting, Western
  • Breast Neoplasms / blood supply
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology*
  • Caspase 8 / metabolism
  • Caspase 9 / metabolism
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Chalcone / analogs & derivatives*
  • Chalcone / chemical synthesis
  • Chalcone / pharmacology
  • Chalcone / therapeutic use
  • Female
  • G2 Phase / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Humans
  • Inhibitory Concentration 50
  • MCF-7 Cells
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Mitosis / drug effects
  • Neoplasm Invasiveness
  • Neoplasm Metastasis / pathology
  • Neoplasm Metastasis / prevention & control*
  • Neovascularization, Pathologic / drug therapy
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction

Substances

  • flavokawain A
  • Chalcone
  • Caspase 8
  • Caspase 9

Grant support

This study was supported by Fundamental Research Grant Scheme (FRGS/1/2012/SG05/UPM/02/5) from the Ministry of Education, Government of Malaysia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.