Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Acacetin-induced Apoptosis of Human Breast Cancer MCF-7 Cells Involves Caspase Cascade, Mitochondria-mediated Death Signaling and SAPK/JNK1/2-c-Jun Activation

  • Shim, Hye-Young (Department of Advanced Technology Fusion and Bio/Molecular Informatics Center, Konkuk University) ;
  • Park, Jong-Hwa (Department of Advanced Technology Fusion and Bio/Molecular Informatics Center, Konkuk University) ;
  • Paik, Hyun-Dong (Division of Animal Life Science and Bio/Molecular Informatics Center, Konkuk University) ;
  • Nah, Seung-Yeol (Department of Physiology, Collage of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Kim, Darrick S.H.L. (CurXceL Corporation, The Business and Technology Center) ;
  • Han, Ye Sun (Department of Advanced Technology Fusion and Bio/Molecular Informatics Center, Konkuk University)
  • Received : 2007.02.07
  • Accepted : 2007.05.02
  • Published : 2007.08.31
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Abstract

The mechanism of acacetin-induced apoptosis of human breast cancer MCF-7 cells was investigated. Acacetin caused 50% growth inhibition ($IC_{50}$) of MCF-7 cells at $26.4{\pm}0.7{\mu}M$ over 24 h in the MTT assay. Apoptosis was characterized by DNA fragmentation and an increase of sub-G1 cells and involved activation of caspase-7 and PARP (poly-ADP-ribose polymerase). Maximum caspase 7 activity was observed with $100{\mu}M$ acacetin for 24 h. Caspase 8 and 9 activation cascades mediated the activation of caspase 7. Acacetin caused a reduction of Bcl-2 expression leading to an increase of the Bax:Bcl-2 ratio. It also caused a loss of mitochondrial membrane potential that induced release of cytochrome c and apoptosis inducing factor (AIF) into the cytoplasm, enhancing ROS generation and subsequently resulting in apoptosis. Pretreatment of cells with N-acetylcysteine (NAC) reduced ROS generation and cell growth inhibition, and pretreatment with NAC or a caspase 8 inhibitor (Z-IETD-FMK) inhibited the acacetin-induced loss of mitochondrial membrane potential and release of cytochrome c and AIF. Stress-activated protein kinase/c-Jun $NH_4$-terminal kinase 1/2 (SAPK/JNK1/2) and c-Jun were activated by acacetin but extracellular-regulated kinase 1/2 (Erk1/2) nor p38 mitogen-activated protein kinase (MAPK) were not. Our results show that acacetin-induced apoptosis of MCF-7 cells is mediated by caspase activation cascades, ROS generation, mitochondria-mediated cell death signaling and the SAPK/JNK1/2-c-Jun signaling pathway, activated by acacetin-induced ROS generation.

Keywords

Acknowledgement

Supported by : Korea Research Foundation, Korean Ministry of Science and Technology

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