J Cancer Prev.
2015 Dec;20(4):241-249. 10.15430/JCP.2015.20.4.241.
Anti-cancer Activity of Osmanthus matsumuranus Extract by Inducing G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma Hep G2 Cells
- Affiliations
-
- 1Blue-Bio Industry Regional Innovation Center, Dong-Eui University, Busan, Korea. bwkim@deu.ac.kr
- 2Department of Life Science and Biotechnology, College of Natural Sciences and Human Ecology, Dong-Eui University, Busan, Korea.
- 3Department of Biochemistry, College of Korean Medicine, Dong-Eui University, Busan, Korea.
- 4Anti-Aging Research Center, Dong-Eui University, Busan, Korea.
- KMID: 2380876
- DOI: http://doi.org/10.15430/JCP.2015.20.4.241
Abstract
- BACKGROUND
Osmanthus matsumuranus, a species of Oleaceae, is found in East Asia and Southeast Asia. The bioactivities of O. matsumuranus have not yet been fully understood. Here, we studied on the molecular mechanisms underlying anti-cancer effect of ethanol extract of O. matsumuranus (EEOM).
METHODS
Inhibitory effect of EEOM on cell growth and proliferation was determined by WST assay in various cancer cells. To investigate the mechanisms of EEOM-mediated cytotoxicity, HepG2 cells were treated with various concentration of EEOM and analyzed the cell cycle arrest and apoptosis induction by flow cytometry, Western blot analysis, 4,6-diamidino-2-phenylindole (DAPI) staining and DNA fragmentation.
RESULTS
EEOM showed the cytotoxic activities in a dose-dependent manner in various cancer cell lines but not in normal cells, and HepG2 cells were most susceptible to EEOM-induced cytotoxicity. EEOM induced G2/M arrest in HepG2 cells associated with decreased expression of cyclin-dependent kinase 1 (CDK1), cyclin A and cylcin B, and increased expression of phospho-checkpoint kinase 2, p53 and CDK inhibitor p21. Immunofluorescence staining showed that EEOM-treated HepG2 increased doublet nuclei and condensed actin, resulting in cell rounding. Furthermore, EEOM-mediated apoptosis was determined by Annexin V staining, chromatin condensation and DNA fragmentation. EEOM caused upregulation of FAS and Bax, activation of caspase-3, -8, -9, and fragmentation of poly ADP ribose polymerase.
CONCLUSIONS
These results suggest that EEOM efficiently inhibits proliferation of HepG2 cells by inducing both G2/M arrest and apoptosis via intrinsic and extrinsic pathways, and EEOM may be used as a cancer chemopreventive agent in the food or nutraceutical industry.
MeSH Terms
-
Actins
Annexin A5
Apoptosis*
Asia, Southeastern
Blotting, Western
Carcinoma, Hepatocellular*
Caspase 3
CDC2 Protein Kinase
Cell Cycle Checkpoints
Cell Line
Chromatin
Cyclin A
Dietary Supplements
DNA Fragmentation
Ethanol
Far East
Flow Cytometry
Fluorescent Antibody Technique
Hep G2 Cells*
Humans*
Oleaceae
Phosphotransferases
Poly(ADP-ribose) Polymerases
Up-Regulation
Actins
Annexin A5
CDC2 Protein Kinase
Caspase 3
Chromatin
Cyclin A
Ethanol
Phosphotransferases
Poly(ADP-ribose) Polymerases
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