title: Antiproliferative property and apoptotic effect of xanthorrhizol on MDA-MB-231 breast cancer cells.
abstract: Xanthorrhizol is a natural sesquiterpenoid compound isolated from the rhizome of Curcuma xanthorrhizza Roxb (Zingerberaceae). Recent studies of xanthorrhizol in cell cultures strongly support the role of xanthorrhizol as an antiproliferative agent. In our study, we tested the antiproliferative effect of xanthorrhizol using different breast cancer cell lines. The invasive breast cancer cell line, MDA-MB-231, was then selected for further investigations. Treatment with xanthorrhizol caused 50% growth inhibition on MDA-MB-231 cells at 8.67 +/- 0.79 microg/ml as determined by sulforhodamine B (SRB) assay. Hoechst 33258 nuclear staining assay showed the rate of apoptosis of MDA-MB-231 cells to increase in response to xanthorrhizol treatment. Immunofluorescence staining using antibody MitoCapture and fluorescein isothiocyanate (FITC)-labeled cytochrome c revealed the possibility of altered mitochondrial transmembrane potential and the release of cytochrome c respectively. This was further confirmed by Western-blotting, where cytochrome c was showed to migrate from mitochondrial fraction to the cytosol fraction of treated MDA-MB-231 cells. Caspase activity assay showed the involvement of caspase-3 and caspase-9, but not caspase-6 or caspase-8 in MDA-MB-231 apoptotic cell death. Subsequently, cleavage of PARP-1 protein is suggested. These data suggest treatment with xanthorrhizol modulates MDA-MB-231 cell apoptosis through the mitochondria-mediated pathway subsequent to the disruption of mitochondrial transmembrane potential, release of cytochrome c, activation of caspase-3 and caspase-9, and the modulation of PARP-1 protein.
authors.affiliation: Bioassay Unit, Herbal Medicine Research Center, Institute for Medical Research, Jalan Pahang, Kuala Lumpur, Malaysia. yhcheah@gmail.com
authors.forename: Yew Hoong
authors.initials: YH
authors.lastname: Cheah
authors.forename: Fariza Juliana
authors.initials: FJ
authors.lastname: Nordin
authors.forename: Thiam Tsui
authors.initials: TT
authors.lastname: Tee
authors.forename: Hawariah Lope Pihie
authors.initials: HL
authors.lastname: Azimahtol
authors.forename: Noor Rain
authors.initials: NR
authors.lastname: Abdullah
authors.forename: Zakiah
authors.initials: Z
authors.lastname: Ismail
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole / Q000737:chemistry; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D020169:Caspases / Q000378:metabolism; D002417:Cattle; D048708:Cell Growth Processes / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D010636:Phenols / Q000494:pharmacology*; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods; D014709:Vero Cells
chemical_list: D007527:Isoenzymes; D010636:Phenols; C120248:xanthorrhizol; D045304:Cytochromes c; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases; D006690:Bisbenzimidazole