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    Autophagy Inhibition Enhances the Mitochondrial-Mediated Apoptosis Induced by Mangrove (Avicennia marina) Extract in Human Breast Cancer Cells

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    Type
    Article
    Authors
    Esau, Luke
    Sagar, Sunil
    Bajic, Vladimir B. cc
    Kaur, Mandeep
    KAUST Department
    Computational Bioscience Research Center (CBRC)
    Technology Transfer
    Applied Mathematics and Computational Science Program
    Computer, Electrical and Mathematical Sciences and Engineering (CEMSE) Division
    Date
    2015-01-10
    Permanent link to this record
    http://hdl.handle.net/10754/550089
    
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    Abstract
    Aims: Avicennia marina (AM) is a widely distributed mangrove plant that has been used in traditional medicine for centuries for the treatment of a number of diseases. The objective of the present study was to evaluate the leaf ethyl acetate extract of AM for its cytotoxic and apoptotic potential along with in-depth investigations of its mechanism of action in breast cancer MCF-7 cells. Study Design: The ethyl acetate extract of leaves and stems of AM was tested against estrogen positive breast cancer cell line MCF-7 using various assays. Place and Duration of Study: The study was carried out at King Abdullah University of Science and Technology, Thuwal, Saudi Arabia, from July 2013-June 2014. Methodology: Dose- and time-dependent growth inhibition of cancer cells was measured using MTT assay. The mechanisms of apoptosis induction were determined using various assays: phosphatidylserine exposure, caspase-3/7 activation, mitochondrial membrane potential disruption, reactive oxygen species (ROS) production, cell cycle analysis, autophagy, and protein expression using western blotting. The modulation of apoptotic genes (p53, Mdm2, NF-kB, Bad, Bax, Bcl-2 and Casp7) was also determined using real time PCR. Results: The AM extract inhibited breast cancer cell growth and induced apoptosis in a concentration dependent manner. We demonstrated a non-classical mode of apoptosis induction in MCF-7 cells by AM extract, where ROS production altered the mitochondrial membrane potential to induce apoptosis. Breast cancer cells treated with 200 µg/ml concentration of AM extract showed increased ROS production and disrupted MMP but no PARP-1 cleavage and a marked decrease in Caspase-7 protein levels (24 and 48 h) were detected. A significant amount of autophagy was also observed at the same concentration. However, treatment of MCF-7 cells with 200 µg/ml of AM extract along with the inhibition of autophagy by chloroquine, significantly increased the apoptosis from 20% to 45%. Conclusion: Our data provide evidence that AM extract triggers ROS-mediated autophagy as well as caspase-independent apoptosis. The results also strengthen the view that concurrent targeting of apoptotic and autophagic pathways may provide effective therapeutic strategy against cancer.
    Citation
    Autophagy Inhibition Enhances the Mitochondrial-Mediated Apoptosis Induced by Mangrove (Avicennia marina) Extract in Human Breast Cancer Cells 2015, 5 (3):304 European Journal of Medicinal Plants
    Publisher
    Sciencedomain International
    Journal
    European Journal of Medicinal Plants
    DOI
    10.9734/EJMP/2015/14181
    Additional Links
    http://www.sciencedomain.org/abstract.php?iid=791&id=13&aid=7245
    ae974a485f413a2113503eed53cd6c53
    10.9734/EJMP/2015/14181
    Scopus Count
    Collections
    Articles; Applied Mathematics and Computational Science Program; Computational Bioscience Research Center (CBRC); Computer, Electrical and Mathematical Sciences and Engineering (CEMSE) Division

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