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  • br Djabir Daddiouaissaa Azura Amidb Nassereldeen A Kabbashia

    2020-08-12


    Djabir Daddiouaissaa, Azura Amidb,∗, Nassereldeen A. Kabbashia, Fazia A.A. Fuada, AhmedA.M. Elnoura, Mohamad A.K.M. S. Epandyc
    a Biotechnology Engineering, Kulliyyah of Engineering, International Islamic University, Malaysia (IIUM), P. O. Box 10, Gombak, 50728, Kuala Lumpur, Malaysia
    b International Institute for Halal Research and Training (INHART), Level 3, KICT Building, International Islamic University Malaysia (IIUM), Jalan Gombak, 53100, Kuala Lumpur, Malaysia
    c Adikafirdaus Resources, Lot 24, Jalan Klebang Selatan, 2/5 Kampung Tersusun, Batu 6 Klebang Selatan, 31200, Ipoh, Perak, Malaysia
    Keywords:
    Breast cancer
    Graviola
    Ionic liquid
    Flow cytometry
    Apoptosis 
    Ethnopharmacological relevance: Medicinal plants have been used for ages by indigenous communities around the world to help humankind sustain its health. Graviola (Annona muricata), also called soursop, is a member of the Annonaceae family and is an evergreen plant that is generally distributed in tropical and subtropical areas of the world. Graviola tree has a long history of traditional use due to its therapeutic potential including anti-in-flammatory, antimicrobial, antioxidant, insecticide and cytotoxic to tumor cells.
    Aim of the study: This study aimed to investigate the in vitro antiproliferative effects and apoptotic events of the ionic liquid extract of Graviola fruit (IL-GFE) on MCF-7 breast cancer cells and their cytokinetics behaviour to observe their potential as a therapeutic alternative in cancer treatment.
    Materials and methods: The cell viability assay of the extract was measured using tetrazolium bromide (MTT assay) to observe the effects of Graviola fruit extract. Then the cytokinetics behaviour of MCF-7 cells treated with IL-GFE is observed by plotting the growth curve of the cells. Additionally, the Hygromycin B distribution and apoptosis mechanism of IL-GFE action on MCF-7 cancer cells were observed by flow cytometry.
    Results: IL-GFE exhibited anti-proliferative activity on MCF-7 with the IC50 value of 4.75 μg/mL, compared to Taxol with an IC50 value of 0.99 μg/mL. IL- GFE also reduced the number of cell generations from 3.71 to 1.67 generations compared to 2.18 generations when treated with Taxol. Furthermore, the anti-proliferative activities were verified when the growth rate was decreased dynamically from 0.0077 h to 1 to 0.0035 h-1. Observation of the IL-GFE-treated MCF-7 under microscope demonstrated detachment of cells and loss of density. The growth inhibition of the cells by extracts was associated with cell cycle arrest at the G0/G1 phase, and phosphati-dylserine externalisation confirms the anti-proliferation through apoptosis.
    Conclusions: ionic liquid Graviola fruit extract affect the cytokinetics behaviour of MCF-7 cells by reducing cell viability, induce apoptosis and cell cycle arrest at the G0/G1 phase.
    1. Introduction
    Fruit consumption is highly recommended to maintain human health (Halliwell, 2012). Graviola (Annona muricata L.) is also called soursop because of the sour and sweet flavour of its great fruit (Patel and Patel, 2016). Graviola is native to the tropical zones, South America and Africa but is currently widely cultivated in the tropical and sub-tropical zones around the world. Recently, Graviola fruit is widely used to make candies, syrups and beverages (Ioannis et al., 2015). Ethno-botanical studies reported that all parts of Graviola tree are used in an alternative medicine, especially the fruit which is taken to treat fever
    ∗ Corresponding author. E-mail address: [email protected] (A. Amid). 
    (Taylor, 2002), eliminate worms and parasites (Yajid et al., 2018), in-crease mother's milk (Badrie and Schauss, 2010), as well as hyperten-sion (Samuel et al., 2010), liver and renal infections (Coe, 2008). The popular use of Graviola as an anticancer treatment reported ethnobo-tanically may be related to the reports of its selective cytotoxicity (George et al., 2012; Monigatti et al., 2013). This activity is considered selective as some reported in vitro studies have shown that the Graviola extract was more toxic to cancer cells than normal cells (Betancur-Galvis et al., 1999; Dai et al., 2011; Gavamukulya et al., 2014).
    The major benefit of this fruit is the active compounds known as annonaceous acetogenins which are thought to be potent anticancer
    D. Daddiouaissa, et al.
    toward several cancer cell lines including breast cancer as highlighted recently by Magadi et al. (2015) and Daddiouaissa & Amid (2018). At present, the attention is being focused on anticancer properties of Graviola acetogenins which are the primary bioactive compounds found in the fruit (Sun et al., 2017).