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    2020-08-12


    According to the American Cancer Society, a biopsy is usually used to achieve certain tests to determine the speed of cancer growth and find the successful treatment for that, and therefore, the kinetics model with simple formulas and logical precision may have determined the size of a tumor in a function of time. For the untreated breast cancer
    Fig. 7. Cell cycle distribution of the MCF-7 cells treated with IL-GFE at corresponding IC50 in a time-dependent manner.
    MCF-7 cells growth, the first order linear equation is presented as fol-lows:
    After treating MCF7 cells with IL-GFE, the first order linear equation become:
    Additionally, when treating MCF-7 cells with Taxol, the first-order linear equation can be expressed as:
    However, to underestimate the effectiveness of the treatment, the 
    tumor growth characteristic can be neglected (Mehrara et al., 2007). Thus, the final kinetic growth model in response to IL-GFE treatment gives the following equation:
    Moreover, the kinetics’ final growth model after treating with Taxol gives the equation as:
    Where, log10Nf = is the final volume of a tumor after treatment, log10N0 = is the initial volume of a tumor and t = is the duration of treatment (tf-to).
    The specific growth rate (μ) was then used to achieve growth
    kinetics model based on the fourth equation. This model allows to identify the tumor size at an early stage (less than 1 cc) and predict interpolation and extrapolation of sizes at any time points. It appears from the kinetics model constants that IL-GFE treatment has inhibited MCF-7 cells growth. Cancer is known as a disease of the 70132-50-2 dysfunction. The efficient anticancer drug can block the cell cycle progression in cancer cells (Mantena et al., 2006). To investigate whether IL-GFE induced growth inhibition by the cell cycle arrest, MCF-7 cell cycle distribution was examined after IL-GFE-treatment using flow cytometry analysis. The current study demonstrated that IL-GFE arrests the MCF-7-cell cycle at the growth-static G1 phase which ex-plains the anti-proliferation of MCF7 by IL-GFE.
    Furthermore, the results of the flow cytometry analysis using Annexin V/PI staining showed that GFE has remarkedly induced apoptosis in breast cancer cells. This finding was confirmed by Moghadamtousi et al. (2014) and Chamcheu et al. (2018) when treated human HCT-116 and HT-29 colon cancer, and non-melanoma skin cancer NMSC cell lines with ethyl acetate extract of Annona muricata leaves. Moreover, Fig. 8 indicates the appearance of the necrotic cells in a time-dependent manner which is in close agreement with the results of Torres et al. (2012), after treating pancreatic cancer PC cells with Graviola extract. They observed that Graviola extract inhibited cellular metabolism by inhibiting multiple signaling pathways that regulate metabolism, cell cycle and metastatic properties in pancreatic cancer cells. Thus, both mechanisms either apoptosis and necrosis are induced by Graviola fruit ionic liquid extract.
    Intensive phytochemical investigations of the fruit and leaves of Graviola have isolated and identified a great number of acetogenins with a potent biological and pharmacological activities, such as antic-ancer, cytotoxicity, and apoptotic on human cancer cells (Gajalakshmi 
    et al., 2012; Moghadamtousi et al., 2014). Hence, these studies revealed that most acetogenins act as a DNA topoisomerase I poison, induced apoptotic cell death at a Bax and caspase 3 related pathways, arrested cancer cells at the G1 phase and inhibited NADH-ubiquinone oxidor-eductase of the complex I mitochondrial (Rieser et al., 1991). It can be proposed that the synergistic effects of phytochemicals present in the extracts of Graviola fruit including acetogenins may be responsible for the potential chemotherapy agents of Graviola. These acetogenins found in a large quantity in Graviola extracts may act in synergy and might be responsible for their antiproliferation activity. Thus, further studies on the pure isolated compounds from Graviola fruit through bioassay-guided approach are still required to undoubtedly explore the potent active compounds responsible for the antiproliferative activity of the fruit.
    5. Conclusion
    In summary, IL-GFE exhibit antiproliferative effects against MCF-7 breast cancer cell lines by inducing loss of cell viability via apoptosis, morphology changes, and cell cycle arrest at the G0/G1 phase. This inhibition was selective to the growth of MCF-7 cells without any effect on nontumorigenic VERO cells, suggesting that IL-GFE possesses se-lective antitumor properties toward cancer cells. It also showed their potentiality as an anticancer agent when compared with Taxol as a positive control. Moreover, IL-GFE treatment has inhibited breast cancer MCF-7 cell growth by reducing the number of cell generations and increasing the doubling time compared with the control. These data suggest that the IL-GFE can be developed as a novel mechanism-based supplement agent for the cure and prevention of breast cancer. Further investigations are necessary to elucidate the mechanisms underlying