br Cancer growth model in some
Cancer growth model in some types of cancers can give a good approximation of the volume of cancer Solasodine using specific growth rate (Mehrara et al., 2007). Therefore, to evaluate the eﬀect of IL-GFE on the specific growth of MCF-7, the graph of the log of viable cells number vs. time was plotted, and the regression slope was taken as a specific growth rate (μ). The growth rate of IL-GFE treated cells and taxol were observed to reduce significantly 0.0035 h−1 and 0.012 h−1, respec-tively compared to 0.0077 h−1 in control. r> Table 1 Number of Generations (X), specific growth rate (μ) and doubling time (td) for Untreated MCF-7 Cells (control), MCF-7 Cells Treated with IL-GFE and Taxol.
Cell growth Maximum cells volume (N) (cells/mL) Number of generations (X) Specific growth rate μ (h−1) Doubling time td (h)
Initial cells number (N0) was constant at 2 × 105 cells/mL for all experiments. * Identical superscripts in the same column indicate significant diﬀerence (p ≤ 0.05).
Fig. 6. Representative photographs of A; Untreated MCF-7 cells, and B; MCF-7 cells treated with IL-GFE at a designated time point.
3.3. Cell cycle distribution and apoptosis
Cell cycle progression is well-correlated to the proliferation of the cell. The untreated and the treated MCF-7 cells with IL-GFE were ana-lyzed using RNase A/PI staining in combination with flow cytometry. The results show that IL-GFE treatment at IC50 for 24, 48 and 72 h in-creased the cells in the G0/G1 phase from 59.07% in control into 72.82% in GFE-treated cells for 72 h, while decreased in the S and G2M phase compared with the control. Moreover, cell number at the sub G0 phase of the treated MCF-7 cells increased in a time-dependent manner compared to the control, taking cells at the sub G0 phase as apoptotic cells (Agu et al., 2018); there was a remarkable increase in the number of apoptotic cells starting from 24 h of IL-GFE treatment (Fig. 7).
Next, Annexin-V-FITC assay has been performed to confirm the apoptotic eﬀect of the IL-GFE on MCF-7 cells. Figure (8) shows that the extract has significantly increased the percentage of cells positive to the Annexin-V-FITC staining in range of 0.32% for the control to 14.09% after 72 h of IL-GFE-treatment which indicates apoptosis.
A healthy diet (HD) rich in fruits and vegetables is associated with maintenance of health and reduced risk of diseases including cancers. This is due to the various phytochemicals such as phenols, alkaloids, flavonoids, carotenoids, and vitamins that play an essential role in boosting immunity (Wang et al., 2012). Cancer is treated nowadays using various anticancer drugs such as Taxol, cisplatin, topotecan, Doxorubicin, and etoposide. However, these drugs are known to give severe side eﬀects to the cancer patients due to their ability to kill healthy cells (Magadi et al., 2015). Hence, the search for new antic-ancer drugs with minimal side eﬀects is vital. According to the U.S. National Cancer Institute (NCI), drugs with IC50 values under 20 μg/mL after an incubation time of 48–72 h are considered as a potent cytotoxic substance (Boik, 2001).
Graviola fruit is rich in secondary metabolites such as alkaloids, flavonoids, saponins, and importunately acetogenins which have proved to be a promising anticancer drug. In the present study, the anticancer activity (AC) of ionic liquid-Graviola fruit extract was in-vestigated on adherent breast cancer MCF-7 and standard VERO cell lines. The IL-GFE could induce anti-proliferative eﬀects as determined by MTT assay. However, IL-GFE was found to have slight anti-pro-liferation eﬀect on the normal VERO cells at markedly higher doses of 100 μg/mL compared with cancer cells. Previous studies have reported that Graviola fruit extract induced cytotoxicity toward human prostate cancer PC-3 cells, pancreatic tumor PACA-2, lung carcinoma A-549 and human hepatoma HepG2 cell lines with comparable results to our study (Consolacion et al., 2012; Sun et al., 2014, 2016, 2017). Moreover, the representative photographs of the untreated and IL-GFE treated MCF-7 cells at diﬀerent times showed a reduction in cells density. The spe-cific morphological and biochemical changes such as cell shrinkage, membrane blebbing, chromatin condensation, and DNA fragmentation are indicating the increase of apoptotic cells (Pieme et al., 2014).
To understand the anti-proliferative and cytotoxicity eﬀect of IL-GFE, the present experimental study focused on a general kinetics model that would predict the inhibitory eﬀects of IL-GFE on MCF-7 cells growth. From the growth kinetics curve, the number of generations was significantly reduced for the IL-GFE-treated MCF-7 compared with the control. A general model for breast adenocarcinoma MCF-7 cell line in response to IL-GFE treatment was developed. It is expected that an ef-fective treatment may reduce the rate of cell proliferation (cytostatic eﬀect) and raise the rate of cell death (cytotoxic eﬀect). Most che-motherapeutic agents interfere with cell division processes and disturb cell cycle regulation, either by damaging DNA or interfering with DNA synthesis (O'Reilly et al., 1997). This is proved by the doubling time when expanded from 39.09 h for the control to 86.00 h for the IL-GFE-treated MCF-7, indicating a cytotoxic eﬀect. On the other hand, Taxol had reduced the doubling time of MCF-7 cell growth to 25.62 h com-pared to the control. The exponential phase demonstrates the cytostatic eﬀects (Fig. 5A) while cytotoxic eﬀects were inferred at the death phase (Fig. 5B).