# The Suppressive Activity of Water Minosa Extract on Human Gastric Cancer Cells
## 4 Discussion  
Cancer cells were charactered due to the inactivated apoptosis, the uncontrolled growth, and metastasis [14]. Therefore, suppression of growth, inhibition of metastasis, and induction of apoptosis in cancer cells were considered as potential strategies for the prevention and treatment of cancers [15]. In this sense, various herbs have been screened to be effective in the inhibition of the proliferation of cancer cells [15; 16]. In this study, the MTT assay revealed that the EtOAc extract containing high total phenolic content exhibited potential inhibition of the proliferation of human gastric cancer cells. Numerous studies have suggested that polyphenols such as flavonoids, phenolic acids, lignans, and stillenes have the capacity to inhibit tumor generation and thus could be used for the control of cancer [17; 18]. Therefore, the potential inhibitory activity of EtOAc extract on cell proliferation may be due to its phenolic content. In particular, the inhibitory effect of the EtOAc extract of _N. oleracea_ on the proliferation of gastric cancer cells was observed to be higher than that of the aqueous extracts of zizizphora, ginger, saffron, aloe vera [19], dandelion (_Taraxacum_ spp.) [20], and _Curcuma manga_[21]. Hence, _N. oleracea_ was also considered as a potential agent for targeting gastric cancer cell growth. In addition, anticancer therapy not only targets cell proliferation but also focuses on suppression of cell migration and invasion [22]. Notably, MMP-2 and MMP-9 were evidenced to play a crucial role in degrading extracellular matrices, thus facilitating the invasion and metastasis of cancer cells [23]. The inhibition of MMP-2 and MMP-9 may contribute to the suppression of cancer cell migration, bringing about the prevention of metastasis [24; 25]. In this study, the EtOAc extract of _N. oleracea_ was shown to inhibit MMP-2 and MMP-9 and suppress the cell migration at a low concentration of treatment (50 \(\mathrm{\SIUnitSymbolMicro g}\)/mL). As a result, _N. oleracea_ has potential in suppression of the proliferation at the primary sites as well as the prevention of migration of gastric cancer cells to new sites. Figure 4: EtOAc extract induced apoptosis in BGC-823 cells. The cells were exposed by EtOAc extract, and total RNA was then collected. The expression levels of caspase-3, -8, -9, and Bax were identified using real-time PCR (**A**) and gel electrophoresis (**B**). GAPDH was used as an internal control. It was observed that the EIOAc extract of _N. olercea_ exhibited a cytotoxic effect toward gastric cancer cells, thus inhibiting cell growth and migration. However, this result should clearly indicate whether its cytotoxic effect was due to necrosis or apoptosis. Certainly, a natural product is considered as a potential anti-cancer agent when its cytotoxic effect is due to the promotion of apoptosis in the cancer cells [26; 27]. Apoptosis is a caspase-mediated programmed cell death, leading to chromosome condensation, nuclear fragmentation, and membrane blebbing [28]. First, the binding of the Fas ligand, TNF-\(\alpha\), or TRAIL to the corresponding death receptors triggers the activation of caspase-8, which then activates caspase-3. Moreover, the caspase-8 activation also results in conformational changes in Bax, which causes the release of cytochrome C from mitochondria. Subsequently, the combination of cytochrome C and Apaf-1 activated caspase-9 and caspase-3 [28]. Finally, the activation of caspase-3 induces the process of apoptosis [29]. As a result, the induction of apoptosis due to the upregulation of caspase-3, -8, -9, and Bax activation in cancer cells can trigger the process of cell death. In this study, the treatment of the EIOAc extract of _N. olercea_ caused a significant increase in the gene expression levels of caspase-3, -8, -9, and Bax in gastric cancer cells.