After 24?h of incubation, extract (250?g/mL) was added and the plates were incubated at 37C for 48?h

After 24?h of incubation, extract (250?g/mL) was added and the plates were incubated at 37C for 48?h. induction of apoptosis involved the intrinsic pathways of the cells. Take the results, we suggest that extract has anti-gastric cancer activity and may be a potential therapeutic candidate for gastric cancer. is Etretinate a medicinal herb that has been traditionally used as a remedy for intestinal disorders in East Asia. has also been reported to Etretinate have various biological activities (Liu et al. 2006; Sung et al. 2011). There are two ways to use medicinally. The dried aerial parts can be used to make a tea, or the dried plant can be boiled in water (Hiramatsu et al. 2004). The tea and boiled dried plant preparations are used to treat constipation and diarrhea, respectively, and also to prevent gastritis (Liu et al. 2006). The ability of to suppress cancer cell growth is primarily mediated through the induction of apoptosis in lung adenocarcinoma (Li et al. 2013). As such, is generally used as a therapeutic agent for digestive system diseases and has an anti-cancer mechanism, but interestingly, there is no Etretinate research on its relationship with gastric cancer and the mechanism its effect on gastric cancer. Therefore, we focused on role of in gastric cancer. The failure to control cancer cell death associated with the induction of apoptosis and cell cycle arrest Etretinate is considered the main limitation of cancer therapy (Evan and Vousden 2001; Nawab et al. 2012; Ehrhardt et al. 2013; Jung et al. 2018). Apoptosis is a type of Mouse monoclonal to KID programed cell death and is a physiological homeostatic mechanism (Konopleva et al. 1999; Green 2017). As a result of apoptosis, unwanted cells are eliminated in a well-organized sequential process (Konopleva et al. 1999; Green 2017). Caspases are central components of the apoptotic machinery in the proteolytic system (Konopleva et al. 1999). Apoptosis induces the activation of caspase-3 that subsequently cleaves its substrates, including poly-(ADP-ribose) polymerase (PARP), ultimately leading to apoptosis (Los et al. 2002). The cell cycle progresses in several stagesthe G1, S, G2, and M phasesand is regulated by the activation of complexes involving cell cycle proteins (cyclins) and cyclin-dependent kinases (CDKs) (Nakanishi 2001 Barnum and OConnell 2014). Since uncontrolled CDKs are often the cause of cancer, their function is tightly regulated by cell cycle inhibitors, such as p21CIP/WAF and p27KIP1 proteins (Barnum and OConnell 2014). Therefore, cell cycle arrest can be triggered by various stimulating factors, and may result in the blockage of cell division, cell death, and/or apoptosis In this study, we confirmed the effect of on anti-cancer activity using gastric cancer cell lines. We also investigated the molecular mechanism that underlies extract-induced apoptosis and G1 cell cycle arrest against YCC-2 and SNU668 gastric cancer cells. The results indicate the value of extract for the prevention of gastric cancer Etretinate cell growth. Materials and methods Preparation of G. thunbergii methanol extract Dried was purchased from Cheongmyeong Yakcho Yeoju (Korea). It was extracted with 80% (v/v) methanol at 69C for 3?h. This crude extract was dissolved in dimethyl sulfoxide. Cell culture Six human gastric cancer cell lines (AGS, MKN-28, YCC-2, SNU-216, SNU-601, and SNU-668) were obtained from the Korea Cell Line Bank. All cells were cultured in RPMI-1640 medium (Welgene, Korea) containing 5% fetal bovine serum (Corning Costar, USA) and 1% antibiotic-antimycotic (Gibco, USA) in a 37C incubator in an atmosphere of 5% CO2. Cell proliferation assay Cell proliferation after treatment with extreact was determined using the WST-1 assay. Six human gastric cancer cells were seeded in wells of 96-well plates (1??104?cells/well). After 24?h of incubation, cells were treated with extract (0, 50, 100, 200, 300, 400, and 500?g/mL) for 24, 48, and 72?h. WST-1 solution (EZ-cytox; Daeil, Korea) was added to each well.