Effects of the Ethylacetate Extract of <i>Orostachys japonicus</i> on Induction of Apoptosis through the p53-Mediated Signaling Pathway in Human Gastric Cancer Cells

Abstract

Cancer rates are increasing dramatically, and there is currently a strong emphasis on identifying biologically active substances with anti-cancer activity from traditional herbs, as these are thought to have less adverse side-effects than conventional chemotherapy. Here, we examined the effects of extracts of Orostachys japonicus A. BERGER (O. japonicus) on cancer cell proliferation and apoptosis, and investigated the underlying signaling pathways. Dried powdered O. japonicus was extracted with 95% ethyl alcohol and fractionated with a series of organic solvents, including n-hexane (hexane), dichloromethane (DCM), ethylacetate (EtOAc), n-butanol (BuOH), and water (H 2 O). These extracts were tested for anti-cancer activity on a range of cancer cells; of all these, the EtOAc soluble fraction showed the highest anti-cancer activity, which was most marked in AGS human gastric cancer cells. The EtOAc fraction inhibited the proliferation of AGS cells in a dose-dependent and time-dependent manner, by inducing apoptosis and cell cycle arrest, as evidenced by 4,6-diamidino-2-phenylindole (DAPI) staining, annexin V-fluorescein isothiocyanate staining, propidium iodide-labeling, and DNA fragmentation assays. Western blot analysis revealed that p53 and cleaved caspase-3 proteins were up-regulated, and B cell lymphoma-2 (bcl-2) protein and pro-caspase-3 were down-regulated, but bcl-2 associated x protein (bax) protein was not regulated, in response to treatment of AGS cells with the EtOAc fraction. However, the changes of pro-caspase-3 and cleaved caspase-3 could be abolished by the pancaspase inhibitor Z-VAD-FMK. These results suggest the EtOAc fraction from O. japonicus has substantial anti-cancer activity in human gastric cancer cells.

Orostachys japonicus induce p53‐dependent cell cycle arrest through the MAPK signaling pathway in OVCAR‐3 human ovarian cancer cells

“…This fraction was analyzed using a GC‐MS system following the measurement methods detailed in our study. As a result, the following three peaks were identified: “gallic acid (4.24%), kaempferol (6.81%), and quercetin (5.08%)” (Lee et al., 2014; Ryu et al., 2012). Various studies have revealed that kaempferol and quercetin lead to apoptosis using the natural products in various human cancer cells (Lee, Szcsepanski, & Lee, 2008; Yoshida et al., 2008).…”

“…Component analysis of the EtOAc fraction (OJE) has previously been described by our team (Lee et al., 2014; Ryu et al., 2012). …”

“…Orostachys japonicus (Crassulaceae), a perennial herb employed as a medicinal plant, has been used as a folk treatment for inflammatory, febrile, hemostatic, antidote, and variable cancers (Park, Han, Park, Choi, & Choi, 2005; Ryu, Lee, Lee, & Lee, 2012). Orostachys japonicus is reported to contain 16 types of flavonoid contents.…”

“…We determined that its ability to suppress cancer cell proliferation is mediated through an apoptotic mechanism. Among the O. japonicus extracts, EtOAc fraction showed the highest anticancer activity (Lee et al., 2014; Ryu et al., 2012). To our knowledge, there are no reports on anti‐cancer activity of the ethyl acetate fraction from O. japonicus extract (OJE) in human ovarian cancer cell lines.…”

Aqueous extract of Orostachys japonicus A. Berger exerts immunostimulatory activity in RAW 264.7 macrophages

“…Orostachys japonicus A. Berger (Crassulaceae) (OJ), also known as Wa-song in Korea, is used as a folk medication; its main habitat is on the surface of mountain rocks in Korea and Japan. OJ is a medicinal plant with immunoregulatory, anti-febrile, antidote, and anticancer activities (Jin Kwon and Kwangsoo Han, 2004;Kim et al, 2011;Lee et al, 2011;Ryu et al, 2014;Ryu et al, 2012;Shin et al, 2013;Yoon et al, 2000). It is reported that OJ increased CD4-and CD8-T cells and the production of interferon γ in BALB/c mice (Jin Kwon and Kwangsoo Han, 2004).…”