The ethyl acetate extract of Phellinus linteus grown on germinated brown rice induces G0/G1 cell cycle arrest and apoptosis in human colon carcinoma HT29 cells

Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, kwangjin-gu, Seoul, 143-701, Korea.
Phytotherapy Research (Impact Factor: 2.66). 01/2009; 24(7):1019-26. DOI: 10.1002/ptr.3064
Source: PubMed


It is well known that Phellinus linteus has a variety of biological functions, such as antitumor and immunomodulating activities. In our previous studies, we developed a P. linteus grown on germinated brown rice (PBR) and found that organic solvent extracts of PBR possessed immunomodulating activity to regulate a balance of cytokine network in mice. The components of PBR are ergosterol peroxide, gamma-aminobutyric acid (GABA) and Beta-glucan. In this study, we demonstrate that an organic solvent extract of P. linteus grown on PBR induced apoptotic cell death through the induction of G(0)/G(1) arrest of cell cycle and the apoptosis via DNA fragmentation in human colon carcinoma HT-29 cells. Cell death induced by the extract of P. linteus grown on PBR was shown to be associated with the upregulation of p21(CIP1/WAF1), the downregulation of cyclin D1, anti-apoptotic protein, Bcl-2, the release of cytochrome c, and the activation of caspase-9, caspase-3 and caspase-8. This study suggests that the ethyl acetate extract of P. linteus grown on PBR induces apoptosis accompanied by cell cycle arrest at G(0)/G(1) phase and regulates apoptosis-regulatory proteins, which may be applicable to anticancer therapy.

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    • "Although most studies have focused on polysaccharides as active ingredients responsible for the antitumor effects of P. linteus [10] [11] [12] , our previous studies, interestingly, indicated that the anticancer activity of PB was observed in ethyl acetate extract, but not in boiled-water [8] . These observations suggested that the major anticancer constituent of PB might be retained in the nonpolar organic solvent fraction rather than in the polysaccharide-rich aqueous fraction. "
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    ABSTRACT: To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate (EtOAc) extract of Phellinus linteus grown on germinated brown rice (PB). EtOAc extract of PB was partitioned with n-hexane, EtOAc, and water-saturated n-butanol. Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR, respectively. Cytotoxicity against HT-29 cells was tested by SRB assay. The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line. Atractylenolide I, a eudesmane-type sesquiterpene lactone, a major anticancer substance of PB, was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC. This structure was elucidated by one- and two-dimensional NMR spectroscopic data. Atractylenolide I has not been reported in mushrooms or rice as of yet. The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells. Atractylenolide I might contribute to the anticancer effect of PB.
    Full-text · Article · Oct 2013 · Asian Pacific Journal of Tropical Biomedicine
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    • "Lysates were centrifuged at 14,000 × g for 10 min at 4 • C. The protein concentrations were determined using a BCA Protein Assay kit (Bio-Rad Laboratories, Hercules, CA) as previously described (Park et al., 2010b). Then, 20 ␮g aliquots of protein were subjected to electrophoresis on 4–15% gradient polyacrylamide gels and electrophoretically transferred onto polyvinylidene fluoride Fig. 1. "
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    ABSTRACT: Cordyceps militaris has been used as a traditional herbal medicine for treating allergy in East Asia. We investigated the anti-allergic efficacy of Cordyceps militaris and its mechanism of action. β-Hexosaminidase release of mast cells, a key parameter of degranulation, was evaluated. Anti-allergic potential of Cordyceps militaris was studied using passive cutaneous anaphylaxis (PCA) in vivo. The anti-allergic mechanism of Cordyceps militaris was investigated by immunoblotting analysis, RT-PCR and other biological approaches in mast cells. GSCM EtOAc extract (GSCME) inhibited antigen-induced degranulation with a IC50 value (28.5 μg/ml) in RBL-2H3 cells and antigen-induced passive cutaneous anaphylaxis (PCA) response with a ED50 value (665 mg/kg) in vivo. The release of interleukin (IL-4) and tumor necrosis factor (TNF-α were decreased by GSCME in RBL-2H3 cells. In order to elucidate the anti-allergic mechanisms of GSCME in mast cells, we examined the activated levels of signaling molecules. GSCME inhibited the phosphorylation Syk, ERK, p38 and JNK expression. Identified genistein, daidzein, genistein 7-O-β-d-glucoside 4″-O-methylate, genistein 4'-O-β-d-glucoside 4″-O-methylate, glycitein 7-O-β-d-glucoside 4″-O-methylate, daidzein 7-O-β-d-glucoside 4″-O-methylate and adenosine in GSCME, inhibited antigen-induced degranulation in RBL-2H3 cells. Our study suggests that GSCME might be used as a therapeutic agent for allergic diseases.
    Preview · Article · Mar 2011 · Journal of ethnopharmacology
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    ABSTRACT: Phellinus linteus and Panax ginseng have been widely used as traditional herbal medicines to treat various diseases including cancer in East Asia. The present study sought to investigate the possible mechanism in anti-proliferative effect of Phellinus linteus that was grown on Panax ginseng (PGP) on B16F10 melanoma cells. The anti-proliferative effect of PGP on B16F10 was evaluated by CCK-8 assays. Apoptotic cells were detected by flow cytometry analysis. The proteins involved in apoptosis and cellular differentiation were assessed by immunoblot analysis. Ginsenosides contents of PG or PGP were analyzed using HPLC. The ethyl acetate fraction (EtOAc) of PGP exhibited the strongest anti-proliferative activity among PGP fractions (butanol or water) on B16F10 cells. PGP EtOAc extract showed stronger inhibitory effect than Panax ginseng (PG) or Phellinus linteus (PL) EtOAc extract on B16F10 melanoma cell proliferation. PGP EtOAc extract induced the dendrite-like structures and the melanin production in B16F10 cells. PGP EtOAc extract increased a sub-G1 cell population through inducing p53/p21 and activated caspase-8 protein expression in B16F10 cells. Notably, PGP EtOAc extract contained ginsenosides Rd, Rg3, Rb2, Rg1 and Rb1 more than PG EtOAc extract. Rd and Rg3 significantly inhibited B16F10 cell proliferation. Our data suggest that PGP EtOAc extract inhibits B16F10 cell proliferation through inducing apoptosis and cellular differentiation.
    Preview · Article · Oct 2010 · Journal of ethnopharmacology
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