Chung Ho Ryu

Characterization of a profile of the anthocyanins isolated from Vitis coignetiae Pulliat and their anti-invasive activity on HT-29 human colon cancer cells.

We isolated anthocyanins from fruits of Vitis coignetiae Pulliat, characterized the anthocyanin profile, and investigated the anti-invasive effects of the anthocyanins on human colon cancer cells. The anthocyanins inhibited cell invasion in a dose-dependent manner, as measured by Matrigel invasion assays, by suppression of matrix metalloproteinase (MMP)-2 and MMP-9 expression. The anti-invasive activity of the anthocyanins was associated with modulation of constitutive nuclear factor kappaB (NF-kappaB) activation. The activation of NF-kappaB triggered by tumor necrosis factor-alpha was also inhibited by the anthocyanins through suppression IkappaBalpha phosphorylation. AIMs inhibited the expression of NF-kappaB-regulated proteins. In conclusion, this study suggested that the anthocyanins isolated from fruits of V. coignetiae Pulliat should have anti-invasive activities on human colon cancer cells and the activities should be related to the inhibition of NF-kappaB-regulated proteins such as MMP-2 and MMP-9 expression through the inhibition of NF-kappaB activation.

Induction of Apoptosis and Inhibition of Invasion in Human Hepatoma Cells by Anthocyanins from Meoru

Anthocyanins belong to a class of flavonoids exhibiting antioxidant and anti‐inflammatory actions as well as a variety of chemotherapeutic effects. However, little is known about the cellular and molecular mechanism of anticancer activity. In this study, we investigated if the anthocyanins (delphinidin‐3,5‐diglucoside: cyanidin‐3,5‐diglucoside: petunidin‐3,5‐diglucoside: delphinidin‐3‐glucoside: malvdin‐3,5‐diglucoside: peonidin‐3,5‐diglucoside: cyanidin‐3‐glucoside: petunidin‐3‐glucoside: peonidin‐3‐ glucoside: malvidin‐3‐ glucoside = 27: 63: 8.27: 1:2.21: 6.7: 1.25: 5.72: 1.25) isolated from meoru (Vitis coignetiae Pulliat) exerted antiproliferative and anti‐invasive and apoptotic effects on human hepatoma Hep3B cells. It was found that the anthocyanins could inhibit cell growth by 75% at the concentration of 400 μg/mL for 48 h. Flow cytometric analysis showed that the anthocyanins increased the amount of DNA fragments (sub‐G1 fraction) in a dose‐dependent manner, which is closely related to mitochondrial dysfunction and reduction in antiapoptotic proteins (Bcl‐2, xIAP, cIAP‐1, and cIAP‐2). The anthocyanins also significantly inhibited the migration and invasion of Hep3B cells through a matrigel‐coated chamber. Taken together this study indicates that the anthocyanins from meoru have antiproliferative and anti‐invasive effects and may induce apoptosis through the activation of the mitochondrial pathway and inhibition of antiapoptotic proteins. This study provides evidence that the anthocyanins isolated from meoru might be useful in the treatment of human hepatitis B‐associated hepatoma.

The flavonoid morin from Moraceae induces apoptosis by modulation of Bcl-2 family members and Fas receptor in HCT 116 cells

It is evident based on literature that flavonoids from fruit can safely modulate cancer cell biology and induce apoptosis. Therefore, we investigated the anticancer activity of morin, a flavonoid which is plentiful in twigs of mulberry focusing on apoptosis, and its mechanisms. Morin upregulated the Fas receptor, and activates caspase-8, -9 and -3 in HCT-116 cells. Morin also activates Bid, and induced the loss of mitochondrial membrane potential (MMP, ∆Ψm) with Bax protein activation and cytochrome c release. In addition, morin induced ROS generation which was not blocked by N-acetylcysteine. Morin also suppressed Bcl-2 and cIAP-1, anti-apoptotic proteins, which may contribute to augmentation of morin-triggered apoptosis. As an upstream signaling pathway, suppressed Akt activity by morin was associated to apoptosis. This study suggests that morin induces caspase-dependent apoptosis through extrinsic pathway by upregulating Fas receptor as well as through the intrinsic pathway by modulating Bcl-2 and IAP family members, and ROS generation, and that Akt is the critical upstream signaling that regulates the apoptotic effect of morin in human colon cancer HCT-116 cells.

Induction of Apoptosis by Linoleic Acid Is Associated with the Modulation of Bcl-2 Family and Fas/FasL System and Activation of Caspases in AGS Human Gastric Adenocarcinoma Cells

In this study, we investigated the effects of linoleic acid (LA), a polyunsaturated fatty acid found in most vegetable oils and certain food products, on the growth of AGS human gastric adenocarcinoma cells. LA treatment resulted in a concentration-dependent growth inhibition of AGS cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, chromatin condensation, and the accumulation cells in the sub-G1 phase. LA treatment induced cyclin-dependent kinase inhibitor p21 in a p53-independent manner; however, this compound did not affect the cell cycle distribution. Reverse transcription-polymerase chain reaction and western blot analyses showed that treating the cells with LA caused the up-regulation of pro-apoptotic Bax expression and the down-regulation of anti-apoptotic Bcl-2 expression. The apoptosis of AGS cells by LA was found to be associated with an elevated Fas and Fas ligand expression in a concentration-dependent manner. Furthermore, a proteolytic activation of caspases (3, 8, and 9), and degradation/cleavage of poly(ADP-ribose) polymerase and phospholipase C-gamma 1 protein were noted in LA-treated AGS cells. The present results indicate that the Fas/Fas ligand pathway might be involved in LA-induced apoptosis of AGS cells.

Polyphenols isolated from Allium cepa L. induces apoptosis by suppressing IAP-1 through inhibiting PI3K/Akt signaling pathways in human leukemic cells.

Allium cepa Linn is commonly used as supplementary folk remedy for cancer therapy. Evidence suggests that Allium extracts have anti-cancer properties. However, the mechanisms of the anti-cancer activity of A. cepa Linn are not fully elucidated in human cancer cells. In this study, we investigated anti-cancer effects of polyphenols extracted from lyophilized A. cepa Linn (PEAL) in human leukemia cells and their mechanisms. PEAL inhibited cancer cell growth by inducing caspase-dependent apoptosis. The apoptosis was suppressed by caspase 8 and 9 inhibitors. PEAL also up-regulated TNF-related apoptosis-inducing ligand (TRAIL) receptor DR5 and down-regulated survivin and cellular inhibitor of apoptosis 1 (cIAP-1). We confirmed these findings in other leukemic cells (THP-1, K562 cells). In addition, PEAL suppressed Akt activity and the PEAL-induced apoptosis was significantly attenuated in Akt-overexpressing U937 cells. In conclusion, our data suggested that PEAL induced caspase-dependent apoptosis in several human leukemic cells including U937 cells. The apoptosis was triggered through extrinsic pathway by up-regulating DR5 modulating as well as through intrinsic pathway by modulating IAP family members. In addition, PEAL induces caspase-dependent apoptosis at least in part through the inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. This study provides evidence that PEAL might be useful for the treatment of leukemia.

Suppressive Effect on Lipopolysaccharide-Induced Proinflammatory Mediators by Citrus aurantium L. in Macrophage RAW 264.7 Cells via NF-κB Signal Pathway

Citrus fruits have been used as an edible fruit and a traditional medicine since ancient times. In particular, the peels of immature citrus fruits are used widely in traditional herbal medicine in Korea, as they are believed to contain bioactive components exerting anti-inflammatory activity. This study examined whether the crude methanol extract of Citrus aurantium L. (CME) has a suppressive effect on inducible enzymes and proinflammatory cytokines by inhibiting the NF-κB pathway in LPS-stimulated macrophage RAW 264.7 cells. The cells were pretreated with the indicated concentrations of CME (5, 10, 20, and 50 μg/mL) and then treated with LPS (1 μg/mL). The results showed that CME (10, 20, and 50 μg/mL) inhibited the LPS- (1 μg/mL) induced mRNA and protein expression of iNOS in macrophage Raw 264.7 cells. In addition, the expression of COX-2 was inhibited at the mRNA and protein levels by CME in a dose-dependent manner. The mRNA expression of proinflammatory cytokines, such as TNF-α and IL-6, were markedly reduced by CME (10, 20, and 50 μg/mL). Moreover, CME clearly suppressed the nuclear translocation of the NF-κB p65 subunits, which was correlated with its inhibitory effect on I-κB phosphorylation. These results suggest that CME has anti-inflammatory properties by modulating the expression of COX-2, iNOS, and proinflammatory cytokines, such as TNF-α and IL-6, in macrophage RAW 264.7 cells via the NF-κB pathway.

Morin, a Flavonoid from Moraceae, Induces Apoptosis by Induction of BAD Protein in Human Leukemic Cells

Evidence suggests that phytochemicals can safely modulate cancer cell biology and induce apoptosis. Here, we investigated the anti-cancer activity of morin, a flavone originally isolated from members of the Moraceae family in human leukemic cells, focusing on apoptosis. An anti-cancer effect of morin was screened with several human leukemic cell lines. U937 cells were most sensitive to morin, where it induced caspase-dependent apoptosis in a dose-dependent manner. It also induced loss of MMP (ΔΨm) along with cytochrome c release, down-regulated Bcl-2 protein, and up-regulated BAX proteins. The apoptotic activity of morin was significantly attenuated by Bcl-2 augmentation. In conclusion, morin induced caspase-dependent apoptosis through an intrinsic pathway by upregulating BAD proteins. In addition, Bcl-2 protein expression is also important in morin-induced apoptosis of U937 cells. This study provides evidence that morin might have anticancer properties in human leukemic cells.

Anti-invasive activity of anthocyanins isolated from Vitis coignetiae in human hepatocarcinoma cells.

We investigated anti-invasive effects of the anthocyanins from fruits of Vitis coignetiae Pulliat (known as meoru in Korea) on human hepatoma Hep3B cells. The anthocyanins inhibited cell invasion in a dose-dependent manner as measured by Matrigel (BD Biosciences, San Jose, CA, USA) invasion assays. They also inhibited expression of matrix metalloproteinase (MMP)-2 and MMP-9 and activation of nuclear factor kappaB (NF-kappaB) stimulated by tumor necrosis factor alpha. Taken together, the results of this study indicate that the anthocyanins isolated from fruits of V. coignetiae Pulliat have anti-invasive effects on human hepatoma Hep3B cells and inhibit MMP-2 and MMP-9 gene expression at least in part through the inhibition of NF-kappaB activation.

Pachymic Acid Induces Apoptosis of EJ Bladder Cancer Cells by DR5 Up-Regulation, ROS Generation, Modulation of Bcl-2 and IAP Family Members

Pachymic acid (PA) is a lanostane‐type triterpenoid derived from Poria cocos mushroom that possess various biological effects such as anti‐cancer, antiinflammatory and anti‐metastasis effects. In this study, we investigated the anti‐cancer effects of PA in EJ bladder cancer cells. The results showed that PA significantly inhibited proliferation of EJ cells in a dose‐dependent manner. PA induced accumulation of sub‐G1 DNA content (apoptotic cell population), apoptotic bodies and chromatin condensation and DNA fragmentation in EJ cells in a dose‐dependent manner. PA also induces activation of caspase‐3, ‐8 and ‐9, and subsequent cleavage of poly (ADP‐ribose) polymerase, and significantly suppressed the inhibitor of apoptosis protein family proteins in a dose‐dependent manner. Furthermore, PA activates Bid and induced the loss of mitochondrial membrane potential (ΔΨm) with up‐regulated pro‐apoptotic proteins (Bax and Bad), down‐regulated anti‐apoptotic proteins (Bcl‐2 and Bcl‐xL) and cytochrome c release. In turn, PA increased the generation of reactive oxygen species (ROS); also, the ROS production was blocked by N‐acetyl‐L‐cysteine. The expressions of TNF‐related apoptosis inducing ligand and death receptor 5 were up‐regulated by PA in a dose‐dependent manner, suggesting extrinsic pathway also involved in PA‐induced apoptosis. This study provides evidence that PA might be useful in the treatment of human bladder cancer. Copyright

Piceatannol inhibits mast cell-mediated allergic inflammation

Piceatannol is a phenolic stilbenoid and a metabolite of resveratrol which is found in red wine. Piceatannol (PIC) commonly exhibits anti-inflammatory, antiplatelet and antiproliferative activity. In the present study, the anti-allergic and anti-inflammatory mechanisms of PIC were investigated by examining the effects of PIC on pro‑inflammatory cytokine release and phosphorylation of mitogen-activated protein (MAP) kinases (ERK, JNK and p38) in a human mast cell line. PIC dose-dependently inhibited compound 48/80-induced systemic anaphylaxis and immunoglobulin E-mediated local allergic reactions. PIC reduced the immunoglobulin E (IgE)-mediated local allergic reaction and attenuated histamine release from rat peritoneal mast cells. Histamine and β-hexosaminidase release was markedly decreased dose-dependently by PIC treatment in RBL-2H3 cells. PIC treatments of HMC-1 cells definitely reduced mRNA expression and the release of the pro‑inflammatory cytokines, tumor necrosis factor-α and interleukin-8. MAP kinase phosphorylation was also strongly decreased dose-dependently following PIC treatment. PIC regulated the production of cytokines and histamine in phorbol 12-myristate 13-acetate plus A23187-stimulated mast cells. Thus, PIC may alleviate allergic inflammation and may be a useful therapeutic agent for allergic diseases.