Hwa-Seung Yoo

Induction of apoptosis and inhibition of telomerase activity in human lung carcinoma cells by the water extract of Cordyceps militaris

Cordyceps militaris is well known as a traditional medicinal mushroom and is a potentially interesting candidate for use in cancer treatment. In this study, the potential of the water extract of C. militaris (WECM) to induce apoptosis in human lung carcinoma A549 cells and its effects on telomerase activity were investigated. The growth inhibition and apoptosis induction by WECM treatment in A549 cells was associated with the induction of Fas, catalytic activation of caspase-8, and Bid cleavage. Activation of caspases, downregulation of anti-apoptotic Bcl-2 expression, and upregulation of pro-apoptotic Bax protein were also observed in WECM-treated cells. However, the cytotoxic effects and apoptotic characteristics induced by WECM were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role that caspase-3 plays in the process. In addition, WECM exerted a dose-dependent inhibition of telomerase activity via downregulation of human telomerase reverse transcriptase (hTERT), c-myc and Sp1 expression. Taken together, the data from this study indicate that WECM induces the apoptosis of A549 cells through a signaling cascade of death receptor-mediated extrinsic and mitochondria-mediated intrinsic caspase pathways. It was also conclude that apoptotic events due to WECM were mediated with diminished telomerase activity through the inhibition of hTERT transcriptional activity.

Anti-inflammatory effects of Polygala tenuifolia root through inhibition of NF-κB activation in lipopolysaccharide-induced BV2 microglial cells.

ETHNOPHARMACOLOGICAL RELEVANCE The root of Polygala tenuifolia Willd is a well-known traditional Oriental medicine and has been prescribed for treatment of dysfunction in memorial systems and various brain inflammatory diseases. The present study was designed to validate the anti-inflammatory effects of the water extract of Polygala tenuifolia root (WEPT). MATERIALS AND METHODS The anti-inflammatory properties of WEPT were studied using lipopolysaccharide (LPS)-stimulated murine BV2 microglia model. As inflammatory parameters, the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin E(2) (PGE(2)), tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were evaluated. We also examined the extracts effect on the activity of nuclear factor-kappaB (NF-κB), and toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (Myd-88) expression. RESULTS WEPT suppressed LPS-induced production of NO, PGE(2), and expression of iNOS and COX-2 in a dose-dependent manner, without causing cytotoxicity. It also significantly reduced generation of proinflammatory cytokines, including IL-1β and TNF-α. In addition, WEPT suppressed NF-κB translocation by blockade of IkappaB-α (IκB-α) degradation and inhibited TLR4 and Myd-88 expression in LPS-stimulated BV2 cells. CONCLUSIONS These results indicate that the inhibitory effects of WEPT on LPS-stimulated inflammatory mediator production in BV2 microglia are associated with suppression of the NF-κB and toll-like receptor signaling pathways. Therefore, Polygala tenuifolia extracts may be useful in treatment of neurodegenerative diseases by inhibition of inflammatory mediator production in activated microglia.

Mountain ginseng extract exhibits anti-lung cancer activity by inhibiting the nuclear translocation of NF-κB

Administration of mountain ginseng (MG) extract can restore advanced cancer to a normal state. To elucidate the mechanism by which MG extract prevents the progression of lung cancer, the processes of proliferation and death of lung cancer cells (A549) were examined after treatment with MG extract. Butanol-extracted MG (BX-MG) showed a high inhibitory effect (IC(50) = 2 mg/ml) by attenuating proliferation and inducing apoptosis in lung cancer cells. By HPLC-UV analysis of BX-MG, ginsenosides, Rb1 was identified as the most abundant ginsenoside, followed by Rg1, Re, Rc and Rb2. BX-MG induced caspase-3 dependent apoptosis by inhibiting NF-κB. In addition, BX-MG activated p53 and p21, resulting in the attenuated proliferation of A549 cells. Reduced activity of the NF-κB promoter and increased activity of the p53 promoter indicate that BX-MG regulates apoptosis at the level of transcription in lung cancer cells. Furthermore, BX-MG blocked the nuclear translocation of RelA and the associated reduction in surviving. These results suggest that BX-MG inhibits lung cancer cell growth by activating tumor suppressors and inhibiting nuclear translocation of NF-κB.

Soluble components of hericium erinaceum induce NK cell activation via production of interleukin-12 in mice splenocytes

AbstractAim:To investigate the immunoregulatory functions of water extracts of Hericium erinaceum (WEHE) focusing on natural killer (NK) cell-based anticancer activities.Methods:Mouse splenocytes or purely isolated NK cells were stimulated with 1-100 mg/L WEHE for 24 h followed by co-culture with 51Cr-labled Yac-1 cells for 4 h, then NK cell-derived cytolytic activity was measured using a radio-release assay. Neutralizing antibodies against mouse interleukin-12 (IL-12) were added into the WEHE-stimulated splenocytes, thereafter, cytotoxicity was measured to examine the involvement of IL-12. RT-PCR and ELISA analyses were performed to confirm the induction of transcription and the translation of IL-12 and interferon-gamma (IFN-gamma) in the WEHE-treated splenocytes.Results:WEHE enhanced the cytolytic activity of total splenocytes towards Yac-1 cells in a dose-dependent manner. However, this activation was not observed when the NK cells isolated from the splenocytes were treated with WEHE. Furthermore, the treatment with antibodies against IL-12 abolished the effect of WEHE on splenocyte-derived cytolytic activity. RT-PCR and ELISA analyses showed the induction of IL-12 and IFN-gamma in the WEHE-treated splenocytes.Conclusion:WEHE indirectly activates the cytolytic ability of NK cells via the induction of IL-12 in total splenocytes, and possibly via other immuno-mediators or cellular components.

Antitumor Activity of Water Extracts From Cordyceps Militaris in NCI-H460 Cell Xenografted Nude Mice

This experimental study investigated the antitumor effect of Cordyceps militaris in NCI-H406 cell transplanted nude mice. After feeding an aqueous solution of C. militaris extracts in NCI-H460 cell xenografted nude mice for 4 weeks, we measured the size of a tumor mass and calculated the inhibition rate. We also estimated survival time and calculated mean survival time and percent increase in lifespan. Results showed that the inhibition rate of water extract of the 150 mg/kg/day C. militaris-administered group was 94.73-75.08% and that of the 300 mg/kg/day C. militaris-administered group was 85.81-73.81%. The tumor weights and volumes decreased in a dose-dependent manner. Mean survival time of the 150 mg/kg/day C. militaris-administered group was extended to 19.43 +/- 2.44 days and 5.42% increased in lifespan (ILS) and that of the 300 mg/kg/day C. militaris-administered group was 21.86 +/- 3.53 days and 18.61% ILS. The relative liver weight was significantly increased in 300 mg/kg/day C. militaris-administered group, but there was no histopathological difference. In conclusion, C. militaris, shrunk tumors and increased mouse lifespan, suggesting that C. militaris was effective in treating tumors in nude mice.

Antimetastatic and Immunomodulating Effect of Water Extracts From Various Mushrooms

This experiment was conducted to evaluate inhibitory effects against lung metastasis and promotion of splenocytes by water extracts from various mushrooms including Armillaria mellea, Grifola frondosa, Garnoderma frondosa, Codyceps militaris, Hericium erinaceus, Coriolus versicolor, Agaricus Blazei with Lycium Chinense Miller (known as M8). Analysis of carbohydrate using HPTLC showed that beta-glucan and pachyman were some of the major components of M8. Oral administration of M8 resulted in a dose-dependent tendency to inhibit lung metastasis after intravenous injection of colon26-L5 cells. Treatment with M8 resulted in a significant increase of T cell and B cell mitogenic stimuli. The population of CD3, CD19, CD4, and CD8 positive cells increased in a dose dependent manner of M8 administration. However, no significant results were obtained from the population of Mac-1 and NK1.1 positive cells. Oral administration of M8 resulted in the increased production of IFN-gamma and IL-4 by splenocytes stimulated with Con A compared with untreated controls. These results show that M8 has antitumor activities which may be useful as an antimetastatic agent.

Aqueous extract of Tribulus terrestris Linn induces cell growth arrest and apoptosis by down-regulating NF-κB signaling in liver cancer cells

ETHNOPHARMACOLOGICAL RELEVANCE A medicinal herb Tribulus terrestris Linn has been used to treat various diseases including hepatocellular carcinoma. The aim of the present study was to investigate the anticancer activity of Tribulus terrestris Linn (TT) in liver cancer cells. MATERIALS AND METHODS The antitumor activity of aqueous TT extract was analyzed by testing the cytotoxicity and the effect on clonogenecity in HepG2 cells. Apoptosis and cell cycle arrest induced by TT were dissected by flow cytometry and its inhibitory effect on NF-κB activity was determined by analyzing the expression levels of NF-κB/IκB subunit proteins. The suppression of NF-κB-regulated gene expression by TT was assessed by RT-PCR. RESULTS TT extract repressed clonogenecity and proliferation, induced apoptosis, and enhanced accumulation in the G0/G1 phase of liver cancer cells. It also turned out that TT extract inhibited NF-κB-dependent reporter gene expression and NF-κB subunit p50 expression, while it enhanced the cellular level of IκBα by inhibiting the phosphorylation and degradation of IκBα. In addition, IKK activity was inhibited in a dose-dependent manner. Furthermore, TT extract suppressed the transcription of genes associated with cell cycle regulation, anti-apoptosis, and invasion. CONCLUSION These data showed that TT extract blocks proliferation and induces apoptosis in human liver cancer cells through the inhibition of NF-κB signaling. Aqueous TT extract can be used as an anticancer drug for hepatocellular carcinoma patients.

DMNQ S-64 Induces Apoptosis via Caspase Activation and Cyclooxygenase-2 Inhibition in Human Nonsmall Lung Cancer Cells

Abstract:  Shikonin has been reported to induce apoptosis and inhibit angiogenesis in vivo and in vitro. 6‐(1‐propoxyiminoalkyl)‐5,8‐dimethoxyoxy 1,4‐naphtoquinone S‐64 (DMNQ S‐64) was synthesized as a shikonin derivative. In this article, the underlying apoptotic mechanism of DMNQ S‐64 was examined. DMNQ S‐64 exerted cytotoxicity against A549 lung carcinoma cells with IC50 of 27.3 μM. Apoptotic bodies were observed in DMNQ S‐64‐treated A549 cells by 4′‐6‐diamidino‐2‐phenylindole (DAPI) staining assay. DMNQ S‐64 also increased sub‐G1 DNA portion in a concentration‐dependent manner by flow cytometric analysis. Western blotting has revealed that DMNQ S‐64 effectively activates the expression of caspase 8, 9, and 3, cleaves poly (ADP‐ribose) polymerase, and increases the ratio of Bax/Bcl‐2. Furthermore, cytochrome c was released in a concentration‐dependent manner by DMNQ S‐64. Similarly, DMNQ S‐64 significantly increased caspase 3 activity by enzyme‐linked immunosorbent assay (ELISA). It also significantly inhibited the level of prostaglandin E2 (PGE2) by ELISA and downregulated the expression of cyclooxygenase‐2 (COX‐2) in a concentration‐dependent manner. Taken together, DMNQ S‐64 may exhibit cytotoxicity against A549 cells via caspase activation and COX‐2 inhibition.

Effects of Sweet Bee Venom Pharmacopuncture Treatment for Chemotherapy-Induced Peripheral Neuropathy A Case Series

Objective. This is a case series reporting safety and degree of response to 1 dose level of sweet bee venom pharmacopuncture (SBVP) or melittin as a symptom-control therapy for chemotherapy-induced peripheral neuropathy (CIPN). Setting. All treatments were conducted at the East West Cancer Center (EWCC), Dunsan Oriental Hospital, Daejeon University, Republic of Korea, an institution that uses complementary therapies for cancer patients. Methods. Five consecutive patients with CIPN were referred to the EWCC from March 20, 2010, to April 10, 2010. Patients with World Health Organization Chemotherapy-Induced Peripheral Neuropathy (WHO CIPN) grade 2 or more were treated with SBVP for 3 treatment sessions over a 1-week period. Measures of efficacy and safety. Validated Visual Analog System (VAS) pain scale, WHO CIPN grade, and Functional Assessment of Cancer Therapy–General (FACT-G) were compared before and after the 1-week course of treatment. To ensure the safety of SBVP, pretreatment skin response tests were given to patients to avoid any potential anaphylactic adverse effects. All patients were closely examined for any allergenic responses following each treatment session. Results. One patient discontinued treatment after the first session, and 4 patients completed all treatment sessions. Using each patient as their own comparator, marked improvements of VAS, WHO CIPN grade, and physical section scores of FACT-G were seen in 3 patients. Most important, there were no related adverse side effects found. Conclusion. This safety results of the SBVP therapy merits further investigations in a larger size trial for it to develop into a potential intervention for managing CIPN symptoms. This study will be extended to a dose–response evaluation to further establish safety and response, prior to a randomized trial.

Lactobacillus casei Extract Induces Apoptosis in Gastric Cancer by Inhibiting NF-κB and mTOR-Mediated Signaling

Lactobacillus casei extract (LBX) has been reported to prevent gastric cancer, but the underlying mechanism remains unclear. The proliferation and cell death of gastric cancer KATO3 cells were examined after treatment with LBX for various times and at various doses. LBX inhibited the growth of gastric cancer cells and induced apoptosis by inactivating NF-κB promoter activity. Apoptosis induced by LBX, however, is not directly associated with the intrinsic mitochondrial pathway. Immunoblot analysis revealed that LBX decreased the expressions of NF-κB and IκB. The reduced NF-κB levels led to the decreased phosphorylation of mTOR signaling components, such as PI3K, Akt, and p70S6 kinase. These results showed for the first time that LBX induced apoptosis in gastric cancer cells by inhibiting NF-κB and mTOR-mediated signaling.