Yeon-Weol Lee

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.

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.

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.

Identification of Target Genes Involved in the Antiproliferative Effect of Enzyme-Modified Ginseng Extract in HepG2 Hepatocarcinoma Cell.

Ginsenosides are ginseng saponins, which are the major biologically active components of Panax ginseng, often metabolized by intestinal bacteria into more effective forms. In this study, we found that the antiproliferative activity of ginseng increased after enzymatic processing of ginseng saponin (50% inhibitory concentration [IC50], >30 μg/mL), which may be the result of the accumulation of minor saponins, such as Rh1, Rg3, compound K, and PPT constituents in ginseng saponin. Using the Agilent PrimeView Human Gene Expression Array, we found that the expression of several genes involved in apoptosis (caspase-4, Annexin A2, HSPA9, AIFM1, UQCRC2, and caspase-7) were increased in HepG2 human hepatocarcinoma cells after their treatment with enzyme-modified ginseng extract (EMGE). Furthermore, several genes implicated in cell cycle progression (CDCA3, CDCA8, CABLES2, CDC25B, CNNM3, and CCNK) showed decreased expression in HepG2 cells treated with EMGE. Finally, from flow cytometric analysis, we found that EMGE-treated HepG2 cells showed increased apoptotic sub-G1 population (24%), compared with that observed in DMSO-treated control cells (1.6%). Taken together, our results suggest that EMGE induces anticancer activity through the induction of apoptosis-related genes and cell cycle arrest via decreased expression of cell cycle regulatory genes.

Anti-angiogenic effects of the water extract of HangAmDan (WEHAD), a Korean traditional medicine.

We investigated the anti-angiogenic effects of the water extract of HangAmDan (WEHAD), which is a crude extract of nine Korean medicinal substances of animal and plant origin. In human umbilical vein endothelial cells, WEHAD significantly inhibited bFGF-induced proliferation, adhesion, migration, and capillary tube formation. We used an antibody array to perform an analysis of signaling proteins, which showed up-regulated expression of various proteins including RAD51, RAD52, and p73, and down-regulated expression of pFAK. Blood vessel formation in a chick chorioallantoic membrane (CAM) treated with WEHAD was markedly reduced in length compared with a PBS-treated control group. These results suggest that inhibition of angiogenesis by WEHAD may be the mechanism of action for the anti-cancer effects of HAD.

Inhibition of cell motility and invasion by HangAmDan-B in NCI-H460 human non-small cell lung cancer cells.

Correlation between inhibition of cell motility and anti-invasive activity by the water extract of HangAmDan-B (HAD-B), a crude extract of eight Korean medicinal animals and plants, in NCI-H460 human non-small cell lung cancer (NSCLC) cells was investigated. Within the concentrations that were not cytotoxic, HAD-B induced significant concentration-dependent inhibition of cell motility and invasiveness of NCI-H460 cells. Treatment with HAD-B resulted in dose-dependent inhibition of the activities of matrix metallo-proteinase (MMP)-2 and MMP-9, and this was correlated with a decrease in expression of their mRNA and proteins, and upregulation of tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 expression. Anti-invasive activity of HAD-B was also found to be associated with increased tightness of the tight junction (TJ), as demonstrated by an increase in transepithelial electrical resistance. In addition, the present results indicated that treatment with HAD-B resulted in repression of the levels of claudin family members, which are major components of TJs that play a key role in control and selectivity of para-cellular transport. Although further studies are needed, findings from the present study indicate that TJs and MMPs are critical targets of HAD-B-induced anti-invasiveness in NCI-H460 NSCLC cells.

Chemopreventive Effect of Curcuma longa Linn on Liver Pathology in HBx Transgenic Mice

Unlike other forms of hepatocellular carcinoma (HCC), HCC induced by hepatitis B virus (HBV) infection shows a poor prognosis after conventional therapies. HBV induces liver cirrhosis and HCC. Many researchers have made efforts to find new substances that suppress the activity of HBV. Curcuma longa Linn (CLL) has been used for traditional medicine and food in Asia, especially in India, and has shown chemopreventive effects in a HBV-related in vitro model. This in vivo study was designed to seek the chemopreventive effects of CLL and its mechanisms. CLL mixture concentrated with dextrose water by boiling was lyophilized. CLL extracts were administrated to HBV X protein (HBx) transgenic mice aged 4 weeks for 2 to 4 weeks and aged 6 months for 3 months. After administration, histological changes in the liver tissue and expression of HBx-related genes were investigated. CLL-treated mice showed less visceral fat, a smaller liver/body weight ratio and delayed liver pathogenesis. Proliferating cell nuclear antigen (PCNA) expression was also increased in CLL-treated HBx transgenic mice, indicating regeneration of damaged liver tissue. CLL treatment decreased expression of HBx and increased p21 and cyclin D1 in livers of HBx transgenic mice. In addition, p-p53 was increased after CLL treatment. These results suggest that CLL can have beneficial effects on the early and late stages of liver pathogenesis, preventing and delaying liver carcinogenesis. This drug should be considered as a potential chemopreventive agent for HBV-related hepatocarcinogenesis.

Integrative Tumor Board: A Case Report and Discussion From East-West Cancer Center

Aim. To introduce the application of wheel balance cancer therapy (WBCT) in cancer treatment through an intensive study of a representative patient. Method. A 59-year-old, female patient who was a secondhand smoker and who carried no significant medical and family history was diagnosed with T1N0M1 (stage 4) lung adenocarcinoma. Immunohistochemistry was positive for thyroid transcription factor-1, CK7, cyclin D1, p53, and Ki-67 on both lung biopsy specimens. An open lobectomy with mediastinal nodal dissection followed. The patient was transferred to East-West Cancer Center (EWCC) suffering from exertional dyspnea (NYHA [New York Heart Association] grade 2) with thoracic pain and pain in both upper limbs because of the decreased ventilatory capacity and reduced exercise capacity after a pulmonary resection. She also complained of emotional stress. EWCC provided WBCT, a multimodality alternative protocol based on the theory of traditional oriental medicine. Physicians regularly monitored whether metastasis or recurrence existed through blood and radiation tests. Observations and dialogues with the patient were used to record the changes in symptoms such as thoracic pain, dyspnea, anxiety, and fear. Results . Treatments at EWCC led to a disease-free survival of 28 months without adjuvant chemotherapy. Neither metastasis nor recurrence occurred during this period. Exertional dyspnea recovered to NYHA grade 1. Other physical and psychological symptoms were alleviated. Conclusion. This report suggests that WBCT may have the potential for extending life expectancy, controlling symptoms, and improving quality of life in cancer patients.