• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.27-34
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• 2000

The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

• Journal of Korean Neurosurgical Society
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• 제38권2호
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• pp.126-131
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• 2005

Objective : The choice of tumor antigen for dendritic cell[DC]-loading has still been an unresolved problem in the DC-based vaccine strategies against malignant gliomas that has not been found well-characterized tumor specific antigens. In this study, we compare tumor-specific T cell response induced by glioma apoptotic body[GAB]-pulsed DCs to response induced by glioma cell lysate-pulsed ones quantitatively. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor and interleukin[IL]-4 from peripheral blood mononuclear cells[PBMCs] of HLA-A2 positive healthy donors were cultured. Each GABs and glioma cell lysate generated from HLA-A2 positive T98G glioblastoma cells were co-incubated with DCs. $CD8^+$ T lymphocytes isolated from PBMCs of same donors were cultured in media containing IL-2 and either stimulated by GAB- or lysate-pulsed DCs three times at a weekly interval. The interferon[IFN]-${\gamma}$ concentrations of each cell culture supernate were measured by enzyme immunoassay technique. Cytolytic activity of the generated cytotoxic $CD8^+$ T cells either stimulated with GAB- or lysate-pulsed DCs was determined by a standard 4-h $^{51}Cr$-release assay. Results : IFN-${\gamma}$ production and cytolytic activity of effector T cells stimulated by GAB-pulsed DCs were significantly higher than those of T cells stimulated by lysate-pulsed ones. Conclusion : These results indicate the choice of antigen is a critical determinant in the induction of antitumor immunity against malignant glioma. Antigen preparations from GABs represent a promising alternative to glioma cell lysate in DC-based glioma vaccine strategies.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.3035-3042
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• 2015

Background: Isorhamnetin (Iso), a novel and essential monomer derived from total flavones of Hippophae rhamnoides that has long been used as a traditional Chinese medicine for angina pectoris and acute myocardial infarction, has also shown a spectrum of antitumor activity. However, little is known about the mechanisms of action Iso on cancer cells. Objectives: To investigate the effects of Iso on A549 lung cancer cells and underlying mechanisms. Materials and Methods: A549 cells were treated with $10{\sim}320{\mu}g/ml$ Iso. Their morphological and cellular characteristics were assessed by light and electronic microscopy. Growth inhibition was analyzed by MTT, clonogenic and growth curve assays. Apoptotic characteristics of cells were determined by flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay, immunocytochemistry and terminal deoxynucleotidyl transferase nick end labeling (TUNEL). Tumor models were setup by transplanting Lewis lung carcinoma cells into C57BL/6 mice, and the weights and sizes of tumors were measured. Results: Iso markedly inhibited the growth of A549 cells with induction of apoptotic changes. Iso at $20{\mu}g/ml$, could induce A549 cell apoptosis, up-regulate the expression of apoptosis genes Bax, Caspase-3 and P53, and down-regulate the expression of Bcl-2, cyclinD1 and PCNA protein. The tumors in tumor-bearing mice treated with Iso were significantly smaller than in the control group. The results of apoptosis-related genes, PCNA, cyclinD1 and other protein expression levels of transplanted Lewis cells were the same as those of A549 cells in vitro. Conclusions: Iso, a natural single compound isolated from total flavones, has antiproliferative activity against lung cancer in vitro and in vivo. Its mechanisms of action may involve apoptosis of cells induced by down-regulation of oncogenes and up-regulation of apoptotic genes.

• 대한바이러스학회지
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• 제28권1호
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• pp.71-78
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• 1998

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

• Advances in Traditional Medicine
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• 제1권1호
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• pp.37-44
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• 2000

In the previous paper (Kim et al., Glycoconjugate Journal 16, 247-252, 1999), heteropolysaccharides from Korean medicinal plant, Phellodendri cortex (Hwangbek) showed a poten B-Iymphocyte-stimulating activity in a system using polyclonal antibody forming cells in C57BL/6XC3H mice at dosages of 2-10 mg. In a series of biolgical active polysaccharides from natural medicinal plants, the polysaccharide fractions were isolated and purified from Phellodendron chinese SCHNEID, and antitumor activities were examined at dosages of 2, 5 and 10 mg/100 g. F-7 and F-8 showed the highest tumor inhibitory activities (inhibition ratio 96.4 and 98.2% in 2 mg/100 g), and in dose of 5 mg/100 g, the inhibitory ratios were 95.3 and 97.5%, respectively. Furthermore, 10 mg/100 g of intraperitoneal (i.p.) injection gave 97.3 and 98.7% of inhibition. In oral administration, the inhibitory activities were not markedly observed, indicating that the polysaccharides are directly acting to immune system. When the effects on TS and TK activities were determined, TS activities in the F-2 and F-7-treated mice were markedly suppressed to 73.7% and 79.5% of that in the control (p<0.01), while there was little difference in TK activity with a slight decrease in F-2 only. However, in i.p. injection, TS activities in the F-2, F-5, F-7 and F-8-treated mice were markedly suppressed to 83% to 85% of that in the control (p<0.01). Furthermore, there was also significant differences in TK activities in F-2, F-5, F-7 and F-8-treated mice (p<0.05). Therefore, polysaccharide fraction F-8 was further purified to active fractions of F-9 and F-11 by gel permeation chromatography using TSK Gel HW50S. The purified polysaccharides of F-9 and F-11 were composed of GlcNAc (47.3%), Gal (24.7%) and Man (28.0%). These results clearly indicated that the i.p. injection is much effective to suppress tumor growth than oral administration.

• 한국미생물·생명공학회지
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• 제41권3호
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• pp.350-357
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• 2013

지의류는 사막에서 북극지방까지 이르는 극한 환경에서도 생존 가능한 곰팡이, 조류 또는 시아노박테리아 등으로 구성된 공생체이다. 몇몇 지의류 공생체들은 항균, 항곰팡이, 항바이러스, 항암, 항산화 및 항염증 등과 같은 많은 생물학적 활성을 지닌 넓은 범위의 이차대사물질을 생산한다. 지의류와 공생 관계인 박테리아에 관하여는 아주 일부 알려져 있다. 최근 본 연구팀은 북극 지의류 Stereocaulon spp로부터 4종류의 미생물을 분리하였으며, DPPH와 ABTS 측정법을 이용하여 그들의 항산화능을 조사하였다. 또한 총 폴리페놀 함량과 총 플라보노이드 함량 분석 등도 측정되었다. 강력한 라디컬 소거능은 지의류 추출물을 이용하여 수행하였다. 본 연구에서 조사된 4종류 중, Bosea vestrisii 36546(T)의 에틸아세테이트 추출액은 DPPH 분석에서 86.8% 그리고 ABTS 분석에서 75.2%에 달하는 억제력과 함께 가장 강력한 자유 라디컬 소거능을 보여주었다. 따라서 이러한 결과들로부터 지의류 유래 박테리아 종들이 천연 항산화제로서 잠재적인 소재가 될 수 있다는 것을 제안한다.

• 한국식품과학회지
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• 제30권5호
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• pp.1236-1242
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• 1998

한국산 표고버섯 산련 1호를 C/N비가 13.1인 버섯 완전액체배지에서 배양하여 균사체를 증식시킨 후 생리활성을 보이는 단백다당체를 추출하였다. 표고버섯 균사체 액체배양시 생성되는 단백다당체는 균사체 세포벽에 약 80%가 존재하고, 약 20%정도는 세포밖으로 분비하는 것으로 나타나 단백다당체의 추출은 균사체가 함유된 배양액 전체를 이용하여 추출하는 것이 높은 수율을 얻을 수 있었다. 그리고 균사체로부터 단백다당체의 추출방법으로 열수추출 및 glass bead mill로 세포벽을 파쇄한 후 열수추출, cellulase 처리 후 열수추출을 비교한 결과 물리적으로 세포벽을 파쇄한 후 60분간 열수추출하는 것이 배양액 100 mL당 약 930 mg의 조단백다당체가 추출되어 수율이 가장 높았다. 추출한 조단백다당체를 사용하여 protease 처리, 그리고 DEAE-cellulose 및 Sephadex G-100 column chromatography를 통하여 단백다당체를 정제한 후 단계별 시료에 대한 mouse leukemic cell인 $P_{388}$의 증식억제는 53.7, 62.2, 93.7, 97.4%로 정제도가 높아짐에 따라 증가하는 것으로 나타났다. Sephadex G-100 column으로 처리한 정제 단백다당체를 TLC/FID로 분석한 결과 단일 peak로 나타나 순수물질임을 확인하였다. 그리고 단백다당체의 성분을 분석한 결과 다당함량은 46.1%로 구성단당류는 glucose 및 galactose, xylose, mannose로 구성되어 있었고, 단백질은 약 7.3%로 주로 proline 및 glycine의 함량이 높았으며 methionine 및 leucine은 거의 없는 것으로 밝혀졌다. 그외에 무기질로서 Na, K, Zn, Ca 등이 존재하는 것으로 나타났다.

• 한국잡초학회지
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• 제14권1호
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• pp.16-22
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• 1994

경기도 광주군 및 연천군 일원에서 채취한 93종의 잡초종(雜草種)을 대상으로 항균활성(抗園活性), 항종양활성(抗腫瘍活性), 항산화활성(抗酸化活性), 제초활성(除草活性) 등 몇가지 생리활성(生理活性)을 조사함으로써 제초(雜草)의 유용적(有用的)인 측면(則面)의 확인 뿐 아니라 나아가서 잡초(雜草)로부터 새로운 생리활성물질탐색(生理活性物質探索)의 가능성(可能性)을 제시하고자 하였다. 곰팡이에 있어서 사과나무 점무늬낙엽병균(落葉病菌)에 대해서는 새삼, 고추 역병균(疫病菌)에 대해서는 돼지풀과 쥐손이풀, 참깨 역병균(疫病菌)에 대해서는 쑥부쟁이와 미국쑥부쟁이가 항균활성(抗菌活性)을 나타내었다. 세균에 있어서는 돼지풀, 쑥, 사철쑥, 여뀌바늘, 구절초, 쥐손이풀, 미국가막사리 등이 비교적 항균활성(抗菌活性)의 범위(範圍)가 넓었으며 개비름, 비름, 망초, 물질경이, 쇠무릎, 물봉선, 벼룩나물 등이 Gram 양성세균인 Bacillus subtilis PCI219에 대하여 선택적(選擇的)인 항균활성(抗菌活性)을 나타내었다. 그리고 새삼, 개가장, 갈대, 쉽사리, 기름나물 등이 세포벽(細胞壁) 결손(缺損) 변이주(變異洙)인 Pseudomonas aeruginosa N10 균주에 대하여 선택적(選擇的)인 항균활성(抗菌活性)을 나타내었다. K562 세포(細胞)에 대한 protein kinase C 활성(活性)으로 조사한 antibleb 활성(活性)은 큰개여뀌, 사철쑥, 국화방망이, 개가장, 쥐손이풀, 바늘사초 등에서 나타났으며 특히 큰개여뀌에서는 조시료(粗試料) 상태에서 10mg/ml 농도(濃度)에서도 강한 활성(活性)을 나타내었다. 항산화활성(抗酸化活性)에 있어서는 바디나물, 쇠뜨기, 승마, 큰개여뀌, 쥐손이풀, 달맞이꽃, 방동사니대가리 등이 비교적 강한 활성(活性을) 나타내었으며 제초활성(除草活性)에 있어서는 여뀌바늘, 기름나물 등이 비교적 강한 활성(活性)을 나타내었다.

• 대한한방종양학회지
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• 제8권1호
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• pp.103-125
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• 2002

This experimental study was carried out to evaluate the anti-tumor and the immunomodulatory effects of Jiaweicitaowan(加未慈桃丸) against cancer. The in vitro anti-tumor effects were evaluated by MTT assay. The cytotoxicity, extension of survival days, the effect of inhibition solid tumor which was induced sarcoma 180, and the changes of body weight were evaluated for in vivo effects of anti-tumor. To evaluate the immunomodulatory effects of Jiawei- citaowan(加未慈桃丸), delayed type hypersensitivity, hemagglutinin, hemolysin titers for humoral immune response, rosette forming cells for cell-mediated immune response, natural killer cell activity, proliferation of lymphocyte, productivty of Interleukin-2, and carbon clearance were measured with methotrexate treated mice. The results were as follows; 1. In the case of existence ability of tumor cell, IC50 had an anti-tumor ativity resulted 2.52mg/ml to SNU-C4. 0.41mg/ml to SNU-396, resulted to 0.09mg/mlSNU-1. 2. The groups of Jiaweicitaowan(加未慈桃丸) 10mg/ml, 20mg/kg had no body weight loss. reduction in intake of water and feed, so these had no toxicity. 3. In the case of the effect of extention of existence. the group of 20mg/kg Jiaweicitaowan(加未慈桃丸) extract treated group was showed 250% in ILS. 4. The effect of inhibition solid tumor was significantly decreased in both 10mg/kg, 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group S. The groups of 10mg/kg, 20mg/kg Jiaweicitaowan(加未慈桃丸) had significant effect of body weight change compared to control group. 6. Delayed type hypersensitivity was not significant in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 7. Hemagglutinin and Hemolysin titers were significantly increased by dose-dependent. so these results showed that the humoral immume respose was activated. 8. For the effect of rosette formimg cells was not significant in hoth Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 9. Natural killer cell activity was significantly increased in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group in the ratio of 100: 1, 50: 1 of effector and target cells, but in the ratio of 10:1, the Jiaweicitaowan(加未慈桃丸) extract treated groups were not significant. 10. The proliferation of lymphocyte and productivty of Interleukin-2 were significantly increased by dose-dependent in both 10mg/ kg, 20mg/ kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 11. In the phagocytic effect, the 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated group showed the increasing effect with significance as compared with control group. According to the results, we can suggest that Jiaweicitaowan(加未慈桃丸) has the antitumor and the immunomodulatory effects.

• 대한한의학회지
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• 제17권1호
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• pp.111-131
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• 1996

For the development of antimetastatic agent 41 kinds of crude drugs were used for the evaluation of inhibitory effect of several crude drugs on cell adhesion of pulmonary cancer cells and platelet aggregation. Results were obtained as follows: 1. Water extracts of crude drugs inhibited cell adhesion of A549 to complex extracelluar matrix over 40 % of contol were Houttuyniae Herba, Mylabris, Rhei Radix et Rhizoma, Meliae Cortex, Ferula Resina, Oldenlandiae diffusae Herba at the higher concentration of $10^{-3}g／ml$ while those inhibiting cell adhesion of Bl6-Fo over 40 % of control were $10^{-5}g／ml$ of Houttuyniae Herba, Aurantii Fructus, Lithospermi Radix, Zedoariae Rhizoma. Prunellae Spica, Foeniculi Fructus, Rbei Radix, Scutellariae Radix, Meliae Cortex, Ferula Resina and Oldenlandiae diffusae Herba. 2. MeOH extracts of crude drugs at the concentration of $4{\times}10^{-4}g／ml$ inhibiting cell adhesion of A549 specifically to single extracelluar matrix over 40 % of control were Lithospermi Radix, Agrimoniae Herba, Rhei Radix and Ferula Resina to collagen I, Houttuyniae Herba, Lithospermi Radix, Bupleuri Radix, Salviae miltiorrhizae Radix, Orostachys Herba, Sappan Lignum, Meliae cortex ferula Resina and Coicis Semen to collagen Ⅳ, Mylabris, Agrimoniae Herba to laminin, Houttuyniae Herba and Meliae Cortex to fibronectin. 3. NeOH extracts of crude drugs at the concentration of $4{\times}10^{-4}g／ml$ inhibiting cell adhesion of B16-Fo specifically to single extracelluar matrix over 60 % of control were Lithospermi Radix, Salviae miltiorrhizae Radix, Meliae Cortex and Ferula Resina to collagen I, Lithospermi Radix, Bupleun Radix, Saiviae miltiorrhizae Radix, Ferula Resina and Acanthopanacis Cortex to collagen Ⅳ, Bupleuri Radix, Orostachys Herba to laminin, Houttuyniae Herba to fibronectin. 4. MeOH extracts of crude drugs inhibiting platelet aggregation over 40% of ADP control were at the concentration of $50{\mu}g/m{\ell}$ of Houttuyniae Herba, Angilicae gigantis Radix, Zedoariae Rhizoma. Coicis Semen and $100{\mu}g/m{\ell}$ of Ferula Resina, Orostachys Herba, Salviae miltiorrhizae Radix, Curcumac Radix, Carthami Flos, Lithospermi Radix, Gleditsiae Spina, Sappan Lignum, Acanthopanacis Cortex. These results suggest that several crude drugs including Ferula Resina, Houttuyniae Herba, Lithospermi Radix and Salviae miltiorrhizae Radix chiefly have more possibility to exert antimetastatic activity and require in vivo antimetastatic study.