• Title/Summary/Keyword: 2D cell lines

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Immune Stimulating Efficacy of Insoluble $\beta$-l, 3-glucan from Agrobacterium sp. R259 KCTC 10197BP (Agrobacterium sp. R259 KCTC 10197BP로부터 생산된 $\beta$-1, 3-glucan의 면역 활성 효능)

  • 심정현;최원아;상병찬;윤도영
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.459-465
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    • 2002
  • $\beta$-l, 3-glucans are well known to enhance the immune reactions, resulting in antitumor, antibacterial, antiviral, anticoagulatory and wound healing activities. $\beta$-1, 3-glucans have various activities depending on molecular weight, degree of branching, conformation, water-solubility and intermolecular association. However, the $\beta$-1, 3-glucans linked backbone structure is essential and $\beta$-D-glucopyranosyl units are required for immunopotentiating activities. In this study, we tested the immunophamacological activities of insoluble $\beta$-1, 3-glucan from Agrobacterium sp. R259 KCTC 10197BP and confirmed the following activities: (1) IFN-${\gamma}$ production in PBMCs in the presence or in the absence of PHA, LPS, IL-18, and IL-12; (2) the induction of various cytokines in the spleen and thymus; (3) the adjuvant effect on the antibody production; (4) the cytotoxic and antitumor effects on cell lines and ICR mice. These results strongly suggest that $\beta$-1, 3-glucan from Agrobacterium sp. R259 KCTC 10197BP possesses various immunopharmacologica1 activities.

Inhibition of the Replication of Hepatitis C Virus Replicon with Nuclease-Resistant RNA Aptamers

  • Shin, Kyung-Sook;Lim, Jong-Hoon;Kim, Jung-Hye;Myung, Hee-Joon;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1634-1639
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    • 2006
  • Hepatitis C virus (HCV)-encoded nonstructural protein 5B (NS5B) possesses RNA-dependent RNA polymerase activity, which is considered essential for viral proliferation. Thus, HCV NS5B is a good therapeutic target protein for the development of anti-HCV agents. In this study, we isolated two different kinds of nuclease-resistant RNA aptamers with 2'-fluoro pyrimidines against the HCV NS5B from a combinatorial RNA library with 40 nucleotide random sequences, using SELEX technology. The isolated RNA aptamers were observed to specifically and avidly bind the HCV NS5B with an apparent $K_d$ of 5 nM and 18 nM, respectively, in contrast with the original RNA library that hardly bound the target protein. Moreover, these aptamers could partially inhibit RNA synthesis of the HCV subgenomic replicon when transfected into Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic agents of HCV infection but also as a powerful tool for the study of the HCV RNA-dependent RNA polymerase mechanism.

A Study on the Tyrosinase Inhibitory and Antioxidant Effect of Microalgae Extracts

  • Ji, Keunho;Kim, Yeeun;Kim, Young Tae
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.167-173
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    • 2021
  • Reactive oxygen species (ROS) disrupt the cellular redox balance, exert cytotoxic effects, and consequently promote the development of various diseases in humans. Previous studies have reported that antioxidants counteract the adverse effects of ROS. Several studies examine the whitening effects of various agents based on their ability to inhibit tyrosinase activity. Tyrosinase is a critical enzyme involved in the synthesis of melanin, which protects the skin against radiation. Various agents exhibiting antioxidant and tyrosinase inhibitory activities have been synthesized. However, these synthetic drugs are associated with toxicity, decreased safety, and poor skin penetration in vivo, which has limited the clinical application of synthetic drugs. This study examined the antioxidant and tyrosinase inhibitory activities of some microalgae. The methanol, dichloromethane, and ethyl acetate extracts of four microalgal species (Tetraselmis tetrathele, Dunaliella tertiolecta, Platymonas sp., and Chaetoceros simplex) were prepared. The physiological and whitening effects of microalgal extracts were investigated by measuring the antioxidant and tyrosinase inhibitory activities. The ethyl acetate extract of D. tertiolecta exhibited the highest antioxidant and tyrosinase inhibitory activities. Future studies must focus on examining the whitening effects of microalgae on cell lines to facilitate the development of microalga-based therapeutics for skin diseases, functional health foods, and whitening agents. Thus, microalgae have potential applications in the pharmaceutical, food, and cosmetic industries.

Enhanced Production of Oleanolic Acid by the Elicitation in Oldenlandia diffusa Suspension Cell Cultures (백화사설초의 현탁세포배양에서 Elicitation에 의한 Oleanolic acid 생산성 증대)

  • Lee Yong-Il;Kim Dong-Il
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.471-477
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    • 2004
  • Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

Some In-Vitro and In-Vivo Biological Activities of Hot Water Extracts from Fruit Body and Cultured Mycelium of Hericium erinaceum (Hericium erinaceum 균사체와 자실체 열수 추출물의 몇몇 In-Vitro 및 In-Vivo 생물활성)

  • Jung, Jae-Hyun;Lee, Kwang-Ho;Lee, Shin-Young
    • KSBB Journal
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    • v.22 no.1
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    • pp.22-29
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    • 2007
  • The water-soluble materials extracted from fruit bodies and mycelium of H. erinaceum were prepared. In-vitro anticancer activities on cancer cells and In-vivo proliferation effect on mouse peritoneal exudate cell and spleen cell of samples were investigated. Also, nitric oxide (NO) generation of peritoneal exudate cell, IL-2 production capacity of spleen cells and phagocytic activity of peritoneal macrophages were examined. The water extracts of H. erinaceum suppressed the proliferation of cancer cell (HeLa, Raw264.7, Jurkat, KATO3, EL4, LyD9) with concentration-dependent. The water extract from fruit body showed better suppression effect than that from mycelium in most of cancer cells used. The anticancer effect of water extract of fruits body in the range of 0.01 and 10 mg/ml for Raw 264.7 and EL4 cell lines were the same as the Taxol with one thousandth equivalent of fruit body concentration. Water extracts of fruit body and liquid-cultured products of H. erinaceum induced nitric oxide (NO) generation of peritoneal exudate cell and increased NO generation by stimulus of lipopolysaccharide. Water extracts alone did not induce the proliferation and IL-2 production capacity of spleen cells. However, spleen's proliferation and IL-2 production were induced significantly by the addition of lipopolysaccharide and Con A (concanavalin A) or Con A alone, and the effectiveness of mycelium extract with water were more active than those from fruit body.

DNA and Proteomic Analysis of Ginseng Radix Rubra Herbal-acupuncture Solution(GRR-HAS) on Gene Expression in HepG2 Carcinomar Cells (홍삼약침액(紅蔘藥鍼液)의 DNA와 단백질 발현(發顯)에 미치는 영향(影響))

  • Won, Eun-Ju;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul;Lee, Kyung-Min
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.177-190
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    • 2006
  • Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

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Synthesis and Importance of Bulky Aromatic Cap of Novel SAHA Analogs for HDAC Inhibition and Anticancer Activity

  • Chun, Pu-Soon;Kim, Won-Hee;Kim, Jung-Su;Kang, Jin-Ah;Lee, Hye-Jin;Park, Ji-Young;Ahn, Mee-Young;Kim, Hyung-Sik;Moon, Hyung-Ryong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.6
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    • pp.1891-1896
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    • 2011
  • On the basis of potent HDAC-inhibitory activity and anticancer activity of SAHA, novel SAHA derivatives 3a-d and 7 with a bulky cap such as p-dimethylaminophenyl, 4-phenylaminophenyl, 4-phenyloxyphenyl, 9H-fluorenyl or naphthalenyl ring were synthesized starting from the corresponding aryl amines or naphthalenyl acetic acid using an EDC-mediated amide coupling reaction in the presence of HOBt followed by a nucleophilic addition-elimination reaction with hydroxylamine. Compounds 3b, 3c and 3d showed more potent inhibitory activity on total HDACs (14~27-fold), HDAC1 (8~15-fold), HDAC2 (1.3~25-fold) and HDAC7 (1~3-fold) and more potent anticancer activity (2~22-fold) against MCF-7, MDA-MB-231, MCF-7/Dox, MCF-7/Tam, SK-OV-3, LNCaP and PC3 human cancer cell lines than SAHA.

Immunomodulatory Effects of β-sitosterol and Daucosterol Isolated from Dioscorea batatas on LPS-stimulated RAW 264.7 and TK-1 Cells (산약에서 분리한 β-sitosterol과 daucosterol의 RAW 264.7 세포와 TK-1 세포에서의 면역 활성 조절 효능)

  • Park, Min-Kyung;Cho, Sehee;Ahn, Tae-Kyu;Kim, Do-Hyun;Kim, So-Yeon;Lee, Jin-Wook;Kim, Jee-In;Seo, Eul-Won;Son, Kun-Ho;Lim, Jae-Hwan
    • Journal of Life Science
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    • v.30 no.4
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    • pp.359-369
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    • 2020
  • Although many studies on immune modulatory materials have used RAW 264.7 cells, few have used T cell-derived TK-1 cell lines. Moreover, although some studies have investigated the efficacy of plant-derived β-sitosterol, few have examined the immunomodulatory activity of its analogue, daucosterol. In this study, β-sitosterol and daucosterol were isolated from D. batatas and identified by nuclear magnetic resonance spectroscopy. To evaluate the immune-enhancing or inhibitory effects of the isolated phytosterols, the expression levels of the inflammatory response genes COX-2, TNF-α, IL-6, and iNOS were analyzed by RT-PCR. The relative expression levels of TNF-α and iNOS in RAW 264.7 cells were increased more than threefold with β-sitosterol treatment comparing to those of untreated control. In the case of TK-1 cells, the expression level of TNF-α was decreased and the expression level of iNOS was increased in a β-sitosterol concentration-dependent manner. The expression levels of COX-2, TNF-α, and IL-6 increased by approximately 0.7-1.2 times in RAW 264.7 cells treated with daucosterol compared to those of untreated control, but iNOS expression decreased by 0.8-0.18 times. In the case of daucosterol-treated TK-1 cells, the expression levels of TNF-α, IL-6, and iNOS were markedly reduced from those of TK-1 cells treated only with lipopolysaccaride. As a conclusion, β-sitosterol treatment increased TNF-α and iNOS expression levels in RAW 264.7 cells, thus exerting an immune- boosting effect. However, in TK-1 cells, iNOS expression increased while TNF-α expression decreased, indicating an immunosuppressive activity of β-sitosterol. Daucosterol appears to exert an immunosuppressive effect in both macrophages and T cell lines by inhibiting iNOS expression in RAW 264.7 cells and greatly inhibiting the expression of TNF-α, IL-6, and iNOS in TK-1 cells.

Antimutagenic and Cytotoxic Effects of an Ethanol Extract of Buckwheat Sprout (메밀싹 에탄올 추출물의 항돌연변이원성 및 암세포 성장억제 효과)

  • Cui, Cheng-Bi;Lee, Eui-Yong;Ham, Seung-Shi;Lee, Deuk-Sik
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.212-218
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    • 2008
  • This study was performed to determine the antimutagenic and anticancer effects of ethanol extract of buckwheat sprout using Ames test and SRB assay, respectively. An ethyl acetate fraction (200 ${\mu}/plate$) from the ethanol extract of buckwheat sprout showed inhibition rate of 80.6% against the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Salmonella typhimurium TA100 strain. Also the ethyl acetate fraction (200 ${\mu}/plate$) showed higher antimutagenic activity than other fractions against the mutagenesis induced by 4-nitroquinoline-1-oxide (4NQO) in Salmonella typhimurium TA98 and TA100. In addition, the ethyl acetate fraction (200 ${\mu}/plate$) showed high antimutagenic effect of 80.9% and 85.9% against the mutation of TA98 and TA100 strains induced by 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1), respectively. The cytotoxic effects of each solvent fraction from the ethanol extract of buckwheat sprout against human cancer cell lines including lung carcinoma (A549), gastric carcinoma (AGS), breast adenocarcinoma (MCF-7), hepatocellular carcinoma (Hep3B), and colon adenocarcinoma (Colo 205) were investigated. The ethyl acetate fraction of buckwheat sprout ethanol extract at the concentration of 1.0 mg/ml showed strong cytotoxic activities of 70.3, 94.8, 79.6, 82.3, and 73.2% against A549, AGS, MCF-7, Hep3B and Colo 205 cancer cell lines, respectively.

The Antiproliferative Effects of Compounds Isolated from Schisandra chinensis (오미자로부터 분리된 화합물의 암세포 증식 억제 효과)

  • Suh, Won-Se;Park, So Yeon;Min, Byung Sun;Kim, Sea Hyun;Song, Jeong Ho;Shim, Sang Hee
    • Korean Journal of Food Science and Technology
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    • v.46 no.6
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    • pp.665-670
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    • 2014
  • We isolated twelve lignans and three terpenoids were isolated from the n-hexane fraction of Schisandra chinensis extract. Using spectroscopic data and comparison with available literature, the following compounds were identified: (1) wuweizisu C, (2) gomisin N, (3) deoxyschisandrin, (4) gomisin A, (5) schisandrin, (6) chamigrenal, (7) schisanlactone D, (8) methylgomisin O, (9) angeloylgomisin O, (10) (-)-gomisin $L_2$, (11) schisandronic acid, (12) (-)-gomisin $L_1$, (13) (+)-gomisin $K_3$, (14) gomisin J, and (15) tigloylgomisin H. Notably, this was the first finding that compound (8) was isolated from this plant. Each compound was evaluated for its in vitro cytotoxic activities toward HL-60 (human leukemia), HeLa (human cervical carcinoma), and MCF-7 (breast cancer) cell lines. Compounds (7), (8), and (9) exhibited strong cytotoxic effects on HL-60 ($IC_{50}$ 7.37, 6.60, and $8.00{\mu}M$, respectively), whereas compound (6) exhibited weak cytotoxicity towards MCF-7 ($IC_{50}$ $30.50{\mu}M$). In addition, compound (8) showed the strongest activity towards HeLa cells ($IC_{50}$ $1.46{\mu}M$).