• Title/Summary/Keyword: C100 cells

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Cytotoxicity of water extract of Dangkwieumja ka Sumsoo on A43l Cells (當歸飮子加蟾수가 皮膚癌細胞(A431)의 細胞毒性에 미치는 影響)

  • Choi, Jeong-Hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.9 no.1
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    • pp.1-15
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    • 1996
  • The purpose of this research was to investigate effect of water extract of DangKwi-Eum-Ja ka Sumsoo(DESE) on the cytotoxicity of human epidemloid cell, A431 cells. The effects of DESE on the proliferation of A431 cells, Balb/c 3T3 cells, mouse thymocytes and splenocnes were estimated by MTT colorimetric assay, and nitric oxide production from mouse peritoneal macrophage was estimated by Griess method. DESE inhibited the proliferation of A431 cells at $10{\mu}g/ml$, and did not affect the proliferation of Balb/c 3T3 cells. DESE decreased the cytotoxicity of mitomycin C or cisplatin on A431 cells, increased the cytotoxicity of mitomycin C or cisplatin on Balb/c 3T3 cells. DESE inhibited the proliferation of mouse thymocytes and splenocytes at $100{\mu}g/ml$. DESE did not affect the nitric oxide production from mouse peritoneal macrophage in vitro, but decreased the nitric oxide production from DESE-treated mouse peritoneal macrophage.

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Multidrug-resistance reversing activity of the local Citrus fruits in Jeju Island, Korea

  • Lee, Sook-Young;Kim, Sun-Min;Hwang, Eun-Ju
    • Proceedings of the Plant Resources Society of Korea Conference
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    • v.18 no.1
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    • pp.41-51
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    • 2005
  • We examined whether extracts from 14 local citrus spp. on Jeju Island (Korea) contained chemosensitizing activity that would increase the cytotoxic effect of vincristine(VCR) in drug-resistant cancer cells. We report that methanol extracts from fruits and flowers of some species had a chemosensitizing effect that reversed P-glycoprotein (Pgp)-mediated multidrug resistance (MDR). Using drug-sensitive AML-2/WT and drug-resistant AML-2/D100 in the absence of VCR in human acute myelogenous leukemia cells we found that fruit or flower extracts alone generally had low cytotoxicity $(IC_{50}>200\;{\mu}g/ml)$. In studies examining the effect of extracts on 120 ng/ml VCR cytotoxicity in drug-resistant AML-2/D100 cells, we found that immature fruit extracts had greater chemosensitizing activity than either extracts from mature fruit or flower. Of the 14 species examined, the immature fruit extract from Inchangkyool (Citrus ichangiensis) showed the hishest chemosensitizing index(CI) valus. Immature fruit extracts of Hongkyool(C. tachibana), Byungkyool(C. platymamma), Cheongkyool(C. nippokoreana) and Jinkyool (C. sunki) also strongly potentiated VCR cytotoxicity in AML-2/D100 cells. The chemosensitizing effect of peel extracts was 2-10-fold that of whole fruit extracts from Hongkyool (C. tachibana), Byungkyool (C. platymamma) and Inchangkyool (C. inchangiensis). The CI values for flower extracts were higher than those for mature fruit extracts, but lower than those for immature fruit extracts. These results indicate that immature citrus fruits contain compounds that do not exert their activity solely through cytotoxicity. In particular, Incahngkyool (C. inchangiensis), Byungkyool(C.platymamma), Cheongkyool(C. nippokoreana) and Hongkyool (C. tahibana) may be useful sources of chemosensitizing compounds.

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Effects of the Protein Fraction of Panax ginseng on Primary Cultured Chicken Skeletal Muscle Cells (인삼 단백분획물이 일차배양한 계배의 근육세포에 미치는 영향)

  • Park, Mi-Jung;Song, Jin-Ho;Lee, Heun-Pa;Kim, Young-Choong
    • Korean Journal of Pharmacognosy
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    • v.21 no.3
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    • pp.210-216
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    • 1990
  • Effects of the protein fraction of Panax ginseng on chicken embryonic skeletal muscle cells cultured with a decfiient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 dalton(fraction B), between 1,000 and 5,000 dalton(fraction C), between 500 and 1,000 dalton(fraction D). According to the microscopic observation, all four protein fractions at the concentration of $10{\sim}100{\;}{\mu}g/ml$ showed the tendency to stimulate the growth and differentiation of the muscle cells. The activity of acetylcholinesterase in muscle cells was significantly elevated by the protein fraction A at the concentration of $100{\mu}{\;}g/ml$. Protein fractions B,C and D significantly enhanced the synthesis of RNA in the muscle cells. The synthesis of DNA in muscle cells was significantly enhanced by protein fractions A,B and C.

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Hexose Uptake and Kinetic Properties of the Endogenous Sugar Transporter(s) in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.327-332
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    • 2005
  • Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenoussugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dG1c). In order to obtain a true measure of the initial rate of uptake, the uptake of $[^3H]2dGlc$ from both low $(100{\mu}M)$ and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1min time course would reflect initial rates of the jexpse uptake. To determine $K_m\;and\;V_{max}$ of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dG1c was measured over a range of substrate concentrations $(50{\mu}M\~10mM)$ 2dG1c uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system for 2dG1c was relatively high, the $K_m$ value for uptake being < 0.45 mM. The $V_{max}$ value obtained for 2dG1c transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

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Role of cAMP, EGF, IGF-I and Protein Phosphorylation in Mammary Development I. Effect of EGF, IGF-I and Photoreactive Cyclic AMP on DNA Synthesis of Mammary Epithelial Cell (유선발달에 있어서 cAMP, EGF, IGF-I 및 단백질 인산화 작용의 역할 I. EGF, IGF-I 및 Photoreactive Cyclic AMP가 유선상피세포의 DNA합성에 미치는 효과)

  • 여인서;박춘근;홍병주
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.49-56
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    • 1993
  • Mouse mammary epithelial cells(NMuMG) were plated onto 24 well phates(100,000 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, EGF)0~100ng/ml) was added simultaneously with IGF-I(10ng/ml), 1$\mu$M photoreactive cAMP(4,5-dimethoxy-2-nitrobenzyl adenosine-3',5' cyclic monophosphate, DMNB) or IGF-I plus DMNB. After 2 hours, the cells were expposed to UV light(300nm, 3 second pulse0 in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml, 18~19 hours after UV exposure). Without IGF-I or DMNB, EGF(10 or 100ng/ml) increased DNA synthesis from 8,362 dpm/well in control to 16,345 or 18,684 dpm/well with EGF(pooled SE=1,239 dpm/well, P<0.05). IGF-I or IGF-I plus DMNB alone increased DNA synthesis from 8,362 dpm/well in control to 17,307 or 20,427 dpm/well, respectively(P<0.05). Addition of IGF-I, DMNB or IGF-I plus DMNB into 0~100ng/ml EGF did not significantly change the shape of dose response curve of EGF alone. In other experiment, EGF or IGF-I plus DMNB into 10ng/ml EGF group exhibited interaction effect in DNAsynthesis [EGF(10ng/ml)=18,497; IGF-I+EGF=22,837; DMNB+EGF=20,658 ; IGF-I+DMNB+EGF=29,658, pooled SE=1,055, P<0.05]. These results indicate that simultaneous activation of EGF, IGF-I and intracellular cAMP interact in DNA synthesis of mouse mammary epithelial cells.

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Red Ginseng Extract Reduced Metastasis of Colon Cancer Cells In Vitro and In Vivo

  • Seo, Eun-Young;Kim, Woo-Kyoung
    • Journal of Ginseng Research
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    • v.35 no.3
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    • pp.315-324
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    • 2011
  • This study investigated the effect of red ginseng extract on metastasis of colon cancer cells in vitro and in vivo. Wound healing migration, cell motility, invasion, and activity, protein expression, and mRNA expression of matrix metalloproteinases (MMPs) were examined in SW480 human colon cancer cells. SW480 cells were cultured with or without $100{\mu}g/L$ PMA in the absence or presence of various concentrations (100, 200, or $300{\mu}g/mL$) of red ginseng extract. Red ginseng extract treatment caused signifi cant suppression of cell motility and invasion (p<0.05) in SW480 cells. Red ginseng extract inhibited MMP-2 and MMP-9 activity and their protein and mRNA expression in a dose-dependent manner (p<0.05) in SW480 cells. For experimental metastasis, BALB/c mice were injected intravenously with CT-26 mouse colon cancer cells in the tail vein, and were orally administered various concentrations (0, 75, 150, or 300 mg/kg body weight) of red ginseng extract for 3 weeks. Numbers of pulmonary nodules were signifi cantly decreased in mice that were fed red ginseng extract (p<0.05). Plasma MMP-2 and MMP-9 activity signifi cantly decreased in response to treatment with red ginseng extract in mice (p<0.05). These data suggest that red ginseng extract may be useful for prevention of cancer invasion and metastasis through inhibition of MMP-2 and MMP-9 pathways.

The Development of Functional Beverage from the Inner Skin of Chestnut Castanea crenata ( In ) Effects on the Regional Cerebral Blood Flow, Mean Arterial Blood Pressure, Proliferation of Thymocytes - (밤 속껍질로부터 기능성 음료의 개발(III) -뇌혈류역학, 평균혈압, 흉선세포 증식율에 미치는 효과-)

  • 정현우;박철훈;전병관
    • The Korean Journal of Food And Nutrition
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    • v.14 no.5
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    • pp.397-404
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    • 2001
  • The purpose of this study was to investigate effects of inner skin of chestnut on the activation of a living body's function (regional cerebral blood flow and mean arterial blood pressure in Sprague-Dawley rats, proliferation of thymocytes in normal mice and L1210 cells transplanted mice) . We used inner skin of chestnut extract(Sample A : inner skin of chestnut-panbroiled after driedextract (100$\^{C}$ ), Sample B , inner skin of chestnut-panbroiled-extract(100$\^{C}$ ) , Sample C : inner skin of chestnut -panbroiled after dried-extract(80$\^{C}$ ), Sample D : inner skin of chestnut-panbroiled-extract(80$\^{C}$)} Regional cerebral blood flow(rCBF) and Mean arterial blood pressure(MABP) were tested using Leser -Doppler Flowmetry(LDF), and the proliferation of thymcytes was tested using a colorimetric tetrazoliun assay ( MTT assay) The experimental results as follows 1. rCBF was significantly increased by Sample C in a dose-dependent manner. 2. MABP was not changed by Sample C in a 0.1mg/kg∼10.0mg/kg treated group. 3. Proliferation of thymocytes was not changed by Sample C in normal mice. 4. Proliferation of thymocytes was significantly accelerated by Sample C in L1210 cells transplanted mice.

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Mitochondrial Dynamics in Red Algae. 3. Filament Apices in Colaconemacaespitosum (Acrochaetiales) and Antithamnion cruciatum (Ceramiales)

  • Garbary, David J.;Zuchang, Pei
    • ALGAE
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    • v.21 no.3
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    • pp.323-332
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    • 2006
  • Mitochondrial distribution and abundance were assessed during the growth of apical and subapical cells in the red algae Colaconema caespitosum (J. Agardh) Jackelman, Stegenga and Bolton and Antithamnion cruciatum (C. Agardh) Nägeli after staining with 3,3’-dihexyloxacarbocyanine iodide [DiOC6(3)] and 2,4’-dimethylaminostyryl-Nethylpyridinium iodide (DASPEI). In fully elongate apical cells of C. caespitosum there were 100-120 mitochondria. During apical cell enlargement and division there is a doubling and then halving of the mitochondrial numbers. Apical cells prior to cytokinesis in young filaments are smaller than in mature filaments (ca. 50 and 100 μm long, respectively) and have fewer mitochondria (ca. 100 and 120 mitochondria per cell, respectively). In older vegetative cells mitochondria tend to aggregate at opposite ends of the cells with some mitochondria associated with the central nucleus or at points of apparent branch initiation. There is a greater density of mitochondria in apical cells of smaller versus larger plants (one mitochondrion per 6.3 μm3 and 9.8 μm3, respectively), suggesting that apical cells of younger plants may be more metabolically active. Male and female gametophytic thalli of Antithamnion cruciatum had similar numbers of mitochondria in apical cells of indeterminate axes, as did gametophytic and sporophytic thalli. There were about 40-50 mitochondria in fully elongated apical cells with about half this number in newly divided apical and subapical cells. Apical cells of determinate branches had more mitochondria (60-77) than indeterminate branches (60-70 vs. 40-50). In both species and in all cell types mitochondrial numbers were highly correlated with cell size.

Effects of Kisspeptin-10 on Lipid Metabolism in Cultured Chicken Hepatocytes

  • Wu, J.;Fu, W.;Huang, Y.;Ni, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1229-1236
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    • 2012
  • Our previous studies showed that kisspeptin-10 (Kp-10) injected in vivo can markedly increase lipid anabolism in liver of quails. In order to investigate the direct effect of Kp-10 on lipid metabolism of hepatocytes in birds, cells were separated from embryos livers and cultured in vitro with 0, 100 and 1,000 nM Kp-10, respectively. The results showed that after 24 h treatment, cells viability was not affected by 100 nM Kp-10, but showed a mild decrease with 1,000 nM Kp-10 compared to the control cells. Based on the results of the cell viability, 100 nM dosage of Kp-10 was selected for the further study and analysis. Compared with control cells, total cholesterol (Tch) contents in 100 nM treated cells were increased by 51.23%, but did not reach statistical significance, while the level of triglyceride (TG), high density of lipoprotein-cholesterol (HDL-C) and low density of lipoprotein-cholesterol (LDL-C) were significantly increased. Real-time PCR results showed that ApoVLDL-II mRNA expression had a tendency to increase, genes including sterol regulatory element-binding protein-1 (SREBP-1), acetyl coenzyme A carboxylase ${\alpha}$ ($ACC{\alpha}$), carnitine palmitoyltransferase 1 (CPT1), 3-hydroxyl-3-methylglutaryl-coenzyme A reductases (HMGCR) and stearyl coenzyme A dehydrogenase-1 (SCD1) mRNA in hepatocytes were significantly down-regulated by 100 nM Kp-10. However, contrary to its gene expression, SREBP-1 protein expression was significantly up-regulated by 100 nM Kp-10. Some of the significant correlations in mRNA expression were found between genes encoding hepatic factors or enzymes involved in lipid metabolism in liver of birds. These results indicate that Kp-10 stimulates lipid synthesis directly in primary cultured hepatocytes of chickens.

THE MOST APPROPRIATE ANTIMITOTIC TREATMENT OF ARA-C IN SCHWANN CELL-ENRICHED CULTURE FROM DORSAL ROOT GANGLIA OF NEW BORN RAT (신생 백서 척수후근절의 슈반세포 배양을 위한 Ara-C 분열억제제의 최적 효과에 대한 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Ahn, Kang-Min;Kim, Nam-Yeol;Sung, Mi-Ae;Hwang, Soon-Jeong;Kim, Ji-Hyuck;Jahng, Jeong-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.2
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    • pp.100-107
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    • 2004
  • Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.