• Title/Summary/Keyword: MTT tetrazolium

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Development of Anticancer Agents from Korean Medicinal Plants (Part 4). Antitumor Activity of the Butanol Soluble Fraction of Perilla frutescens (한국산 생약으로부터 항암물질의 개발(제4보) 소엽 부탄올 가용분획의 항암활성)

  • 최규은;곽정숙;김영옥;백승화;한두석
    • Toxicological Research
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    • v.13 no.4
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    • pp.311-316
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    • 1997
  • This study was carried out to develop antitumor effect of the n-butanol soluble fraction of Perilla frutescens on (KB cells) human oral epitheloid carcinoma cells. The cytotoxictty of methanollc extract of Perilla frutescens on KB cells was evaluated by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) assay. The antitumor activity of various fractions obtained from n-butanol soluble fraction of Perilla frutescens was evaluated in human oral epithelold carcinoma cells. The antitumor acavity of the n-butanol soluble fraction on human oral epitheloid carcinoma cells was evaluated by MTT assay of colorimetric method. The light microscopic study was carried out to observe morphological changes of cultured human oral epitheloid carcinoma cells. These results were obtained as follows; 1. The fractions 1,2 and 3 of the n-butanol soluble fraction of Perilla frutescens were shown significant antitumor activities. 2. The number of human oral epitheloid carcinoma cells were decreased and tend to form cell cluster by treatment with fractions 1,2,3 and 4 of the n-butanol soluble fraction of Perilla frutescens. 3. The fraction 1 of the n-butanol soluble fraction of Perllla frutescens showed the highest antitumor activity on Perilla frutescens. It has been selected as a lead fraction for further examinations.

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In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
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    • v.3 no.3
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    • pp.242-244
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    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

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Cytotoxic Effect of Variable Crops (수종의 작물에 대한 세포독성효과)

  • 문형인
    • Korean Journal of Plant Resources
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    • v.10 no.4
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    • pp.346-350
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    • 1997
  • Antineoplastic activities against human gastric and colon carcinoma cell lines were measured in seventeen methanol extracts of seventeen food and medicinal crops using the MTT(3-[4.5-dimethyl thiazol-2-yl)-2.5-diphenyl tetrazolium bromide) method. Three extracts(Hodeum vulgare, Schizandra chinensis and Zea mays)from seventeen tested crops have been reported to have antineoplastic effects. The extracts obtained from remaining fourteen tested crops failed to show significant cytotoxic effect at the concentration of less than $230{\mu}g/ml$(50% inhibitory concentration).

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The Cytotoxic Activity of 3,4,5-Trihydroxybenzoic Acid Methylester and Related Compounds against Skin and Oral Cancer Cell Lines (3,4,5-Trihydroxybenzoic Acid Methylester와 관련 화합물의 피부암 및 구강암 세포주에 대한 세포독성)

  • Lee, Jae-Sug;Han, Du-Seok;Kang, Jeong-Il;Baek, Jong-Min;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.54 no.2
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    • pp.112-121
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    • 2010
  • The cytotoxic activity of 33,4,5-trihydroxybenzoic acid methylester and related compounds on the growth of normal cell lines, human skin melanoma cells and human oral epithelioid cell line were evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis-[2-methoxy-4-nitro-5-sulfo-phenyl]-2-H-tetrazolium-5-caboxanilide (XTT) methods. 3,4,5-Trihydroxybenzoic acid methylester decreased the cell viability of human skin melanoma cells and human oral epithelioid cells shown by the MTT method and the cell adhesion activity of human skin melanoma cells and human oral epithelioid cells shown by the XTT method. In light microscopy, 100 ${\mu}M$ 3,4,5-trihydroxybenzoic acid methylester showed the highest cytotoxic activity. These results suggest that 3,4,5-trihydroxybenzoic acid methylester has a potential anticancer activity.

Carbofuran Cytotoxicity on Rat Fibroblast Cells and Compensation Effects of Phenobarbital Sodium and 3-Methycholanthrene on Its Toxicity (살충제 Carbofuran이 쥐의 NIH3T3 섬유모세포에 끼치는 독성 및 Phenobarbital Sodium과 3-Methylcholanthrene에 의한 보상효과)

  • Han, Seong-Soo;Rim, Yo-Sup
    • Korean Journal of Environmental Agriculture
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    • v.16 no.2
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    • pp.149-155
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    • 1997
  • This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.

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Studies on the Cytotoxicity and Antitumor Activity of Perilla frutescens (소엽의 세포독성 및 항암작용에 관한 연구)

  • Han, Du-Seok;Chung, Boung-Ho;Yoo, Hyeon-Gyeong;Kim, Young-Ok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.25 no.3
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    • pp.249-257
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    • 1994
  • The cytotoxic and antitumor activity of Perilla frutescens extract on cultured 3T3 fibroblast and skin melanoma cells were evaluated by tetrazolium MTT (MTT) and neutral red (NR) colorimetric assay methods. Lactate dehydrogenase activity was also measured. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and skin melanoma cells. The results were as follows: 1. Water and ether extracts showed a significant cytotoxicity in 3T3 fibroblast and all extracts exhibited a significant anti-tumor activity in skin melanoma cells. Methanol, ethyl acetate and ethanol extracts showed low cytotoxic effects, but exhibited a high anti-tumor activity. 2. The MTT absorbance in 3T3 fibroblast was significantly decreased by treatment with ether, water, chloroform and ethanol extracts and skin melanoma cells was significantly decreased by treatment with all extracts. The difference in MTT absorbance in two cell types was most remarkable when treated with methanol and ethanol extracts. 3. Methanol and ethyl acetate extracts showed the strongest effect in growth inhibition of melanoma cells. These results indicated that methanol extract possessed a low cytotoxicity and a strong anti-tumor activity.

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The antiproliferative activity of cannabidiol ethyl ethers against human ora epitheloid carcinoma cells

  • Baek, Seung-Hwa;Kang, Kil-Ung;Chung, Soon-Ryang;Kim, Hyung-Min;Chung, Woo-Young;Han, Du-Seok
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.45-54
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    • 2000
  • Cannabidiol derivatives (1, 2 and 3), and 5-fluorouracil (4, 5-FU) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed a potent inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activity of these compounds (1, 2, 3 and 4) was in a dose-dependent over the micromolar concentration ranges from $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decreases in the following order: CBD > 5-FU > CBDME > CBDDE by the MTT assay and SRB assay. Cannabidiol derivatives (1, 2 and 3), and 5-FU were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were in a dose-dependent over the micromolar concentration range $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 3T3 fibroblasts shows that their susceptibility to these compounds decreases in the following order; CBD > 5-FU > CBDDE > CBDME by MTT assay, CBD > 5-FU > CBDME > CBDDE by SRB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against KB cell lines.

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Effects of Copper / Zinc-Containing Superoxide Dismutase (Cu, Zn-SOD) and Catalase on Paraquat-Induced Injury in Primary Cultured Rat Skin Fibroblast (일차 배양한 백서 피부섬유아세포에서 Paraquat 독성에 미치는 SOD 와 Catalase 의 영향)

  • Cha, Jong Hui;Yu, Ui Gyeong
    • Journal of the Korean Chemical Society
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    • v.38 no.1
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    • pp.74-79
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    • 1994
  • The participation of superoxide in initiating tissue damage derived from xenobiotics is best illustrated by paraquat intoxication. In the present study, the roles of superoxide dismutase and catalase on paraquat-induced cell injury were investigated using primary cultured rat skin fibroblast. The degree of cell injury was assassed by the conversion of reduced MTT to a blue formazan. Paraquat produced concentration-and time-related cell injury in cultured rat skin fibroblast. Paraquat induced-cell injury was aggravated by pretreatment of aminotriazol (AT: 10 mM), an catalase inhibitor, and attenuated by addition of catalase (100∼500 unit/ml). However, the effects of diethyldithiocarbamate (DDC : 10 mM), copper- and zinc-containing superoxide dismutase (Cu, Zn-SOD) inhibitor, and Cu, Zn-SOD on paraquat-induced injury were not significant. These results suggest that hydrogen peroxide might be more responsible factor than superoxide in the pathogenesis of paraquat-induced cell injury.

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CYTOTOXICITY OF RETROGRADE FILLING MATERIALS TESTED BY 51Cr RELEASE, MIT AND LD ACTIVITY (51Cr방출과 MTT 및 LD활성도를 이용한 역충전재의 세포독성에 관한 연구)

  • Choi, La-Young;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.19 no.2
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    • pp.409-428
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    • 1994
  • Endodontic surgery is performed when conventional endodontic therapy fails or is contraindicated. In such cases, retrograde filling materials including amalgam, composite resin, and various cements have been used. Biocompatibilty and margin sealing ability of retrograde filling materials are important for the long term success of endodontic surgery. In vitro cell culture is frequently used as the method of measuring the biocompatibilty of dental materials. The purpose of this study was to evaluate the cytotoxicity of six kinds of retrograde filling materials including newly developed light curing glass ionomer cements. Each material was mixed according to. the manufacture's instruction and evaluated as : freshly mixed, 24-hour after mixing, and 168-hour after mixing respectively. The elution solution was extracted after 24-hour contact with materials using media. Cytotoxicity was evaluated by direct contact, or elution contact. Test results of radiochromium($^{51}Cr$) release, cell viability using tetrazolium dye (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl dimethyltetrazolium bromide(MTT) test and lactate dehydrogenase(LD) of damaged L929 cells were analyzed. In the $^{51}Cr$ release of direct contact, all experimental retrograde filling materials except amalgam and glass ionomer cement showed increased cytotoxicity compared to control. In the $^{51}Cr$ release of elution solution, the released $^{51}Cr$ was so minimal that it was impossible. to evlauate the cytotoxicity exactly. The elution solutions of glass ionomer cement and IRM showed marked cytotoxicity in MTT test. LD enzyme activity was highest in tests of direct contact with composite, light curing composite, and light curing glass ionomer cement and IRM. Amalgam revealed least cytotoxicity while IRM showed cytotoxicity using all three methods. Composite, light curing composite and light curing glass iomomer cement were cytotoxic in the tests of $^{51}Cr$ release and LD activity. Glass ionomer cement showed cytotoxic effect only in the MTT method. From these results it is suggested that the standardization and optimization of cytotoxicity testing, especially using elution solutions, should be strongly advised.

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Screening of Korean Medicinal and Food Plants with Antitumoral Activity (국내 약용 및 식용식물중 항종양활성 식물탐색)

  • Chung, Ill-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk;Ahn, Jong-Seog;Ahn, Soon-Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.37-44
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    • 1999
  • This study was conducted to screen antitumoral activity by in vitro bioassay method using 60 Korean medicinal and food plants extracted by 80% EtOH. Antitumor activity test was applied by the PKC(protein kinase C) and antibleb formation, PLC (Phospholipase C), and colorimetric tetrazolium assay (MTT assay) methods. Chenopodjum album and black Glycine max showed high antitumoral activity by 73.5% and 81.0%, respectively, against PKC by bleb-forming assay and PKC enzyme assay on human chronic leukemia K562 cell. Black Glycine max also showed 91.2% antitumoral activity in the PLC method and the lowest $IC_{50}$ $value(4.7{\mu}g/ml)$ by MTT method against P-338 cell line. In the effect of the concentration treatment on antitumoral test, the more concentration indicated the more activity value.

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