• Title/Summary/Keyword: Cell assay

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Effects of Sopunghwalhyul-tang Water Extract against Xanthine Oxidase / Hypoxanthine(XO/HX)-Induced Neurotoxicity in the Cultured Mouse Spinal Sensory Neurons (소풍활혈탕 열탕액이 XO/HX에 의해 손상된 배양 척수감각신경세포에 미치는 영향)

  • 양경석;신선호
    • The Journal of Korean Medicine
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    • v.21 no.1
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    • pp.29-39
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    • 2000
  • In order to elucidate the toxic mechanism of oxygen radicals in cultured mouse spinal sensory neurons, cytotoxic effect of oxygen radicals was evaluated by M1T assay and NR assay. In addition, protective effect of Sopunghwalhyultang(SPHHT) water extract on oxidant-induced neurotoxicity was investigated on these cultures. Spinal sensory neurons derived from mice were cultured in mediums containing various concentrations of Xanthine Oxidase / Hypoxanthine(XO/HX). Cell viability was measured by MTT assay and NR assay. XO/HX-mediated oxygen radicals remarkably decreased cell viability of cultured spinal sensory neurons in a dose-and time-dependent manner. And also, SPHHT blocked XO/HX-induced neurotoxicity in these cultures. These results suggest that oxygen radicals are toxic and SPHHT are effective in blocking against the oxidant-induced neurotoxicity in cultures of spinal sensory neurons of mice.

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Pre-validation of Colony Forming Efficiency Assay for Assessing the Cytotoxicity of Nanomaterials (나노물질의 세포독성 평가법으로 Colony Forming Efficiency Assay에 대한 검증연구)

  • Jo, Eunhye;Lee, Jaewoo;Park, Sun-Young;Kim, Pilje;Choi, Kyunghee;Eom, Igchun
    • Journal of Environmental Health Sciences
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    • v.41 no.1
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    • pp.17-23
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    • 2015
  • Objectives: The cytotoxcities of Au, Ag, SWCNT, $SiO_2$, and ZnO nanomaterials were evaluated in order to assess their potential toxicological effects in in vitro cell models using colony forming efficiency (CFE) assay. Methods: The CFE assay of the test materials was carried out on Hep G2 cells. The size distribution of nanomaterials was studied by transmission electron microscopy (TEM). Changes in cell viability after treatment with a toxicant will result in a decreased number of colonies formed in comparison to solvent. Results: The TEM images show that all the particles except SWCNT and ZnO can be considered approximately spherical. The gold and $SiO_2$ nanoparticles show no response (no toxicity) in concentration response experiments. A statistically significant toxic effect was found in Hep G2 cells treated with Ag, SWCNT and ZnO nanomaterials. Conclusion: In this study, we considered CFE assay to be a promising test for screening studies for cytotoxicity with physicochemical analysis.

Development of Anticancer Agents from Korean Medicinal Plants. Part 7. Cytotoxic Activity of the Chloroform soluble Fraction of Perrila frutescens Against Human Oral Epitheloid Carcinoma Cells (한국산 생약으로 부터 항암물질의 개발 (제7보), 소엽의 Chloroform 가용성 분획이 인체 구강유상피암종세포에 미치는 세포독성작용)

  • Han, Du-Seok;Kim, Young-Il;Choi, Kyw-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.37-42
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

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Gold Nanoparticles Inhibit AGEs Induced Migration and Invasion in Bovine Retinal Endothelial Cells (소망막내피세포에서 금 나노입자의 최종당화산물에 의한 세포 이동 및 침윤성 억제 효과)

  • Chae, Soo-Chul
    • Korean Journal of Environmental Biology
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    • v.28 no.1
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    • pp.8-13
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    • 2010
  • This study aimed the role of gold nanoparticles (AuNP) in advanced glycation end-products (AGEs) induced migration and invasion in bovine retinal endothelial cells (BRECs). BRECs were isolated from the retina. Cell viability was confirmed by the MTT assay. In vitro wound migration assay was performed to investigate the migration of BRECs. In vitro tube formation was measured by on-gel system. Apoptosis induced by AuNP was confirmed by caspase-3 assay. AGE-bovine serum albumin (BSA) demonstrated increase of cell migration and proliferation in BRECs. In addition, AuNP regardless of the existence of AGE-BSA suppressed proliferation, migration, and angiogenesis. AuNP suppressed AGE-BSA induced migration and invasion, and induced apoptosis through caspase-3. As a results, AuNP have a potential anti-angiogenic effect for AGE-induced angiogenesis in vitro and offer possibility for the treatment of diabetic retinopathy.

Quantitative Assessment of Xenoestrogenic Environmental Pollutants using E-SCREEN Assay (E-SCREEN Assay를 이용한 내분비계 장애물질의 정량적 평가)

  • 오승민;이상기;정규혁
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.416-423
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    • 2000
  • There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.

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A Minor Transactivation Effect of GATA-3 on its Target Sites in the Extrachromosomal Status

  • Lee, Gap-Ryol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.2056-2060
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    • 2007
  • Transcription factor GATA-3 is the critical transcription factor for Th2 cell differentiation. In spite of its importance in Th2 cell differentiation, the molecular mechanism for its action in Th2 differentiation is poorly understood. Previous studies have suggested that GATA-3 may be involved in the chromatin remodeling in the Th2 cytokine locus. To determine whether GATA-3 exerts its effect on its target sites in the extrachromosomal status, cell transfection assay was performed. In this assay, 800 bp IL4 promoter-luciferase constructs linked with GATA-3 target sites were transfected into the M12 B cell line, D10 mouse Th2 cell lines, and human T lymphoma Jurkat cell lines with or without the GATA-3 expression vector. The GATA-3 effects on its target sites were minimal in the extrachromosomal status, supporting the previous propositions that GATA-3 functions at the chromatin level by remodeling chromatin structure.

Identification of Soil Actinomycetes Producing Anticancer Agent and Its Biological Activities (항암활성물질을 생산하는 토양방선균의 동정 및 함암물질의 생물학적 활성)

  • 박정민;문순옥;오두환
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.347-352
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    • 1994
  • Cytotoxic test was performed by SRB assay on human epidermoid carcinoma HEp-2 cell line for screening the soil microorganism, secreting anticancer agent. One microorganism was selected among two thousand microorganisms for its highest cytotoxicity. And this microorganism was identified with Streptomyces species after performing of diaminopimeric acid and reducing sugar analysis of cell wall and analysing the cultural characteristics and named Streptomyces sp. SM 1119. The effect of anticancer agent in SM 1119 culture extract on the cell cycle was studied by using GG$_{o}$ synchronized NIH 3T3 cells. The extract inhibited the serum stimulation of GG$_{o}$ NIH 3T3 cell only within 1 hour after serum stimulation but not after 6 hours. The extract also reduced the amount of c-myc mRNA in Colo 320 cell. These results suggest that the anticancer agent in the extract inhibits the progression of cell cycle very early stages, probably from G$_{0}$ to G$_{1}$.

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A drug screening system: Preparation of cell islets on the hydrophilic surface

  • Gwon, Hyeok-Seong;Bae, Geun-Won;Jeong, Gyeong-Hui;Kim, Ik-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.581-584
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    • 2002
  • Individual surface(hydrophilic/hydrophobic) were prepared and mammalian cells were cultured on the hydrophilic region. For drug test, cancer and normal cells were treated with Taxol, as an example. Our system was compared with MTT assay. CHO cells were resistant to Taxol up to 100 nM in both Methods. However, A549 cells was sensitive at 100 nM Taxol in the 2 day-treatment. Cervical carcinoma cell, HeLa, was very sensitive to Taxol. In our system, the cells were not shown from above 20 nM Taxol treatment. Our system was competitive to MTT assay in animal cells for drug test.

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The Anthocyanin Components and Cytotoxic Activity of Syzygium cumini (L.) Fruits Growing in Egypt

  • Nazif, Naglaa M.
    • Natural Product Sciences
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    • v.13 no.2
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    • pp.135-139
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    • 2007
  • Four anthocyanins were isolated from the acidic alcoholic extract of Syzygium cumini fruits growing in Egypt: Pelargonidin-3-O-glucoside, pelargonidin-3,5-O-diglucoside, cyanidin-3-O-malonyl glucoside, and delphenidin-3-O-glucoside. They were identified by the chromatographic, TLC and PC, and spectral analyses, UV, $^1$H-NMR and FAB/MS. The fruits were found to contain 0.03 gm % anthocyanins calculated on fresh weight basis calculated by spectrophotometric assay. Cytotoxic activity of total alcoholic extract of the fruits was performed against several types of tumor cell lines using the SRB assay. The tested extract exhibited significant cytotoxic activity for MCF7 (breast carcinoma cell line) (IC$_{50}$= 5.9 ${\mu}$g/mL), while the IC$_{50}$ was > 10 ${\mu}$g/mL for both Hela (Cervix carcinoma cell line), HEPG2 (liver carcinoma cell line), H460 (Lung carcinoma cell line) and U251 (Brain carcinoma cell line).

The Effect of Hyaluronic Acid on the Invasiveness of Malignant Glioma Cells : Comparison of Invasion Potential at Hyaluronic Acid Hydrogel and Matrigel

  • Jin, Shu-Guang;Jeong, Young-Il;Jung, Shin;Ryu, Hyang-Hwa;Jin, Yong-Hao;Kim, In-Young
    • Journal of Korean Neurosurgical Society
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    • v.46 no.5
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    • pp.472-478
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    • 2009
  • Objective : Hyaluronidase (HAse), a degrading enzyme of hyaluronic acid (HA), is highly expressed in patients with malignant glioma. The purpose of this study was to verify whether HAse is related to the invasion of glioma cells. We also investigated if glioma cells with higher mobility in 2-dimensioal (2-D) method have also higher mobility at 3-dimensional (3-D) environment. Methods : Malignant glioma cell lines (U87MG, U251MG, U343MG-A, and U373MG) were used, and their HAse expressions were evaluated by HA zymography. The migration ability was evaluated by simple scratch technique. The invasiveness of each cell lines was evaluated by Matrigel invasion assay and HA hydrogel invasion assay. In HA hydrogel invasion assay, colonies larger than $150\;{\mu}m$ were regarded as positive ones and counted. Statistical analysis of migration ability and invasion properties of each cell lines was performed using t-test. Results : In scratch test to examine migration ability of each cell lines, U87MG cells were most motile than others, and U343MG-A least motile. The HAse was expressed in U251MG and U343MG-A cell lines. However, U87MG and U373MG cell lines did not express HAse activity. In Matrigel invasion assay, the cell lines expressing HAse (U251MG and U343MG-A) were more invasive in the presence of HA than HAse deficient cell lines (U87MG and U373MG). In HA hydrogel invasion assay, the HAse-expressing cell lines formed colonies more invasively than HAse-deficient ones. Conclusion : Malignant Glioma cells expressing HAse were more invasive than HAse-deficient ones in 3-dimensional environment. Therefore, it might be suggested that invasion of malignant gliomas is suppressed by inhibition of HAse expression or HA secretion. Additionally, the ability of 2-D migration and 3-D invasion might not be always coincident to each other in malignant glioma cells.