• Title/Summary/Keyword: genotoxicity test

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Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level (DNA damage와 Apoptosis를 정량화하는 단세포전기영동법)

  • 류재천;김현주;서영록;김경란
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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Genotoxicily Studies of An Anticancer Agent of Camptothecin Series, CKD-602 (Camptothecin계 항암제 CKD-602의 유전독성평가)

  • 하광원;오혜영;허옥순;박장환;손수정;한의식;김종원;강일현;강혁준
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.129-134
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    • 1998
  • To evaluate the genotoxicity of CKD-602, an anticancer agent the in viかo reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the reverse mutation assay, CKD-602 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, and TA 1537 strains with and without metabolic activation. In the chromosome aberration test using CHL cells, there was an increased incidence of structural aberrations induced by CKD-602 without metabolic activation during 24 and 48 hours, but CKD-602 did not induce chromosome aberration with metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice. At 24 hours after treatment with CED-602 by i.p. once, there was an increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice.

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Genotoxicify Studies of on Antihypertensive Agent, SKP-450 (고혈압 치료제 SKP-450의 유전독성평가)

  • 하광원;오혜영;박장환;허옥순;손수정;한의식;류근호;조용백
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.123-128
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    • 1998
  • To evaluate the genotoxicity of SKP-450, an antihypertensive agent the in vitro reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the Reverse mutation test, SKP-450 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 with and without metabolic activation. In the chromosome aberration assay using CHL cells, there was no increased incidence of structural and numerical aberrations with and without metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice at 30 hours after treatment with SKP-450 by p.o once. The results showed no increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice treated with SKP-450.

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Analysis of chromosome aberration, sister chromatid exchange, micronuclei and single cell gel electrophoresis in human lymphocytes exposed in vitro to Bisphenol A and Diethylstilbestrol (비스페놀 A 및 Diethylstilbestrol의 유전독성 평가를 위한 염색체이상, 자매염색분체교환, 소핵형성, 단일세포 겔 전기영동법의 활용)

  • 김병모;정해원
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.135-141
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    • 2001
  • Endocrine disruptors have been implicated in carcinogenesis in animal studies, but carcinogenetic effects on human remain controversial. In order to examine the genotoxicity of two common endocrine disruptors, Bisphenol A and Diethylstilbestrol, cytogenetic endpoints including chromosome aberration (CA), sister chromatid exchange (SCE), micronuclei (MN) analyses and DNA damage by single cell gel electrophoresis (SCGE) were assessed. The effects of Bisphenol A and Diethylstilbestrol on the frequencies of CA and MN were increased in a dose-dependent manner and that of Bispheol A was more significant by Kendall'$\tau$test. Bisphenol A and Diethylstilbestrol also increased the frequency of SCE. Bisphenol A and Diethylstilbestrol induced DNA damage in a dose-dependent manner and the DNA damage induced by Diethylstilbestrol in human blood lymphocytes was more significant.

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In vitro and In vivo Evaluation of Genotoxicity of Stevioside and Steviol, Natural Sweetner (천연감미료 스테비오사이드와 스테비올의 생체내, 시험관내 유전독성평가)

  • 오혜영;한의식;최돈웅;김종원;손수정;엄미옥;강일현;강혁준;하광원
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.614-622
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    • 1999
  • The standard operation procedure of mouse lymphoma L5178Y $tk^{+/-}-3.7.2C$ gene mutation assay (MOLY) has been regarded as a sensitive in vitro mammalian cell gene mutation assay that is capable of detecting clastogens as well as mutagens. Using MOLY, one of natural sweetner, stevioside (5mg/ml) and its aglycon, steviol ($340{\;}\mu\textrm{g}/ml$) were evaluated the mutagenicity. Stevioside and steviol did not induce mutagenicity in MOLY. On the other hand, stevioside (250mg/kg, B.W.) and steviol (200mg/kg, B.W.) were also evaluated their ability to induce micronuclei in regenerating hepatocytes and bone marrow cells of ddY mice. From these results, stevioside and steviol did not induce any mutagenic effect both MOLY and in vivo micronucleus test.

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Genotoxic evaluation of Gyllus bimaculatus in 3 sets of mutagenicity tests

  • Ahn, Mi-Young;Pae, Hey-Jin;Lee, Byung-Mu;Ryu, Kang-Sun
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.120.1-120.1
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    • 2003
  • The mutagenic potential of Gryllus bimaculatus was evaluated using the short-term genotoxicity tests including Ames, chromosome aberration and micronuclei tests. In salmonella typhimurium assay, G. bimaculatus did not show any mutagenic response in the absence or presence of S9 mix with TA98, TAl00, TA1535, and TA1537. In chromosome aberration test, G. bimaculatus did not show any significant effect on Chinese Hamster Ovary (CHO) cells compared with control. (omitted)

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In vitro mutagenicity and genotoxicity study of PAHs and nitro-PAHs using the bacterial revertant (Ames) test and alkaline single cell gel electrophorosis (Comet) assay

  • Kim, Soung-Ho;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.284.1-284.1
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    • 2002
  • In previous studies, we demonstrated that ambient PM collected from urban site of Korea air could induce DNA damage, Various mutagens and carcinogens present in the urban air differ according to the source of the pollutants. Polycyclic aromatic hydrocarbons (PAHs) and their nitrated compound are produced in the combustion of fossil fuels as diesel emission exhausts. In recent, PAH and nitro-PAH have been identified in urban air particulate matter (PM). and some of them were found to be tumorigenic in experimental animals and humans. (omitted)

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Antigenotoxicity of Galangin and its Action Mechanism (Galangin의 유전독성 억제효과와 작용기전)

  • 허문영;류재천
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.77-82
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    • 1998
  • In order to compare the suppressive effect of galangin on the genotoxicity by N-methyl-N-nitrosourea (MNU) or benzo[a]pyrene B(a)P, in vivo micronycleus test using mouse peripheral blood and in vitro sister chromatid exchange(SCE) test using mouse spleen lymphocytes were performed. MNU or B(a)P-induced micronucleated reticulocytes in vivo was decreased by the simultaneous treatment of galangin. MNU or B(a)P-induced SCEs in vitro was also decreased by the simultaneous treatment of galangin. On the other hand, the determinations of [$^3$H]MNU-induced total DNA binding and methylated DNA were performed to find out the mechanism of action. [$^3$H]MNU-induced total DNA binding was inhibited by the treatment of galangin in calf thymus DNA. HPLC analysis of DNA hydrolysates showed that galangin caused a decrease of 7-methyl guanine and $O^{6}$-methyl guanine in calf thymus DNA. To elucidate the action mechanism of galangin against B(a)P, alteration of B(a)P metabolism was studied. Galangin inhibited B(a)P metabolism in the presence of S-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with S-9 mix.

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DNA Damage and Micronuclei Induced by Di (2-ethylhexyl) phthalate in Human Breast Carcinoma MCF-7 cells (Di(2-ethylhexyl) phthalate에 의해 유도된 DNA손상과 소핵 형성)

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • Environmental Mutagens and Carcinogens
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    • v.21 no.1
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    • pp.34-43
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    • 2001
  • Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

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