• 한국환경성돌연변이발암원학회지
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• 제17권2호
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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.

• 한국환경생물학회:학술대회논문집
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• 한국환경생물학회 2003년도 학술대회
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• pp.68-73
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• 2003

The possibility of using the alkaline protocol of the single cell gel electrophoresis (SCGE) assay as a method for detecting induced DNA damage has been studied for six major plants. The EMS was applied as a model genotoxic agent on young excised leaves of the tested crops for 18 h at 26$^{\circ}C$ in the dark. With increasing concentrations of 0 to 10 mM EMS, the DNA damage, expressed by the averaged median tail moment values, significantly increased in nuclei of all plants studied. As the results, no correlation between the diameter of nuclei and sensitivity to EMS treatment was observed. The data obtained demonstrate the feasibility of using the SCGE assay for detecting induced DNA damage in plants.

• Toxicological Research
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• 제20권3호
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• pp.225-232
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• 2004

The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01～1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.176-176
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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 assayed.(omitted)

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2004년도 춘계학술대회
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• pp.95-95
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• 2004

• Toxicological Research
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• 제23권1호
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• pp.25-31
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• 2007

To evaluate the protective effect of Houttuynia cordata on hydrogen peroxide-induced oxidative DNA damage in HepG2 cell line, we used an alkaline single-cell gel electrophoresis (SCGE; comet assay). The DNA damage was analyzed by tail moment (TM) and tail length (TL), which used markers of DNA strand breaks in SCGE. The $100{\mu}g/ml$ of methanolic extract of Houttuynia cordata root showed significant protective effects (p < 0.01) against hydrogen peroxide-induced DNA damage in HepG2 cells and increased cell viability against hydrogen peroxide. The results of this study indicate that Houttuynia cordata root methanol extract acts as a potential antioxidant, and exhibits potential anticancer properties, which may provide a clue to find applications in new pharmaceuticals for oxidative stability.

• 한국식품위생안전성학회지
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• 제12권4호
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• pp.315-320
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• 1997

Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. This study was devised to develop a chemopreventive agent from panax ginseng to be able to suppress the genotoxicity and oxidative damage by ractive oxygen species, which are involved with cancer or aging. Ginseng petroleum ether extract (GPE) and one of its fraction, P2, showed an antioxidative effect on the lipid peroxidiphenyl-2-picryl hydrazil (DppH) radical generation. They also showed the suppressive effect of H2O2 or KO2 induced DNA damage by single cell gel electrophoresis (SCGE). Results from our study indicate that GPE and P2 are capable of protecting lipid peroxidation, and oxidative DNA damage. Therefore, GPE and P2 may be useful chempreventive agents which are involved with cancer and aging.

• 한국환경성돌연변이발암원학회지
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• 제21권2호
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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.

• 대한수의학회지
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• 제44권1호
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• pp.41-47
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• 2004

The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to detect DNA damage induced by a number of chemicals and biological factors in vivo and in vitro. The DNA damage was analysed by tail moment (TM) and tail length (TL), which were markers of DNA strand breaks in SCGE. Human, mouse and rat peripheral blood lymphocytes (PBLs) were irradiated with different doses of $^{60}Co$ ${\gamma}$-rays, e.g. 1, 2, 4, and 8 Gy at a dose rate of 1 Gy/min. A dose-dependent increase in TM (p<0.01) and TL (p<0.01) was obtained at all the radiation doses (1-8 Gy) in human, mouse and rat PBLs. Mouse PBLs were more sensitive than human PBLs which were in turn more sensitive than rat PBLs when the treated dosages were 1 and 2 Gy. However, human PBLs were more sensitive than mouse PBLs which were in turn more sensitive than rat PBLs when the irradiation dosages were 4 and 8 Gy. Data from all three species could be fitted to a linear-quadratic model. These results indicated that there may be inherent differences in the radio-sensitivity among PBLs of mammalian species.

• Journal of Radiation Protection and Research
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• 제24권2호
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• pp.93-99
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• 1999

Alkaline single cell gel electrophoresis (SCGE) assay는 일명 혜성분석이라고 부르며 in vivo 와 in vitro 에서 많은 화학적, 생물학적인 인자에 의한 DNA 손상을 감지하는데 유용한 기법으로 각각의 세포에서 DNA 단일 가닥 절단과 알칼리에 약한 장소를 평가하는 새로운 방법으로 인정되고 있다. 단세포 겔 전기영동법 (SCGE)을 사용하여 복숭아씨 추출물이 방사선에 의하여 사람 림프구 DNA에 나타나는 손상을 보호하는 지 여부를 평가하였다. 복숭아씨 추출물로 10 분간 전처리한 림프구를 0, 0.1, 0.3, 0.5, 1.0, 2.0 Gy 의 방사선으로 조사하였고 방사선만을 조사한 림프구 실험군과 비교평가하였다. 혜성분석에서 DNA 가닥 절단에 대한 표식인 tail moment의 증가는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었으며 각각의 농도별로 복숭아씨 추출물이 처리된 림프구의 DNA 손상은 현저히 감소하였다. 단세포 겔 전기영동법을 통한 평가결과 복숭아씨 추출물은 방사선에 의한 림프구 DNA 손상에 대한 탁월한 방어효과를 나타내었다.