• 제목/요약/키워드: DNA single-strand breaks

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김치로부터 분리된 우세 균주들의 in vitro 항유전 독성효과 (Antigenotoxic Effect of Dominant Bacterial Isolates from Kimchi in vitro)

  • 최준원;박종흠;지승택;최옥병;신현길
    • 한국식품과학회지
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    • 제31권4호
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    • pp.1071-1076
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    • 1999
  • 본 연구에서는 김치발효에 관여하는 우세 발효균들의 집락을 분리하고, 발암원인 MNNG에 대하여 이들 분리주들이 갖는 DNA 손상 억제효과를 조사하였다. 이를 위해서 세포의 DNA 손상 및 복구를 측정하는데 이용되는 매우 신속하고 감도가 높은 Single Cell Gel Electrophoresis(SCGE 법, 또는 comet assay)를 사용하였다. 배추김치에서 분리한 3개의 분리주중에서 B-3를 제외한 B-1과 B-2는 양성 대조구와 비교하였을 때 우수한 항유전 독성효과를 나타내었으며(p<0.01), 율무김치의 3 개의 분리주(Y-1, Y-2, Y-3)와 1개의 분리주(Y-4)도 각각 항유전 독성효과를 나타내었다(p<0.05, p<0.01). 총각김치의 5 개 분리주중에서 3개의 분리주와 깍두기의 9개의 분리주중에서 2 개의 분리주들도 각각 항유전 독성효과를 나타내었다(p<0.05, p<0.01).

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Genotoxicity and Identification of Differentially Expressed Genes of Formaldehyde in human Jurkat Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제1권4호
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    • pp.230-236
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    • 2005
  • Formaldehyde is a common environmental contaminant found in tobacco smoke, paint, garments, diesel and exhaust, and medical and industrial products. Formaldehyde has been considered to be potentially carcinogenic, making it a subject of major environmental concern. However, only a little information on the mechanism of immunological sensitization and asthma by this compound has been known. So, we performed with Jurkat cell line, a human T lymphocyte, to assess the induction of DNA damage and to identify the DEGs related to immune response or toxicity by formaldehyde. In this study, we investigated the induction of DNA single strand breaks by formaldehyde using single cell gel electrophoresis assay (comet assay). And we compared gene expression between control and formaldehyde treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity ($IC_{30}$) of formaldehyde was determined above the 0.65 mM in Jurkat cell in 48 h treatment. Based on the $IC_{30}$ value from cytotoxicity test, we performed the comet assay in this concentration. From these results, 0.65 mM of formaldehyde was not revealed significant DNA damages in the absence of S-9 metabolic activation system. And the one differentially expressed gene (DEG) of formaldehyde was identified to zinc finger protein 292 using $GeneFishing^{TM}$ method. Through further investigation, we will identify more meaningful and useful DEGs on formaldehyde, and then can get the information on the associated mechanism and pathway with immune response or other toxicity by formaldehyde exposure.

Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

  • Kang, Changgeun;Lee, Hyungkyoung;Yoo, Yong-San;Hah, Do-Yun;Kim, Chung Hui;Kim, Euikyung;Kim, Jong Shu
    • Toxicological Research
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    • 제29권1호
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    • pp.43-52
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    • 2013
  • Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 ${\mu}M$) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 ${\mu}M$). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.

홍삼 추출물에 의한 유전독성 감소효과 (I) - 배양 NIH3T3 세포에서 자외선에 의한 유전독성의 감소에 미치는 홍삼추출물 처리효과 (Decrease of Genotoxicity by Red Ginseng Root Extract (I) - Decrease of UV -induced Genotoxicity by Red Ginseng Root Extract in Cultured NIH3T3 Cells)

  • 김완주;유병수
    • 대한화장품학회지
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    • 제24권1호
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    • pp.74-86
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    • 1998
  • 자외선에 의한 유전독성의 감소에 미치는 홍삼추출물의 영향을 배양 NIH3T3 세포계에서 분석하였다. 자외선을 조사한 후 정상 배지에서 배양한 시간간격에 따라 세포의 생존률은 증가하였는데 홍삼추출물이 함유된 배지에서 배양한 경우는 약 15%정도 증가한 생존률을 보였다. 자외선을 조사한 후 감소된 DNA복제가 정상배지 배양시간에 따라 증가하는 정도도 홍삼추출물을 후처리할 경우 현저한 증가를 보였다. 자외선 상해를 회복하기 위한 절제회복능은 홍삼추출물을 처리할 경우 유의미한 증가를 보였다. 이러한 절제회복과정 중 효소에 의한 절제단계가 홍삼추출물 처리에 의해 활성화됨을 단사절단 분석을 통하여 규명하였다. 이상의 결과는 홍삼추출물이 자외선 상해의 절제회복에 유의미한 증가를 보이며 따라서 유전독성을 감소시키는 항노화제로써 사용할 수 있음을 시사한다.

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T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성 (Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment)

  • 하대식;허정호;이국천;조명희;김국헌;김충희;류재두;이승환;김곤섭;김의경;김종수
    • Toxicological Research
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    • 제22권2호
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Jung, Sang-Hun;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 춘계학술대회
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    • pp.185-185
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase (tk$\^$+/-/) gene assay (MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. In MOLY assay, JSH-Ⅶ-3 at 50 ∼ 6 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in the absence and presence of S-9 metabolic activation system. However, the concentration of ISH-II-3, 38 $\mu\textrm{g}$/ml, induced increased mutation frequency (MF) in the presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in JSH-Ⅵ-3 and JSH-Ⅶ-3, wherase concentration of 32.8 $\mu\textrm{g}$/ml in JSH-II-3 and 13.9 $\mu\textrm{g}$/ml in JSH-Ⅶ-3 were induced DNA damage in the absence of S-9 metabolic activation system. Therefore, we suggest that JSH-Ⅵ-3 and JSH-Ⅶ-3 have no genotoxic effects but JSH-II-3 and JSH-Ⅷ-3 induce some mutagenicity and DNA strand breaks in mouse lymphoma cell line used this study.

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Selective Effects of Curcumin on CdSe/ZnS Quantum-dot-induced Phototoxicity Using UVA Irradiation in Normal Human Lymphocytes and Leukemia Cells

  • Goo, Soomin;Choi, Young Joo;Lee, Younghyun;Lee, Sunyeong;Chung, Hai Won
    • Toxicological Research
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    • 제29권1호
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    • pp.35-42
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    • 2013
  • Quantum dots (QDs) have received considerable attention due to their potential role in photosensitization during photodynamic therapy. Although QDS are attractive nanomaterials due to their novel and unique physicochemical properties, concerns about their toxicity remain. We suggest a combination strategy, CdSe/ZnS QDs together with curcumin, a natural yellow pigment from turmeric, to reduce QD-induced cytotoxicity. The aim of this study was to explore a potentially effective cancer treatment: co-exposure of HL-60 cells and human normal lymphocytes to CdSe/ZnS QDs and curcumin. Cell viability, apoptosis, reactive oxygen species (ROS) generation, and DNA damage induced by QDs and/or curcumin with or without ultraviolet A (UVA) irradiation were evaluated in both HL-60 cells and normal lymphocytes. In HL-60 cells, cell death, apoptosis, ROS generation, and single/double DNA strand breaks induced by QDs were enhanced by treatment with curcumin and UVA irradiation. The protective effects of curcumin on cell viability, apoptosis, and ROS generation were observed in normal lymphocytes, but not leukemia cells. These results demonstrated that treatment with QD combined with curcumin increased cell death in HL-60 cells, which was mediated by ROS generation. However, curcumin acted as an antioxidant in cultured human normal lymphocytes.

이온화 방사선 및 염화수은(II)에 의한 자궁경부암 세포의 DNA 손상 평가 (Evaluation of DNA Damage by Mercury Chloride (II) and Ionizing Radiation in HeLa Cells)

  • 우현정;김지향;안토니나 체불스카바실레프스카;김진규
    • 환경생물
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    • 제24권1호
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    • pp.46-52
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    • 2006
  • 세포에 미치는 염화수은(II)과 이온화 방사선의 영향과 수은 처리 전 후 방사선 조사 시 그 상호 작용에 관해 알아보고자 본 연구를 수행하였다. 염화수은(II)의 독성정도를 알아보기 위하여 사람의 자궁암 세포에 농도별로 염화수은(II)을 처리하였다. 세포의 생존율은 3가지 농도(1,0. 1,0. $0.01\;{\mu}M$)모두에서 유의하게 감소하였으며 이미 $0.1\;{\mu}M$에서 약 73%의 생존율이 감소하는 것으로 나타났다. 염화수은(II)과 방사선의 단독처리 시 DNA의 손상 정도에 비해 복합처리 시의 DNA손상 정도가 $2\sim4$배 정도 확연히 높아짐을 볼 수 있었다. 특히 방사선 후 수은 처리군은 DNA손상의 정도가 다른 처리군에 비하여 높게 나타났는데 이는 이미 기존의 보문에서 밝혀진 바와 같이 수은의 DNA수복에 관련되어 있는 Fpg protein에 미치는 영향 때문으로 사료된다. 이미 방사선에 의해 산화적 손상을 입은 DNA의 수복 기작을 수은이 방해하여 좀 더 높은 손상을 가져오는 것을 확인할 수 있었다.

Responses of Various Biomarkers in Common Carp (Cyprinus carpio) Exposed to Benzo[k]fluoranthene

  • Kim, Woo-Keun;Kim, Ja-Hyun;Yeom, Dong-Hyuk;Lee, Sung-Kyu
    • 생태와환경
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    • 제41권3호
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    • pp.331-337
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    • 2008
  • Polycyclic aromatic hydrocarbons (PAHs) derived from leakage of fossil fuels and incomplete combustion of organic materials have been considered as harmful contaminants in environments. This study evaluated the effect of benzo[k]fluoranthene (BkF), one of the PAHs, using the multiple biomarkers and applied the integration model with those biomarker responses. After 10 days of the exposure at the measured concentrations of BkF (6, 25, and 45 ${\mu}g\;L^{-1}$), the changes of the four biomarkers, that is, 7-ethoxyresorufin-O-deethylase (EROD), DNA single-strand breaks (Comet), acetylcholinesterase (AChE) and vitellogenin (VTG) in the common carp (Cyprinus carpio) were observed. The standardized values of four biomarker responses were computed and integrated as star plots, representing Integrated Biomarker Respnse (IBR) values. DNA damage was induced in a dose-dependent manner, and increased significantly compared with that in the control. EROD and VTG levels were significantly elevated at low concentrations of BkF. On the other hand, AChE activities were not altered by BkF. IBR values increased as the exposure concentrations increased. Thus, the metabolic, endocrine and genetic changes of the biomarker responses in the common carp exposed to BkF should be considered in the case of the ecological risk assessment of the BkF in fish and it can be used as a biomonitoring tool in aquatic ecosystems. In addition, star plots can be used as a useful analysis tool in multibiomarker integration approach.