• 제목/요약/키워드: Micronuclei assay

검색결과 75건 처리시간 0.02초

마우스 골수세포의 중기염색체 분석 및 미소핵 검사를 이용한 피폭선량 평가법의 개발 (Development of a Noble Dosimetry Using Metaphase Analysis and Micronuclei Assay of Bone Marrow Cells in Mice)

  • 민정준;범희승;김영호;윤현중;김지열
    • 대한핵의학회지
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    • 제34권1호
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    • pp.74-81
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    • 2000
  • 목적: 마우스를 전신조사한 다음 골수세포를 추출하여 중기염색체 분석법과 미소핵 검사법으로 방사선량별 염색체 이상의 빈도를 관찰하고 이를 통하여 표준선량반응곡선을 얻어내어 방사선이 인체 골수세포에 미치는 영향을 추정하기 위해 이 연구를 시행하였다. 대상 및 방법: ICR계 마우스를 대상으로 각각 0, 1, 2, 3, 4, 5, 10 Gy를 조사하고 대퇴골의 골수를 추출하여 중기염색체 분석법을 시행하였고, 0, 1, 2, 3, 4, Gy를 조사한 후 미소핵 분석법을 시행하였다. 각각의 조사량에 따라 중기염색체에서 이중 중심체형 염색체와 반지형 염색체를 계수하였고, 다염성적혈구에서 관찰된 미소핵을 계수하였다. 결과: 중기염색체에서 이중 중심체형 염색체와 반지형 염색체의 빈도를 나타내는 Ydr 값은 0, 1, 2, 3, 4, 5, 10 Gy에서 각각 0.002, 0.107, 0.33, 0.625, 1.055, 1.662, 5.843이었고 선량-반응관계를 선형회귀분석한 결과 방사선량(D)과 염색체이상 빈도(YDR)와의 관계는 YDR=0.0176+0.0324D+0.0567$D^2$ (r=1.0, p<0.001)으로 나타났다. 다염성적혈구에서 관찰된 미소핵은 0, 1, 2, 3, 4, Gy에서 각각 0.001, 0.062, 0.218, 0.478, 0.841로 방사선량(D)과 미소핵의 빈도(mn)와의 관계는 F(mn)=0.0019+0.0073D+0.00506$D^2$ (r=1.0, p<0.001)로 나타나 선량에 따른 염색체이상의 빈도는 이차함수식으로 증가함을 알 수 있었다. 두 가지의 검사방법간에는 매우 강한 상관관계를 나타내었다(r=0.99, p<0.01). 결과: 마우스의 골수세포에서 중기염색체 분석법과 미소핵 검사법은 생체 내의 피폭선량을 평가하는데 매우 유용하였고, 이 두 가지 검사법 중 어느 방법을 사용하여도 방사선에 의한 생물학적 효과를 평가할 수 있을 것으로 사료되었다.

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생쥐 골수세포에서 아드리아마이신의 소핵생성에 미치는 N-마세틸시스테인의 억제효과 (Suppressive Effect of N-Acetylcysteine on the Adriamycin-Induced Micronuclei Formation in Mouse Bone-marrow Cells)

  • 손수정;허인회;최성규;허문영
    • 약학회지
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    • 제37권3호
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    • pp.278-285
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    • 1993
  • The anticlastogenic effect of N-acetylcysteine was tested in vivo in mouse bone-marrow micronucleus assay. The frequencies of micronuclei induced by adriamycin (5 mg/kg i.p.) in bonemarrow cells were decreased by the oral administration of N-acetylcysteine at 12 h before adriamycin injection. The observed suppressing effect was not a reflection of a delay in the formation of micronuclei by the cytotoxic effect of N-acetylcysteine. The anticlastogenic effects of SH compound including N-acetylcysteine, cysteine, cystine, S-carboxy methylcysteine and glutathione were also investigated by the multiple pretreatment. Each SH compound was administered orally every day for 5 days and adriamycin (5 mg/kg i.p.) was injected at 24h after the last dose of test compound. N-acetylcysteine and glutathione showed significantly the suppressive effect at dose of 10 and 25 mg/kg for N-acetylcysteine and at the dose of 25 mg/kg for glutathione. Our study suggests that N-acetylcysteine is capable of protecting the chromosomal damages in the normal cells during cancer chemotherapy by adriamycin, and may act as an anticlastogen against induction of micronuclei by superoxide generating agent such as adriamycin.

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

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • 한국환경성돌연변이발암원학회지
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    • 제21권1호
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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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Radiation exposure dose in human blood lymphocytes as assessed by the CBMN assay

  • Ryu, Tae Ho;Kim, Jin-Hong;Kim, Jin Kyu
    • Journal of Ecology and Environment
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    • 제37권4호
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    • pp.195-200
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    • 2014
  • The chances of accidental exposure are augmented as the application of ionizing radiation increases in various fields. Such accidental exposures may occur at nuclear power plants, laboratories, and hospitals. Cytogenetic assays have been used for estimating radiation dose in the situation of the accidents. The micronucleus assay has several advantages over the other cytogenetic methods as it is simple and fast. The present study aimed at investigation of the micronuclei frequencies in cytokinesis-block cells in human blood lymphocytes after ${\gamma}$-irradiation and at establishment of a standard dose response relationship. The samples of peripheral blood were obtained from 6 different donors aged between 24 and 30 years old. The bloods were irradiated in vitro with 0-5 Gy. A linear quadratic dose-response equation was obtained by scoring the micronuclei in binucleated cells; $y=27.87x^2+46.13x+2.08$ ($r^2=0.99$). Irradiation caused a significant decrease in the nuclear division index. Necrotic and apoptotic cells increased in number after irradiation in a dose-dependent manner. In conclusion, the conventional cytokinesis-block micronucleus assay has proven to be the great technique in biological dosimetry. Dose-response calibration curve derived from CMBN assay could be used to estimate the exposure dose during a radiological emergency.

Protection against Whole Body γ-Irradiation Induced Oxidative Stress and Clastogenic Damage in Mice by Ginger Essential Oil

  • Jeena, Kottarapat;Liju, Vijayasteltar B;Ramanath, Viswanathan;Kuttan, Ramadasan
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권3호
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    • pp.1325-1332
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    • 2016
  • Radioprotective effects of ginger essential oil (GEO) on mortality, body weight alteration, hematological parameters, antioxidant status and chromosomal damage were studied in irradiated mice. Regression analysis of survival data in mice exposed to radiation yielded LD50/30 as 7.12 and 10.14 Gy for control (irradiation alone) and experimental (GEO-treated irradiated) mice, respectively, with a dose reduction factor (DRF) of 1.42. In mice exposed to whole-body gamma-irradiation (6 Gy), GEO pre-treatment at 100 and 500 mg/kg b.wt (orally) significantly ameliorated decreased hematological and immunological parameters. Radiation induced reduction in intestinal tissue antioxidant enzyme levels such as superoxide dismutase, catalase, glutathione peroxidase and glutathione was also reversed following administration of GEO. Tissue architecture of small intestine which was damaged following irradiation was improved upon administration of GEO. Anticlastogenic effects of GEO were studied by micronuclei assay, chromosomal aberration and alkaline gel electrophoresis assay. GEO significantly decreased the formation of micronuclei, increased the P/N ratio, inhibited the formation of chromosomal aberrations and protected agaisnt cellular DNA damage in bone marrow cells as revealed by comet assay. These results are supportive of use of GEO as a potential radioprotective compound.

Argyrophilic nucleolar organizer region(AgNOR) 염색을 이용한 림프구의 미세핵분석법의 개선 (Improvement of micronucleus assay in the lymphocytes using Argyrophilic nucleolor organizer region(AgNOR) staining)

  • 김성호;한동운;강문일;임정택
    • 대한수의학회지
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    • 제36권2호
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    • pp.277-281
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    • 1996
  • A technique to improve the analysis of micronuclei(MN) in lymphocytes as a cytogenetic indicator is reported. For the purpose of diminishing the variation of the result from individual reader and making it easier to distinguish accurately a cytokinesis blicked(CB) lymphocyte and micronuclei, we tried a modified one-step silver staining technique as a method for detection of the argyrophilic nucleolar organizer region(AgNOR) with or without conventional Giemsa stain in the slide from CB method. Compared with the conventional Giemsa stain, the preparation processed with this method are especially useful for the accurate analysis of MN of cultured lymphocyte with cytochalasin B. This method will be a useful technique for automated calculation of MN.

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림프구의 미소핵을 지표로 영광 원자력발전소 주변 사육 돼지의 방사선 생물학적 평가 (Radiobiological Evaluation in Pig Bred in the Vicinity of Yeonggwang Nuclear Power Station Using Micronuclei in Cytokinesis-blocked Lymphocyte)

  • 김세라;강창모;김성호
    • 한국임상수의학회지
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    • 제21권3호
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    • pp.286-290
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    • 2004
  • Cytogenetic and hematological analysis was performed in peripheral blood of pig in the vicinity of Yeonggwang nuclear power station and control area. The frequency of micronuclei (MN) in peripheral blood lymphocytes from pig was used as a biomarker of radiobiological effects resulting from exposure to environmental radiation. An estimated dose of radiation was calculated by a best fitting linear-quadratic model based on the radiation-induced MN formation from the swine lymphocytes exposed in vitro to radiation over the range from 0 Gy to 4 Gy. MN rates in lymphocytes of pig from Yeonggwang nuclear power station and control area were 10.60/1,000 and 11.10/1,000, respectively. There were no significant differences in MN frequencies and hematological values in pig between Yeonggwang and control area. The study indicates that the MN assay in lymphocyte of pig is a rapid, sensitive and accurate method that can be used to monitor a large population exposed to radiation.

PROTECTION OF DNA BY SCUTELLARlA BAICALENSIS IN HL-60 CELLS EXPOSED TO $\gamma$-RAYS; ANALYSED BY MICRONUCLEI FORMATION AND SINGLE CELL GELL ELECTROPHORESIS

  • Heon Oh;Park, Hae-Ran;Ham, Yeon-Ho;Kim, Sung-Ho;Jo, Sung-Kee
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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    • pp.107-107
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    • 2001
  • In the present study, the protective effect of Scutellaria baicalensis against DNA damage in HL-60 cells exposed to $^{60}$ Co ${\gamma}$-rays was evaluated using micronuclei formation and alkaline single cell gel electrophoresis (SCGE, comet assay). The frequency of micronuclei was decreased in groups treated with water extract (P<0.01), polysaccaride fraction (P<0.01) and methanol fraction (P<0.01) before/after exposure to 200 cGy of ${\gamma}$-rays.(omitted)

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에틸렌옥사이드(Ethylene oxide)에 노출된 병원 근로자들의 소핵 빈도와 유전적 감수성 지표와의 연관성 (Analysis of Micronuclei and Its Association with Genetic Polymorphisms in Hospital Workers Exposed to Ethylene Oxide)

  • 이선영;김양지;최영주;이중원;이영현;신미연;김원;윤충식;김성균;정해원
    • 한국환경보건학회지
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    • 제37권6호
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    • pp.429-439
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    • 2011
  • Objectives: Ethylene oxide (EtO) is classified as a human carcinogen, but EtO is still widely used to sterilize heat-sensitive materials in hospitals. Employees working around sterilizers are exposed to EtO after sterilization. The aim of the present study was to assess the exposure of EtO level, coupled with occupationally induced micronuclei from hospital workers. The influence of genetic polymorphisms of detoxifying genes (GSTT1 and GSTM1) and DNA repair genes (XRCC1 and XRCC3) on the frequencies of micronuclei in relation to exposure of EtO was also investigated. Methods: The study population was composed of 35 occupationally exposed workers to EtO, 18 student controls and 44 unexposed hospital controls in Korea. Exposure to EtO is measured by passive personal samplers. We analyzed the frequencies of micronuclei by performing cytokinesis-block micronucleus assay (CBMN assay) and GSTM1, GSTT1, XRCC1, and XRCC3 were also genotyped by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequencies of micronuclei in EtO exposure group, student controls and hospital controls were $18.00{\pm}7.73$, $10.47{\pm}7.96$ and $13.86{\pm}6.35$ respectively and their differences were statistically significant, but no significant differences according to the level of EtO were observed. There was a dose-response relationship between the frequencies of micronuclei and cumulative dose of EtO, but no significantly differences were observed. We also investigated the influence of genetic polymorphisms (GSTM1, GSTT1, XRCC1, and XRCC3) on the frequencies of micronuclei, but there were no differences in the frequencies of micronuclei by genetic polymorphisms. Conclusions: The frequencies of micronuclei in EtO exposure group was significantly higher than control groups. A dose-response relationship was found between the level of EtO exposure and the frequencies of micronuclei, but no statistically differences were observed. We also found that the frequencies of micronuclei were increased according to cumulative EtO level. There was no association of the genetic GSTM1, GSTT1, XRCC1, and XRCC3 state with the frequency of micronuclei induced by EtO exposure.

Adaptive Response Induced by Low Dose Ionizing Raditation in Human Cervical Carcinoma Cells

  • Kim, Jeong -Hee;Lee, Kyung -Jong;Cho, Chul -Koo;Yoo, Seong -Yul;Kim, Tae -Hwan;Ji, Young -Hoon;Kim, Sung -Ho
    • Archives of Pharmacal Research
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    • 제18권6호
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    • pp.410-414
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    • 1995
  • Adaptive response induced by low dese .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray (1-cGy) followed by high doses of r-ray irradiation (0,1,2,3,5,7 and 9Gy for chlnogenic assay or 1.5Gy for micronucleus assay) with various time intervals. Survival fractions of cells in both low dose-irradiated and unirrated groups were analyzed by clonogenic assay. Surviva fractions of low dose-irradiated in cell survival was maximum when low and high dose irradiation time interval was 4 hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enutained from survival fractions analyzed by clonogenic assay, maximum when low and high dose irradiation time interval was 4hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enumerated in both low dose-irradiated and unirradiated groups. In consiststent with the result obtained from survival fractions analyzed by clonogenic assay, maximum reduction in frquencies of micronuclei was observed when low dose radiation was given 4 hr prior to high response to subsequent high dose .gamma.-ray irradiation in human cervical carcinomal cells. Our data suggest that one of the possible mechanisms of adaptive response induced by low dose rediation is the increase in repair of DNA double strand breaks in low dose radiation-adapted cells.

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