• Title, Summary, Keyword: Cytokinesis-block micronucleus assay

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The induction of Micronucleus and Aneuploidy in human lymphocytes by Hydroquinone and its association with Genetic Polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene

  • Chung, Eun-Jung;Kim, Tae-Yon;Kim, Jin-Sik;Kim, Yang-Ji;Cho, Yoon-Hee;Chung, Hai-Won
    • Proceedings of the Korean Society of Toxicology Conference
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    • pp.176-176
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    • 2002
  • In order to investigate whether the induction of micronucleus and aneuploidy in human lymphocytes by Hydroquinone (HQ) is associated with genetic polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene, the cytokinesis-block micronucleus (CBMN) assay in combination with fluorescence in situ hybridization (FISH) technique using specific centromeric probes for chromosome 7 and 8 and PCR-RFLP based genotyping for 30 healthy people were performed.(omitted)

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THE COMBINATION OF MICRONUCLEUS ASSAY AND FISH TECHNIQUE FOR THE EVALUATION OF GENOTOXICITY OF 1, 2, 4-BENZENETRIOL

  • Chung, Hai-Won;Kang, Su-Jin;Kim, Su-Young
    • Proceedings of the Korean Society of Toxicology Conference
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    • pp.111-111
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    • 2001
  • The cytokinesis-block micronucleus assay have been emerged as one of the preferred method for assessing chromosome damage. Micronucleus are small, extranuclear bodies that are formed in mitosis from acentric chromosomal fragments or chromosome that are not included in each daughter nuclei.(omitted)

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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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    • v.37 no.4
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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.

Protective Effect of Buplueri Radix (BR) Against 1,2,4-benzentriol Induced DNA Damage in Human Lymphocytes (Buplueri Radix 의 1,2,4-benzentriol에 의해 유발된 DNA Damage에 대한 보호효과에 대한 연구)

  • Lee, Young-Joon;Kang, Su-Jin
    • Journal of Society of Preventive Korean Medicine
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    • v.12 no.2
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    • pp.51-59
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    • 2008
  • Objectives : Buplueri Radix (BR), used medical plant in Korea traditional medicine, contains various compounds, including a series of triterpene saponins known as saikosaponins. We performed this study for the protective effect of BR against oxidative damage induced by 1,2,4-benzentriol(BT) in human lymphocytes. Methods : In order to investigate the protective effect of BR against carcinogens, genotoxicity induced by benzene metabolite, BT were performed using cytokinesis-block micronucleus(CBMN) assay and comet assay. Results : The frequency of micronucleus at 25, 50 and $100{\mu}M$ concentration of BT were $8{\pm}2.36$, $23{\pm}2.31$, $35{\pm}4.17$ respectively. In addition of BR with concentration of 25 and $50{\mu}g/mL$, MN frequencies were significantly decreased. According to comet assay, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 while BT with BR treatment decreased DNA breakage. No genotoxicity was observed by BR($25{\sim}50{\mu}g/mL$) treatment alone on DNA breakage. Since BT can induce DNA damage through the generation of reactive oxygen species(ROS), we examined the level of ROS in human lymphocytes treated with BT and/or BR using DCF-DA, ROS-sensitive probe. The generation of ROS in BT-treated cells was also observed, and BR addition inhibited the level of BT-induced DNA damage. Conclusions : From above results it is suggested that BR could protect the cell and DNA from pro-oxidant effect by ROS by BT

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Comparison of the Sensitivity of Two Micronucleus Assays for Detection of Micronucleus Induction by Cigarette-Smoke Condensate (담배연기응축물의 소핵생성 측정시 두가지 방법간의 민감성 비교)

  • Sohn Hyung-Ok;Lee Young-Gu;Han Jung-Ho;Hur Jae-Yeon;Lee Dong-Wook;Hyun Hak-Chul;Shin Han-Jae
    • Journal of the Korean Society of Tobacco Science
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    • v.26 no.2
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    • pp.152-158
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    • 2004
  • Among short-term in vitro genotoxicity assays, micronucleus assays are rapid, inexpensive, and less labor-intensive system. We have undertaken a comparative study of sensitivity of cigarette smoke condensate(CSC) by general micronucleus(MN) assay and cytokinesis-block micronucleus(CBMN) assay. In this study, V79 Chinese hamster cells were employed to evaluate and compare the genotoxicity of CSC of Kentucky Reference Cigarette 2R4F by 2 kinds of in vitro MN assay methods. To determine the optimum concentration of cytochalasin B(CYB) to obtain the maximal number of binucleated cells for CBMN assay, triplicate cultures of growing cells were treated with CYB for 15 h. CYB treatments caused a concentration-dependent increase in cytotoxicity($1\~4{\mu}g/mL$) and proportion($0.25\~1\;{\mu}g/mL$) of binucleated cells. These data suggested that 1 ug/mL of CYB is as an optimum dose for CBMN assay in binucleated V79 cells. Short treatment(4 h) of CSC induced a micronucleated cells with a concentration-dependent response in the presence or absence of CYB, but CSC-induced MNs were weakened when S9 was present. Long treatments(19 h) of CSC also induced a significant increase MN formation with a concentration-dependent response. At a concentration of 75 ${mu}g/mL$, the MN cell frequencies of general MN assay and CBMN assay were $6.5\%\;and\;11.7\%$, respectively. Linear regression analysis revealed a good correlation in CBMN assay between a concentration of CSC and MN cell frequency. All these data indicated that CBMN assay is more sensitive to the induction CSC-induced MN than general MN assay.

Increased Frequency of Micronuclei in Binucleated Lymphocytes among Occupationally Pesticide-exposed Populations: A Meta-analysis

  • Yang, Hai-Yan;Feng, Ruo;Liu, Jing;Wang, Hai-Yu;Wang, Ya-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6955-6960
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    • 2014
  • Background: The cytokinesis-block micronucleus (CBMN) assay is a standard cytogenetic tool employed to evaluate chromosomal damage subsequent to pesticide exposure. Objectives: To evaluate the pooled levels of total micronuclei (MN) and binucleated cells with micronuclei (MNC) in 1000 binucleated lymphocytes among population occupationally exposed to pesticides and further determine the more sensitive biomarker of CBMN. Materials and Methods: A meta-analysis on the pooled levels of MN and MNC in binucleated lymphocytes among occupationally pesticide-exposed populations was conducted using STATA 10.0 software and Review Manager 5.0.24 in this study. Results: We found significant differences in frequencies of MN and MNC in 1000 binucleated lymphocytes between pesticide-exposed groups and controls, and the summary estimates of weighted mean difference were 6.82 [95% confidence interval (95% CI): 4.86-8.78] and 5.08 (95% CI: 2.93-7.23), respectively. However, when we conducted sensitivity analyses further, only the MN remained statistically different, but not the MNC, the summary estimates of weight mean difference were 2.86 (95% CI: 2.51-3.21) and 0.50 (95% CI: -0.16-1.17), respectively. We also observed pesticide-exposed subjects had significantly higher MN frequencies than controls among smokers and nonsmokers, male and female populations, and American, Asian and European countries in stratified analyses. Conclusions: The frequency of MN in peripheral blood lymphocytes might be a more sensitive indicator of early genetic effects than MNC using the CBMN assay for occupationally pesticide-exposed populations.

Genotoxic and Anti-Genotoxic Effects of Vanillic Acid Against Mitomycin C-Induced Genomic Damage in Human Lymphocytes In Vitro

  • Erdem, Merve Guler;Cinkilic, Nilufer;Vatan, Ozgur;Yilmaz, Dilek;Bagdas, Deniz;Bilaloglu, Rahmi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4993-4998
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    • 2012
  • Vanillic acid, a vegetable phenolic compound, is a strong antioxidant. The aim of the present study was to determine its effects on mitomycin C-induced DNA damage in human blood lymphocyte cultures in vitro, both alone and in combination with mitomycin C (MMC). The cytokinesis block micronucleus test and alkaline comet assay were used to determine genotoxic damage and anti-genotoxic effects of vanillic acid at the DNA and chromosome levels. MMC induced genotoxicity at a dose of $0.25{\mu}g/ml$. Vanillic acid ($1{\mu}g/ml$) significantly reduced both the rates of DNA damaged cells and the frequency of micronucleated cells. A high dose of vanillic acid ($2{\mu}g/ml$) itself had genotoxic effects on DNA. In addition, both test systems showed similar results when tested with the negative control, consisting of dimethyl sulfoxide (DMSO) in combination with vanillic acid ($1{\mu}g/ml$)+MMC. In conclusion, vanillic acid could prevent oxidative damage to DNA and chromosomes when used at an appropriately low dose.

Analysis of radiation-induced micronuclei and aneuploidy involving chromosome 1 and 4 by FISH technique (FISH 기법을 이용한 방사선에 의한 소핵과 이수성 분석)

  • Chung, Hai-Won;Kim, Tae-Yon;Cho, Yoon-Hee;Kim, Su-Young;Kang, Chang-Mo;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.29 no.4
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    • pp.243-249
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    • 2004
  • The cytokinesis-block micronucleus (CBMN) assay in combination with FISH technique using chromosome-specific centromeric probes for chromosome 1 and 4 was performed in mitogen stimulated human lymphocytes which were exposed to x-radiation to identify different sensitivity of chromosomes to the induction of micronuclei(MN) and aneuploidy by radiation. The frequencies of micronucleated cytokinesis-blocked(MNCB) cells and MN in binucleated lymphocytes(BN) increased with the increase in radiation dose. A significant induction of aneuploidy of chromosome 1 and 4 were found. The frequency of aneuploidy of chromosome 1 and 4 in the control were 9 per 2,000 BN cells and this increased to 47 and 71 following irradiation at a dose of 1 and 2 Gy, respectively. The induction of aneuploidy of chromosome 1 was higher than that of chromosome 4. The frequency of aneuploid BN cells with MN exhibiting positive centromere signal for either chromosome 1 and/or 4 increased in a dose dependent manner, and that for chromosome 1 is higher than that for chromosome 4. Among the total induced MN in irradiated lymphocytes, smaller proportion of MN exhibit centromeric signal of chromosome indicating that radiation-induced MN are mainly originated from chromosomal breakage rather than chromosomal non-disjunction. These results suggest that x-radiation can induce aneuploidy and supports the finding that chromosome vary in their sensitivity to aneuploidy induction by x-irradiation.

Cytotoxicity in HL-60 cells and human lymphocytes and effect of leukemia cell differentiation induced by Saussureae Radix extract (정상사람림프구와 HL-60 cell에서 목향의 세포독성과 백혈병세포 분화효과에 관한 연구)

  • Lee, Young-Joon;Kang, Su-Jin;Ku, Sae-Kwang
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.31-37
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    • 2011
  • Objectives : This study was focused to investigate the toxicity of Saussurea lappa (SL) extracts in HL-60 cells and human lymphocytes. We also examined the differentiation effect of SL against leukemia cells. Methods : For examining the toxicity of SL, cytokinesis-block micronucleus (CBMN) assay and single cell gel eletrophoresis (SCGE) assay were used in present study. The cell differentiation effect of SL was evaluated by nitroblue tetrazolium (NBT) reduction assay. Results : The inhibition of cell growth in HL-60 cells was observed in a dose-dependant manner after SL treatment for 24 h. According to SCGE assay, HL-60 cells treated with SL increased DNA damage at $10{\mu}g/m{\ell}$, while DNA damage was induced by 0.1, 1, $10{\mu}g/m{\ell}$ concentration of SL in human lymphocytes. Our results indicated that SL have no genotoxic effect in HL-60 cells and human lymphocytes. Additionally, the differentiation effect was induced in $1{\mu}g/m{\ell}$ SL-treated HL-60 cells. Conclusions : From above results it is suggested that SL could be beneficial for the preparation of the useful agent for treating leukemia.

Analysis of Chromosome Aberrations and Micronuclei in Peripheral Lymphocytes of Nurses in a Hospital (병원 근무 간호사들의 염색체 이상 및 소핵 빈도에 미치는 위험요인)

  • Jeon Hye Jeong;Kim Tae Hwan;Choi Soo Yong;Chung Hai won
    • Environmental Mutagens and Carcinogens
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    • v.25 no.4
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    • pp.150-156
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    • 2005
  • Analysis of chromosome aberration (CA) and cytokinesis-block micronucleus (CBMN) assay in peripheral lymphocytes of nurses exposed to low levels of anticancer drug and Ethylene Oxide(EO) gas in a hospital were performed. The frequency of CA was increased in the exposed compared to the controls whereas no increase of the frequency of MN was found. The frequencies of chromatid type CA were 1.2, 3.91 and 9.67 per 500 cells in the controls, workers exposed to anticancer drug and workers exposed to EO, respectively. Lower frequency of CA in nurses handling anticancer drugs with safety covers compared to those without safety covers was observed, but it was not statistically significant. The frequency of CA in nurses handling anticancer drugs increased by the frequency of mixing anticancer drugs. Poisson regression analysis showed a significant association of the frequency of chromatid type CA with age, duration of wort exposure to anticancer drug and EO gas exposure, but no association of the frequency of chromosome type CA with any variables. The results suggested that there were associations between CA and the occupational exposure to low levels of anticancer drug and EO gas.

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