• 제목/요약/키워드: Micronuclei assay

검색결과 75건 처리시간 0.019초

마우스와 사람 림프구에서 방사선에 의한 미소핵의 형성 및 고려인삼의 효과 (Induction of Micronuclei in Human and Mouse Lymphocytes Irradiated with Gamma Radiation and Effect of Panax ginseng C.A. Meyer)

  • 김성호;오헌;이송은;이윤실;김태환;정규식;류시윤
    • Journal of Radiation Protection and Research
    • /
    • 제22권3호
    • /
    • pp.153-160
    • /
    • 1997
  • 사람의 말초혈액림프구와 C57BL/6마우스의 비장림프구를 사용하여 시험관내에서 감마선을 조사하고 배양하여 세포질분열 차단 림프구내에 형성되는 미소핵의 빈도를 측정하였다. 미소핵 발생빈도는 방사선조사 선량에 비례하여 증가하였으며 linen-quadratic 곡선식에 적용하여, 세포 당 0.2개의 미소핵이 유도되는 방사선량을 산출하면 사람의 말초혈액 림프구에 비하여 마우스 비장림프구에서 1.67배 민감하였다. 미소핵시험방법을 이용하여, 사람의 말초혈액 림프구에 대한 인삼의 방사선 방호효과를 시험관내 시험으로, 마우스의 미장림프구에 대한 효과를 생체내 시험으로 검정하였다. 사람림프구에 있어서 방사선(3Gy)에 의해 유도되는 미소핵의 수는 방사선조사 전 및 후 투여군에서 공히 감소하였으며(p<0.01), 마우스를 사용한 생체시험에서도 림프구의 미소핵 발생빈도는 낮게 관찰되었다(p<0.025). 이상의 결과에서 인삼은 인체에서도 방사선에 의한 세포장해를 감소시킬 가능성을 나타냈다.

  • PDF

DNA 회복 저해제 Cytosine Arabinoside, 3-Aminobenzamide 및 Hydroxyurea가 방사선에 의해 유도된 소핵과 이수성에 미치는 영향 (Effect of Cytosine Arabinoside, 3-Aminobenzamide and Hydroxyurea on the frequencies of radiation-induced micronuclei and aneuploidy in human lymphocytes)

  • 조윤희;김양지;강창모;하성환;정해원
    • Journal of Radiation Protection and Research
    • /
    • 제30권4호
    • /
    • pp.209-219
    • /
    • 2005
  • 소핵분석은 방사선의 생물학적 선량계로서 활용되고 있으나 이의 생성 기전은 아직까지 확실치 않다. 본 연구에서는 사람 림프구에 방사선을 조사한 후 DNA 손상회복 저해물질, Cytosine Arabinoside(Ara C)와 3-Aminobenzamide(3-AB) 그리고 Hydroxyurea(HU)를 특정 세포주기에 처리하고 소핵분석과 FISH기법을 이용하여 방사선에 의한 소핵 및 이수성의 정도를 구명하고자 하였다. 방사선 선량에 따라 소핵과 이수성의 빈도는 양반응 관계를 보이며 증가하였고 DNA 손상회복 저해물질 처리 후 소핵의 빈도는 모든 DNA 손상 회복 저해물질에 의해 증가하였으며 Ara C, 3AB, HU 순으로 나타났다. 이수성의 빈도는 HU와 Ara C에 의해서 크게 증가하였으나, 3AB는 아무런 영향을 주지 않았다. 또한 1번 염색체가 4번 염색체보다 방사선에 의한 소핵형성 및 이수성에 더 많이 관여되었다. 본 연구 결과, 방사선에 의한 소핵 및 이수성의 형성 과정은 여러 다른 기전이 관여하고 있음을 알 수 있었다.

사람, 토끼 및 개 유래 말초혈액 림프구의 미소핵을 이용한 방사선 피폭의 생물학적 선량측정 (Induction of micronuclei in human, rabbit and dog lymphocytes irradiated in vitro with gamma radiation)

  • 류시윤;강빛나;김호준;김태환;정규식;김세라;이해준;김성호;안미영
    • 대한수의학회지
    • /
    • 제42권1호
    • /
    • pp.1-6
    • /
    • 2002
  • The frequencies of gamma-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of three species (human, rabbit, dog). Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. When analysed by linear-quadratic model the line of best fit was : human : $y=0.1184D+0.01867D^2+0.01$, rabbit : $y=0.0387D+0.00528D^2+0.01$ (y = number of MN/CB cells and D = irradiation dose in Gy). The relative sensitivity of rabbit lymphocytes compared with human lymphocytes was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 Gy to 4 Gy. In the case of MN frequency with 0.2, the relative sensitivities of rabbit lymphocytes was 0.39. These data indicate that the induction of MN in rabbit CB cells following irradiation was much less sensitive to the MN induction effects of gamma-irradiation than those from human. The MN assay with dog lymphocytes was very difficult and time-consumed because the dog PHA-stimulated lymphocytes yielded cultures with very low level of CB cells formation in the condition of this experiment. Our in vitro radiobiological study confirmed that the cytogenetic response obtained in blood from rabbit can be utilized for application in environmental studies.

Peripheral Blood Lymphocytes as In Vitro Model to Evaluate Genomic Instability Caused by Low Dose Radiation

  • Tewari, Shikha;Khan, Kainat;Husain, Nuzhat;Rastogi, Madhup;Mishra, Surendra P;Srivastav, Anoop K
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제17권4호
    • /
    • pp.1773-1777
    • /
    • 2016
  • Diagnostic and therapeutic radiation fields are planned so as to reduce side-effects while maximising the dose to site but effects on healthy tissues are inevitable. Radiation causes strand breaks in DNA of exposed cells which can lead to chromosomal aberrations and cause malfunction and cell death. Several researchers have highlighted the damaging effects of high dose radiation but still there is a lacuna in identifying damage due to low dose radiation used for diagnostic purposes. Blood is an easy resource to study genotoxicity and to estimate the effects of radiation. The micronucleus assay and chromosomal aberration can indicate genetic damage and our present aim was to establish these with lymphocytes in an in vitro model to predict the immediate effects low dose radiation. Blood was collected from healthy individuals and divided into 6 groups with increasing radiation dose i.e., 0Gy, 0.10Gy, 0.25Gy, 0.50Gy, 1Gy and 2Gy. The samples were irradiated in duplicates using a LINAC in the radiation oncology department. Standard protocols were applied for chromosomal aberration and micronucleus assays. Metaphases were stained in Giemsa and 200 were scored per sample for the detection of dicentric or acentric forms. For micronuclei detection, 200 metaphases. Giemsa stained binucleate cells per sample were analysed for any abnormality. The micronuclei (MN) frequency was increased in cells exposed to the entire range of doses (0.1-2Gy) delivered. Controls showed minimal MN formation ($2.0%{\pm}0.05$) with triple MN ($5.6%{\pm}2.0$) frequency at the lowest dose. MN formation increased exponentially with the radiation dose thereafter with a maximum at 2Gy. Significantly elevated numbers of dicentric chromosomes were also observed, even at doses of 0.1-0.5Gy, compared to controls, and acentric chromosomes were apparent at 2Gy. In conclusion we can state that lymphocytes can be effectively used to study direct effect of low dose radiation.

Genetic Toxicity Test of 8-Hydroxyquinoline by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Woo-Sun;Kim, Hyun-Joo;Lee, Eun-Mi;Kim, Joo-Hwan;Suh, Soo-Kyung;Kwon, Kyung-Jin;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-N.
    • Molecular & Cellular Toxicology
    • /
    • 제3권2호
    • /
    • pp.90-97
    • /
    • 2007
  • 8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.

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

  • 정해원;김태연;조윤희;김수영;강창모;하성환
    • Journal of Radiation Protection and Research
    • /
    • 제29권4호
    • /
    • pp.243-249
    • /
    • 2004
  • 본 연구는 소핵분석과 염색체 1번 및 4번의 DNA probe를 이용한 FISH 기법을 병행하여 방사선에 의한 소핵과 이수성에 관여하는 각 염색체의 감수성을 평가하고자 하였다. 방사선 선량에 따라 소핵의 빈도는 증가하였으며 염색체 1번과 4번의 이수성도 대조군, 1 Gy 및 2 Gy 에서 각각 2000개의 BN세포 당 9개, 47개 및 71개로 유의하게 증가하였다. 염색체 1번의 이수성 빈도는 4번에 비해 높게 관찰되었다. 염색체 1번 및 4번을 포함하는 소핵도 방사선의 선량에 따라 증가하였으며, 소핵내 염색체 1번의 포함빈도가 4번보다 높게 관찰되었다. 또한 방사선에 의한 소핵 중 낮은 빈도의 염색체 signal를 포함하는 소핵이 관찰됨으로써 방사선에 의한 소핵은 대부분 절단에 의한 것임을 확인할 수 있었다. 따라서 본 연구 결과 방사선은 이수성을 유도하며 이에 염색체가 다르게 관여할 수 있음을 보여준다.

Effects of Flavonol Derivatives on the Micronudei Formation by N-methyl-N'-nitro-N-nitrosoguanidine and the Enhancement of Bleomycin-induced Chromosome Aberration by N-methyl-N'-nitro-N-nitrosoguanidine

  • Heo, Moon-Young;Kwon, Chang-Ho;Sohn, Dong-Hun;Lee, Su-Jun;Kim, Sung-Wan;Kim, Jung-Han;William W. Au
    • Archives of Pharmacal Research
    • /
    • 제16권3호
    • /
    • pp.196-204
    • /
    • 1993
  • Flavonol derivatives were tested for their anticlastogenic effect against induction of micronuclei by n-methyl-n'-nitor-n-nitorsoguanidine(MNNG), and against induction of chromosome aberration by bleomycin or MNNG.belomycin. For micronudeus assay, each flavonol derivative (0, 0.001, 0.01, 0.1, 1, 10 and 100 mg/kg) was administered orally twice with 24 h interval, together with intraperitioneally administered MNNG(150 mg/kg). The result showed that msot flavonol derivatives tested were effective in suppresing the frequencies of micronude induced by MNNG. For chromosome aberration assy, each flavonol derivative (0, 0.1, 1, 10m and 100 mg/kg)was administered to mice orally in vivo, and then mice were sacrificed and spleen lymphocyte cultures were made. Bleomycin $(3\;\mu$g/ml) was treated to the mouse spleen hymphocyte cultures at 24 h after con A initiation. There wre nomarked decrease tendencies in chromosome aberration unless all doses of galangin and some doses of several flavonol derivatives tested. In the another experiment, we have evaluated the effect of flavonol derivatives on the enhancement of bleomycin-induced chromsome aberration by MNNG. Most of flavonol derivtives reduced the incidence of chromosome aberration induced by in vitro treatment of bleomycin followed by in vivo treatment of MNNG. Galangin particulary showed a dose-dependent decrease tendency. Other flavonol derivative showed slightly suggest that most of flavonol derivatives may be capable of protecting the inhibition of suggest that most of flavonol derivatives may be capable of protecting the inhibition of DNA-repair by MNNG. Our data indicate clearly that flavonol derivatives can suppress MNNG-induced genotoxicity such as an induction of MNPCEs. Therfore, our results could suggest that flavonol derivtives may be useful as a chemopreventive agent of MNNG.

  • PDF

Genetic Toxicity Test of Glycidol by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ji-H.;Kim, Ki-Y.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Sheen, Yhun-Y.
    • Biomolecules & Therapeutics
    • /
    • 제14권4호
    • /
    • pp.240-245
    • /
    • 2006
  • The primary use for glycidol is as a stabilizer in the manufacture of vinylpolymers, however, it is also used as an intermediate in the production of pharmaceuticals, as an additives for oil and synthetic hydraulic fluids, and as a diluting agent is same epoxy resins. In this study, we have carried out in vitro genetic toxicity test of glycidol and microarray analysis of differentially expressed genes in response to glycidol. The result of Ames test showed mutations with glycidol treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, glycidol showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with glycidol treatment showed DNA damage both with and without exogenous metabolic activation. Glycidol increased micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to glycidol by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of glycidol.

고선량 감마선조사 곡류 분말의 유전독성학적 안전성평가 (Genotoxicological Safety of High-Dose Gamma-Irradiated Cereal Powders)

  • 한삭명;김혜미;정승경;이주운;변명우;강일준
    • 한국식품저장유통학회지
    • /
    • 제13권4호
    • /
    • pp.524-529
    • /
    • 2006
  • 곡류분말의 위생화 및 물리적 특성 개선을 위해 감마선 조사기술의 이용 가능성이 높아짐에 따라 이들의 안전성을 확보할 목적으로 30 kGy의 고선량 감마선조사 곡류분말의 유전독성학적 안전성 평가를 실시하였다. 감마선 조사 및 비조사 곡류분말의 S. typhimurium TA 98, TA100, TA1535 및 TA1537에 대한복귀변이 집락수를 조사한 결과, 대사활성계 도입 및 부재시 모두 시험적용 농도인 0.625-10 mg/plate의 범위에서 복귀변이 집락수의 농도 의존적인 증가 혹은 감소를 보이지 않아 감마선 조사 곡류분말(30 kGy)은 돌연변이원성이 없는 것으로 나타났다. 또한, 설치류 망상적혈구를 이용하여 감마선조사 곡류분말의 소핵 형성 시험을 수행한 결과, 30 kGy 감마선 조사 곡류분말은 시험적용 용량인 625-5,000 mg/kg의 범위에서 소핵을 가진 망상 적혈구의 출현율이 음성대조군과 유의한 차이를 나타내지 않아 소핵을 유발하지 않음을 확인하였다. 포유류 배양세포를 이용한 염색체 이상시험에서도 30 kGy 감마선 조사곡류분말은 시험적용 용량에서 염색체이상 유발능이 5% 미만이었다.

Lactobacillus plantarum AF1와 Lactobacillus plantarum HD1이 생성한 조항균 물질의 유전학적 독성평가 (A Genotoxicological Safety Evaluation of Crude Antifungal Compounds Produced by Lactobacillus Plantarum AF1 and Lactobacillus Plantarum HD1)

  • 장해춘;고상범;이재준
    • 한국지역사회생활과학회지
    • /
    • 제26권4호
    • /
    • pp.633-645
    • /
    • 2015
  • This study investigates the genotoxicity of crude antifungal compounds produced by Lactobacillus plantarum AF1 (L.plantarum AF1) and Lactobacillus plantarum HD1 (L. plantarum HD1) isolated from kimchi. The genetic toxicity of crude antifungal compounds was evaluated in bacterial reverse mutation in Salmonella and Escherichia spp., chromosome aberrations in Chinese hamster lung cells, and micronucleous formations in mice. In bacterial reversion assays with Salmonella Typhimurium TA98, TA100, TA1535, TA1537, and WP2uvrA, crude antifungal compounds did not increase the number of revertant colonies in both the absence and presence of the 59 metabolic activation system. In the chromosome aberration test with Chinese hamster lung cells, crude antifungal compounds showed no increase in the frequency of chromosome aberrations in the short-period test with/without the S9 mix or in the continuos test. In the in vivo mouse micronucleus assay, crude antifungal compounds showed no increase in the frequency of polychromatic erythrocytes with micronuclei. The results show that crude antifungal compounds produced by L. plantarum AF1 and L. plantarum HD1 did not induce any genotoxicity.