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Kim, Youn-Jung,Ryu, Jae-Chun The Korean Society of Toxicogenomics and Toxicopro 2007 Molecular & cellular toxicology Vol.3 No.2
Chlorobenzenes due to their acute toxicity and the capability of bioaccumulating are of great health and environmental concern. Especially, 1, 2-dichlorobenzene (CAS No. 95-50-1) is used for organic synthesis, dye manufacture, as a solvent and for other applications in chemical industry. Adverse effects of 1, 2-dichlorobenzene includes increases in liver and kidney weights and hepatotoxicity. In this study, we evaluated the genetic toxicity of 1, 2-dichlorobenzene with more advanced methods, in vitro mouse lymphoma assay $tk^{+/-}$ gene assay (MLA) and in vitro mouse supravital micronucleus (MN) assay. 1, 2-Dichlorobenzene appeared the significantly positive results and the induction of large mutant colonies only in the presence of metabolic activation system with MLA. But in vitro testing of 1, 2-dichlorobenzene yielded negative results with supravital MN assay. These results suggest that 1, 2-dichlorobenzene may play a mutagen rather than clastogen in vitro mammalian system.
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Kim, Youn-Jung,Ryu, Jae-Chun The Korean Society of Toxicogenomics and Toxicopro 2007 Molecular & cellular toxicology Vol.3 No.3
Butylated hydroxytoluene (BHT) is widely used antioxidant food additives. It has been extensively studied for potential toxicities. BHT appears adverse effects in liver and thyroid. In this study, we evaluated the genetic toxicity of BHT with more advanced methods, in vitro mouse lymphoma assay $tk^{+/-}$ gene assay (MLA) and in vivo mouse supravital micronucleus (MN) assay. BHT did not appear the significantly results in the absence and presence of metabolic activation system with MLA. Also, in vivo testing of BHT yielded negative results with supravital MN assay. These results suggest that BHT itself was not generally considered genotoxic.
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김종원,한의식,박미선,엄미옥,전혜승,정해관,오혜영 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-
의약품등록의 국제적합의를 위한 협의체(IC딨 . Interna금onal Conference on Harmonization of TecHnological Requirements for Registration of fharrnaceuticats for Human Use)에서 제시하는 유전폭성시험법에서 검색되지 않는 비유전독성 발암물질을 위한 새로운 시험법을 개발하기 위하여 1차년도 연구에서 이러찬 물질에 대한 검색 가능성을 나타낸 '생체외 소핵.시험'과 최근 환경변이원학계에서 새로이 그 유용성이 퍼두되고 있는 'Syrian hamster embryo (SHE) 세포형질전환 시험법'을 이용하여 알려진 두가지 좋류의 비유전독성 발암물질의 반응을 연구하였다. 생체외 소핵시험에서는 2,3,7,8-tetrachlorodibenzo-p-dioxin fTCDD)를 모델화합물로 선정하고, 사람유방 유래의 MCF-7, MCF-lOA,MDA-MB-231 세포를 사용하여 에스트로젠 수용체(ERf 존재여부와 세포의 유형에 파른 소핵형성을 연구하였으며, 등시에 에스트로젠 길항제인 타목시펜에 대한 영향을 보았다. 또한 TCDD가 MCF극 세포의 세포주기예 미치는 영향을 연구하기 위하여 소핵형성을 니-타렌 농도에서 flow cytometer를 이용한 세포주기 분석을 수행하였다. TCDD는 MCF-lOA세포 (ER negative)와 MDA-MB-231세포 (ER negative)에서는 소핵형성이 디미하거나 전혀 증가하지 않았으나, 뽀Cf-7세포 (ER positive)에서 재현성있는 소핵형성을 유발하였으며, 타목시펜 (10 nUt)은 TCDD (IpM)에 의한 소핵형성을 최고 47.3% 까지 억제하였다. 근또글 세포주기분석결과 TCDD, tamoxi(on 단독처리군은 읍성대조군과 유사하몄으나,병용처리군에서는 GO#'Gl arrest를 유의성있게 유발하였다. SHE 세포 형질전환시험법에서는 1차년도 연구에서 생체외 소핵시험에서 양성의 결과를 나타낸 bisphenoB A을 모델화합물로 하여 연구하였다. 시험조건하에서 blsphenol A(31.2SfM, 48시간 연속처리)는 세포혈질전찬을 유발하였다. 상기 연구결과로부터 비유전독성발암물질은 기존의 유전독성시험법중 생체왹 소핵시험(에스트로젠 수용체 양성인 세포주)과 세포형질전환#·1험(Syrian hamster 배자 초대배양세포)에서 시험계의 선택 여부에 따라 부분적으로. 유전독성 지표를 검색할 수 있음을 증명하였으며, 향후 본 연구결과에서 양성을 나타낸 세포의 세포주기 관련인자의 발현변확 등에 대한 작용기전 연구가 진행되어 새로운 생체지표를 이용한 시험법의 개발이 될 것이다 To identify nongenotoxic carcinogens which are not indentified by ICH (Internation Harmonization of I'echnological Requirements ·for Registration of Pharmaceuticals for Human Use) guideline-recommended standard genotoricfty test battery, two endpoints were chosen. They are in vitro micronilcleus 3ss3y which was useful to indentify for bisphenol Aand DEHP last year's study and in vitro 51'rian hamster embryo (SHE) ceBl transformationassay which was recently recognized as a useful tool to detect nongenotoxic carcinogen as wellas genotoxic carcnogen by Environmetal Mutagen Society. In the in vitro micronucleus assayusing MCF-7 cells, TCBD is all negative in standard genotoxicity test battery; Ames test,chromosome abem·ation assay, mouse 3ymphoma tk+/- assay, in vivo micronuclelts assay.therefore TCDD if picked up as a model compound- TCDD atso induced micronucleusformation. To identify the relationship between MN formation and estrogen receptor (ER),TCDO, BPA, and DEHP were studied using hfCF-7 cells (ER positive) and in MCF-tOfL cells(ER negative), MDA-MB-231'ce31s (ER negative). TCDD induced MN formation in bfCF-7celts (ER positive) but not or little induced MN fornlation in MCF-lOA cells (ER negative) andMDA-MB-231 (ER negative). BPA and DEHP didn't induced MN formation in MCF-lOA cells(ER negative). NP didn't induce MN formation in CHt cells but induced MN formation inMCF-7 cells, We also have included the effect of estrogen inhibitors, tamoxifen, against TCDDinduced MN formation. Tamoxifen inhihited TCDD-induced UtN formation up to 4f.3% in,4fCF-7 cells. in cell cycle analysis using flow cytorneter, 1'CDD (1 pM), tamcrifen (10 nM)didn't change cell cycle phase compared with that of negative control but, tamoxifen (10 nhf)with TCDD (1 pM) induced GO/Gl arrest significant]y. Inferring for the result above 4compounds (BPA, EIEHP, TCDD, NP), we concluded the in vitro micronucleus assay vsingMCF-7 cells is a good test method for detecting nongenotoxic carcinogen and identifying thegenotoxicity of endocrine distruptors. In the in vitro SHE celt transformation assay, bisphenol Ais picked up as a model compound. Treated for 48 hr, Bisphenol A induced morphotogicaltransformation significantly.'Strategy and methological approach for identification ofnonBenotoxic cartifogen in vitro'That to identify nongenotcxic carcinogens wllich are notidentified by standard genetic toxicology test battery should be included with additive endpointsIs convincing.
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유전독성 대체시험법(in vitro소핵시험)의 국내검증시험연구
이현걸 ( Hyun Kul Lee ),김종극 ( Jong Geuk Kim ),이우주 ( Woo Joo Lee ),맹은호 ( Eun Ho Maeng ),이종윤 ( Jong Yoon Lee ),정영신 ( Young Shin Chung ),김창환 ( Chang Hwan Kim ),김윤순 ( Yun Soon Kim ),장미해 ( Mi Hae Jang ),이석종 ( 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1
This validation study was performed to introduce and set up the in vitro micronucleus test, as an alternative to the existing chromosome aberration test, in the test facilities in Korea, and to establish the competence of each facility to perform the in vitro micronucleus test independently. The 10 coded substances, including 6 positive and 4 negative in genotoxicity assay, were tested. Six facilities were divided into two subgroups, A and B. Three coded positive compounds and two negative compounds were sent to each test facility. The compounds were tested according to the facility`s GLP system. In group A, positive results were obtained from testing of the three positive compounds, and negative results were obtained from testing of the two negative compounds in all three facilities. In group B, positive results were also obtained from testing of the three positive compounds, and negative results were obtained from testing of the two negative compounds in all three facilities. For the Urethane, an in vitro equivocal compound tested by group B, negative results were obtained. These results suggested that the compound may be effectively activated only in in vivo environment. The results show that the six test facilities are capable of performing the in vitro micronucleus test independently and producing consistent results.
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연구논문 : 유전독성 대체시험법(in vitro소핵시험)의 국내검증시험연구
이현걸 ( Hyun Kul Lee ),김종극 ( Jong Geuk Kim ),이우주 ( Woo Joo Lee ),맹은호 ( Eun Ho Maeng ),이종윤 ( Jong Yoon Lee ),정영신 ( Young Shin Chung ),김창환 ( Chang Hwan Kim ),김윤순 ( Yun Soon Kim ),장미해 ( Mi Hae Jang ),이석종 ( 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1
This validation study was performed to introduce and set up the in vitro micronucleus test, as an alternative to the existing chromosome aberration test, in the test facilities in Korea, and to establish the competence of each facility to perform the in vitro micronucleus test independently. The 10 coded substances, including 6 positive and 4 negative in genotoxicity assay, were tested. Six facilities were divided into two subgroups, A and B. Three coded positive compounds and two negative compounds were sent to each test facility. The compounds were tested according to the facility`s GLP system. In group A, positive results were obtained from testing of the three positive compounds, and negative results were obtained from testing of the two negative compounds in all three facilities. In group B, positive results were also obtained from testing of the three positive compounds, and negative results were obtained from testing of the two negative compounds in all three facilities. For the Urethane, an in vitro equivocal compound tested by group B, negative results were obtained. These results suggested that the compound may be effectively activated only in in vivo environment. The results show that the six test facilities are capable of performing the in vitro micronucleus test independently and producing consistent results.
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DK1002에 대한 급성독성시험 및 유전독성에 관한 연구
류재천,김경란,김현주,정상운,김명국,박희석,김용해,Ryu, Jae-Chun,Kim, Kyung-Ran,Kim, Hyun-Joo,Jung, Sang-Oun,Kim, Myung-Kuk,Park, Hee-Sock,Kim, Yong-Hae 한국독성학회 1998 Toxicological Research Vol.14 No.3
The acute and genetic toxicity of DK1002 was subjected in this study. DK1002 which is a morphine-like new drug candidate synthesized by Dong-Kook Pharmaceutical Co. Ltd. is now under developing as a analgesics that have better drug efficacy and least addictive property. In acute toxicity study, the 50% lethal doses ($LD_{50}$) of DK1002 were determined as>2000mg/kg (p.o.), 237.0mg/kg(i.p.), 57.5mg/kg(i.v.), and 1266.9mg/kg (s.c.). And also, to study the genotoxicity of DK1002, we performed bacterial reversion assay with Salmonella typhimurium TA98, TA100, TA1535, and TA1537, and in vitro chromosomal aberration assay with Chinese hamster lung cells in the presence and absence of S-9 metabolic activation system. In vivo micronucleus assay using mouse bone marrow cells was also performed. From these results, DK1002 was revealed nonmutagenic potential in S. typhimurium TA98, TA100, TA1535, and TA537 both in the absence and presecne of metablic activation system. No clastogenicity of DK1002 was observed in chromosomal aberration assay in vitro as well as in micronucleus assay in vivo.
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