In vivo DNA mismatch repair measurement in zebrafish embryos and its use in screening of environmental carcinogens

doi:10.1016/j.jhazmat.2015.09.056

科研成果详情

题名	In vivo DNA mismatch repair measurement in zebrafish embryos and its use in screening of environmental carcinogens
作者	Chen, Yuanhong1; Huang, Changjiang 1; Bai, Chenglian1; Du, Changchun 1; Liao, Junhua1; Dong, Qiaoxiang1,2
发表日期	2016-01-25
发表期刊	JOURNAL OF HAZARDOUS MATERIALS 影响因子和分区
语种	英语
原始文献类型	Article
关键词	DNA mismatch repair CdCl2 BaP PFOS Zebrafish embryo Environmental carcinogens
其他关键词	MODEL SYSTEM ; EXTRACTS ; CELLS ; HMLH1 ; HETERODIMER ; CADMIUM ; IDENTIFICATION ; CONSTRUCTION ; RECOGNITION ; INSTABILITY
摘要	Impairment of DNA mismatch repair (MMR) function leads to the development and progression of certain cancers. Many environmental contaminants can target DNA MMR system. Currently, measurement of MMR activity is limited to in vitro or in vivo methods at the cell line level, and reports on measurement of MMR activity at the live organism level are lacking. Here, we report an efficient method to measure DNA MMR activity in zebrafish embryos. A G-T mismatch was introduced into enhanced green fluorescent protein (EGFP) gene. Repair of the G-T mismatch to G-C in the heteroduplex plasmid generates a functional EGFP expression. The heteroduplex plasmid and a similarly constructed homoduplex plasmid were injected in parallel into the same batch of embryos at 1-cell stage and EGFP expression in EGFP positive embryos was quantified at 24h after injection. MMR efficiency was calculated as the total fluorescence intensity of embryos injected with the heteroduplex construct divided by that of embryos injected with the homoduplex construct. Our results showed 73% reduction of MMR activity in embryos derived from MMR-deficient mlh1 mutant fish (positive control) when compared with embryos from MMR-competent wild type AB line fish, indicating feasibility of in vivo MMR activity measurement in zebrafish embryos. We further applied this novel assay for measurement of MMR efficiency in embryos exposed to environmental chemicals such as cadmium chloride (CdCl2), benzo[a]pyrene (BaP), and perfluorooctanesulphonic acid (PFOS) from 6 hpf to 24 hpf. We observed significant reductions of MMR efficiency in embryos exposed to 0.1 mu M CdCl2 (52%) and 0.5 mu M BaP (34%), but no effect in embryos exposed to PFOS. Our study for the first time provides a model system for in vivo measurement of DNA MMR activity at the organism level, which has important implications in risk assessment of various environmental carcinogens. (C) 2015 Elsevier B.V. All rights reserved.
资助项目	National Environmental Protection Public Welfare Science and Technology Research Program of China [201309047]; Natural Science Foundation of Zhejiang ProvinceNatural Science Foundation of Zhejiang Province [Y2110659, LZ13B070001]; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [41271491]
出版者	ELSEVIER
出版地	AMSTERDAM
ISSN	0304-3894
EISSN	1873-3336
卷号	302 页码:296-303
DOI	10.1016/j.jhazmat.2015.09.056
页数	55
WOS类目	Engineering, Environmental ; Environmental Sciences
WOS研究方向	Engineering ; Environmental Sciences & Ecology
WOS记录号	WOS:000366225300035
收录类别	SCIE ; PUBMED ; EI ; SCOPUS
EI入藏号	20154201411801
EI主题词	Risk assessment
URL	查看原文
PubMed ID	26476317
SCOPUSEID	2-s2.0-84944235902
通讯作者地址	[Huang, Changjiang]Institute of Environmental Safety and Human Health,Wenzhou Medical University,Wenzhou,325035,China
Scopus学科分类	Environmental Engineering;Environmental Chemistry;Waste Management and Disposal;Pollution;Health, Toxicology and Mutagenesis
TOP期刊	TOP期刊
引用统计	被引频次：5[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/13360
专题	公共卫生学院_健康与环境生态研究所检验医学院（生命科学学院、生物学实验教学中心）公共卫生学院_环境安全与健康风险评估研究院
通讯作者	Huang, Changjiang
作者单位	1.Institute of Environmental Safety and Human Health,Wenzhou Medical University,Wenzhou,325035,China; 2.School of Laboratory Medicine and Life Science,Wenzhou Medical University,Wenzhou,325035,China
第一作者单位	温州医科大学
通讯作者单位	温州医科大学
第一作者的第一单位	温州医科大学
推荐引用方式 GB/T 7714	Chen, Yuanhong,Huang, Changjiang,Bai, Chenglian,et al. In vivo DNA mismatch repair measurement in zebrafish embryos and its use in screening of environmental carcinogens[J]. JOURNAL OF HAZARDOUS MATERIALS,2016,302:296-303.
APA	Chen, Yuanhong, Huang, Changjiang, Bai, Chenglian, Du, Changchun, Liao, Junhua, & Dong, Qiaoxiang. (2016). In vivo DNA mismatch repair measurement in zebrafish embryos and its use in screening of environmental carcinogens. JOURNAL OF HAZARDOUS MATERIALS, 302, 296-303.
MLA	Chen, Yuanhong,et al."In vivo DNA mismatch repair measurement in zebrafish embryos and its use in screening of environmental carcinogens".JOURNAL OF HAZARDOUS MATERIALS 302(2016):296-303.