胞质分裂阻断微核细胞分析法

Cytokinesis-block micronucleus cytome assay.

作者信息

Fenech Michael

机构信息

Genome Health Nutrigenomics Laboratory, CSIRO Human Nutrition, Food Science Australia, PO Box 10041, Adelaide 5000, South Australia, Australia.

出版信息

Nat Protoc. 2007;2(5):1084-104. doi: 10.1038/nprot.2007.77.

DOI:10.1038/nprot.2007.77

PMID:17546000

Abstract

The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated (BN) cells and include (a) micronuclei (MNi), a biomarker of chromosome breakage and/or whole chromosome loss, (b) nucleoplasmic bridges (NPBs), a biomarker of DNA misrepair and/or telomere end-fusions, and (c) nuclear buds (NBUDs), a biomarker of elimination of amplified DNA and/or DNA repair complexes. Cytostatic effects are measured via the proportion of mono-, bi- and multinucleated cells and cytotoxicity via necrotic and/or apoptotic cell ratios. Further information regarding mechanisms leading to MNi, NPBs and NBUDs formation is obtained using centromere and/or telomere probes. The assay is being applied successfully for biomonitoring of in vivo genotoxin exposure, in vitro genotoxicity testing and in diverse research fields such as nutrigenomics and pharmacogenomics as well as a predictor of normal tissue and tumor radiation sensitivity and cancer risk. The procedure can take up to 5 days to complete.

摘要

胞质分裂阻滞微核细胞分析法是一种用于测量DNA损伤、细胞生长停滞和细胞毒性的综合系统。DNA损伤事件在单次分裂的双核（BN）细胞中进行特异性评分，包括：（a）微核（MNi），染色体断裂和/或整条染色体丢失的生物标志物；（b）核质桥（NPB），DNA错配修复和/或端粒末端融合的生物标志物；（c）核芽（NBUD），扩增DNA和/或DNA修复复合物消除的生物标志物。细胞生长停滞效应通过单核、双核和多核细胞的比例来测量，细胞毒性通过坏死和/或凋亡细胞比率来测量。使用着丝粒和/或端粒探针可获得有关导致MNi、NPB和NBUD形成机制的更多信息。该分析法已成功应用于体内基因毒素暴露的生物监测、体外遗传毒性测试以及营养基因组学和药物基因组学等不同研究领域，以及作为正常组织和肿瘤辐射敏感性及癌症风险的预测指标。该程序最多可能需要5天才能完成。