Light scatter and DNA accessibility to propidium iodide of ataxia telangiectasia and fanconi anemia cells.
Light scatter and DNA accessibility to propidium iodide of ataxia telangiectasia and fanconi anemia cells.
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DOI：
10.1006/bbrc.2001.5393
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发表时间：
2001-08
影响因子：
3.1
通讯作者：
C. Djuzenova;M. Flentje
中科院分区：
文献类型：
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作者：
C. Djuzenova;M. Flentje
来自患有遗传性疾病共济失调毛细血管扩张症（AT）和范可尼贫血（FA）的个体的细胞表现出对电离辐射（AT）或DNA交联剂（FA）的超敏反应，这可能是由多种因素引起的，包括染色质结构和DNA修复的缺陷。在这项研究中，一个组合的流式细胞术技术，研究AT和FA细胞的染色质构象。外周血单个核细胞（PBMC）和皮肤成纤维细胞建立AT和FA患者的细胞核进行了分析，通过光散射和荧光滴定与DNA嵌入染料碘化丙啶。光散射测量结果显示AT和FA患者的PBMC和成纤维细胞中存在粒度减小的小尺寸细胞核。荧光滴定数据可以通过假设两类具有不同亲和力的碘化丙啶结合位点来解释。AT和FA成纤维细胞中的高亲和力位点的数量显著大于对照细胞（20%）。我们的研究结果表明，适用于快速评估染色质构象的细胞计数技术，并提出了可能性，以确定AT和FA载体。
Cells from individuals with genetic diseases ataxia telangiectasia (AT) and Fanconi anemia (FA) exhibit hypersensitivity to ionizing radiation (AT) or DNA cross-linking agents (FA) which may be caused by multiple factors including defects in chromatin structure and DNA repair. In this study, a combination of cytometric techniques was employed to study the chromatin conformation of AT and FA cells. Nuclei of peripheral blood mononuclear cells (PBMCs) and of skin fibroblasts established from AT and FA patients were analyzed by light scattering and fluorimetric titration with the DNA-intercalating dye propidium iodide. The light scatter measurements revealed the presence of small-sized nuclei with reduced granularity in PBMCs and fibroblasts from both AT and FA patients. The fluorometric titration data could be interpreted by assuming two classes of propidium iodide binding sites with different affinities. The number of high-affinity sites in AT and FA fibroblasts was significantly larger (by 20%) than in control cells. Our findings show the applicability of cytometric techniques for the rapid assessment of chromatin conformation and also suggest the possibility to identify AT and FA carriers.