Asymmetry between sister cells in a cancer cell line revealed by chemical cytometry
Asymmetry between sister cells in a cancer cell line revealed by chemical cytometry
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DOI：
10.1021/ac0495900
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发表时间：
2004-07-01
影响因子：
7.4
通讯作者：
Krylov, SN
中科院分区：
文献类型：
--
作者：
Hu, K;Ahmadzadeh, H;Krylov, SN
我们介绍了仪器和方法的双通道化学细胞仪的姐妹细胞，两个细胞出生的分裂相同的母细胞。该方法是基于毛细管电泳与激光诱导荧光检测，并允许同时探测多个胞内组分在姐妹细胞。为了测试新技术，我们比较了稳定转染GFP表达构建体的培养癌细胞中姐妹篇之间的绿色荧光蛋白（GFP）表达模式。我们发现，所有姐妹细胞在GFP表达模式中具有可检测的不对称性，置信水平高于95%。据我们所知，这是第一次报道观察到的不对称模式的蛋白质表达在姐妹细胞中的癌细胞系。该技术可以可靠地检测姐妹细胞之间化学成分的微小差异，这使得它成为研究发育过程分子机制的潜在不可或缺的工具。它将在具有复杂增殖动力学的细胞的定量研究中特别有价值（例如，干细胞）。
We introduce instrumentation and methodology for two-channel chemical cytometry of sister cells-two cells born from division of the same mother cell. The method is based on capillary electrophoresis with laser-induced fluorescence detection and allows simultaneously probing multiple intracellular components in sister cells. To test the new technology, we compared the expression patterns of green fluorescent protein (GFP) between the sisters in cultured cancer cells stably transfected with a GFP-expressing construct. We found that all sister cells had detectable asymmetry in the GFP expression patterns with a confidence level of higher than 95%. To our best knowledge, this is the first reported observation of asymmetric patterns of protein expression in sister cells in a cancer cell line. The proposed technology can reliably detect minor differences in chemical contents between sister cells, which makes it a potentially indispensable tool in studying the molecular mechanisms of developmental processes. It will be especially valuable in quantitative studies of cells with complex proliferation kinetics (e.g., stem cells).