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首页> 外文期刊>Molecular and Cellular Biology >Comparison of DNA methylation patterns among mouse cell lines by restriction landmark genomic scanning.
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Comparison of DNA methylation patterns among mouse cell lines by restriction landmark genomic scanning.

机译：通过限制性地标基因组扫描比较小鼠细胞系中DNA甲基化模式。

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Restriction landmark genomic scanning (RLGS) is a novel method which enables us to simultaneously visualize a large number of loci as two-dimensional gel spots. By this method, the status of DNA methylation can efficiently be determined by monitoring the appearance or disappearance of spots by using a methylation-sensitive restriction enzyme. In the present study, using RLGS with NotI, we examined, in comparison with a brain RLGS profile, the status of DNA methylation of more than 900 loci among three types of mouse cell lines: the embryonal carcinoma cell line P19, the stable mesenchymal cell line 10T1/2, and our established neuroepithelial (EM) cell lines. We found that the relative numbers of RLGS spots which appeared were less than 3.3% of those surveyed in all cell lines examined. However, 5 to 14% of spots disappeared, the numbers increasing with an increase in the length of the culture period, and many spots were commonly lost in 10T1/2 and in three EM cell lines. Thus, for these cell lines, many more spots disappeared than appeared. However, the numbers of spots disappearing and appearing were well balanced, and the ratio in P19 cells was almost equal to that in liver cells in vivo. These RLGS experimental observations suggested that permanent cell lines such as 10T1/2 are hypermethylated and that our newly established EM cell lines are also becoming heavily methylated at common loci. On the other hand, methylation and demethylation seem to be balanced in P19 cells in a manner similar to that in in vivo liver tissue.
机译：限制性地标基因组扫描（RLGS）是一种新颖的方法，使我们能够同时将大量基因座可视化为二维凝胶点。通过这种方法，可以通过使用甲基化敏感的限制性内切酶监测斑点的出现或消失来有效地确定DNA甲基化的状态。在本研究中，我们将RLGS与NotI结合使用，与大脑RLGS谱相比，我们检查了三种类型的小鼠细胞系中的900个基因座的DNA甲基化状态：胚胎癌细胞系P19，稳定的间充质细胞10T1 / 2系和我们建立的神经上皮（EM）细胞系。我们发现出现的RLGS斑点的相对数量少于在所有检查的细胞系中所调查的RLGS斑点的相对数量的3.3％。但是，有5％到14％的斑点消失了，数目随着培养时间的延长而增加，在10T1 / 2和3个EM细胞系中许多斑点通常丢失了。因此，对于这些细胞系，消失的斑点多于出现的斑点。但是，斑点消失和出现的数目是平衡的，并且体内P19细胞中的比率几乎等于肝细胞中的比率。这些RLGS实验观察表明，永久性细胞系（例如10T1 / 2）被高度甲基化，而我们新建立的EM细胞系在常见基因座处也被高度甲基化。另一方面，P19细胞中的甲基化和去甲基化似乎以体内肝脏组织相似的方式达到平衡。

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