Long interspersed nucleotide element-1 hypomethylation in folate-deficient mouse embryonic stem cells
Long interspersed nucleotide element-1 hypomethylation in folate-deficient mouse embryonic stem cells
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叶酸缺乏的小鼠胚胎干细胞中长散布的核苷酸元件1低甲基化
DOI：
10.1002/jcb.24496
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发表时间：
2013-07-01
影响因子：
4
通讯作者：
Zhang, Yu
中科院分区：
文献类型：
--
作者：
Chang, Shaoyan;Wang, Li;Zhang, Yu
叶酸被认为通过甲基化调节促进健康和发育。长散布核苷酸元件-1（LINE-1）在基因组的结构和功能中起着重要的调控作用，受甲基化修饰的调控。一些研究表明，叶酸浓度与LINE-1甲基化有关。然而，LINE-1甲基化和叶酸缺乏之间的直接关联仍不清楚。为探讨叶酸缺乏是否直接导致LINE-1甲基化水平降低，并分析叶酸浓度与LINE-1甲基化水平之间的关系，用不同浓度的叶酸处理小鼠胚胎干细胞，采用荧光免疫法测定其甲基化水平，ELISA法测定同型半胱氨酸含量。在不同时间点通过基质辅助激光解吸/电离飞行时间质谱法检查LINE-1甲基化。同时，观察细胞增殖和分化。结果表明，在叶酸缺乏的条件下，细胞内叶酸含量逐渐减少，而同型半胱氨酸含量逐渐增加，二者呈负相关。与叶酸正常组相比，在第18天，叶酸不足诱导的LINE-1低甲基化水平在无叶酸组中最低，在叶酸缺乏组中为中度。LINE-1甲基化水平与叶酸含量呈正相关，与同型半胱氨酸含量呈负相关。在相应的时间点，各组小鼠胚胎干细胞的增殖和分化均无变化。我们的数据表明，叶酸缺乏影响叶酸介导的一碳代谢的稳态，导致小鼠胚胎干细胞中LINE-1甲基化降低。这项研究提供了叶酸缺乏影响早期胚胎发育的初步证据。J.细胞。114：15491558，2013。(c)2013 Wiley Periodicals，Inc.
Folate is thought to contribute to health and development by methylation regulation. Long interspersed nucleotide element-1 (LINE-1), which is regulated by methylation modification, plays an important role in sculpting the structure and function of genomes. Some studies have shown that folate concentration is related to LINE-1 methylation. However, the direct association between LINE-1 methylation and folate deficiency remains unclear. To explore whether folate deficiency directly induced LINE-1 hypomethylation and to analyze the relationship between folate concentration and the LINE-1 methylation level, mouse ESCs were treated with various concentrations of folate which was measured by chemiluminescent immunoassay, and the homocysteine content was detected by ELISA. LINE-1 methylation was examined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry at various time points. Concurrently, cell proliferation and differentiation were observed. The result showed that the intracellular folate decreases under folate-deficient condition, conversely, homocysteine content increased gradually and there was a negatively correlated between them. Folate insufficiency induced LINE-1 hypomethylation at the lowest levels in folate-free group and moderate in folate-deficient group, compared with that in the folate-normal group at day 18. Moreover, LINE-1 methylation level was positively correlated with folate content, and negatively correlated with homocysteine content. At corresponding time points, proliferation and differentiation of mouse ESCs showed no alteration in all groups. Our data indicated that folate deficiency affected the homeostasis of folate-mediated one-carbon metabolism, leading to reduced LINE-1 methylation in mouse ESCs. This study provides preliminary evidence of folate deficiency affecting early embryonic development. J. Cell. Biochem. 114: 15491558, 2013. (c) 2013 Wiley Periodicals, Inc.