Reproductive and Genetic Center of National Research Institute for Family Planning, Beijing, China.
Am J Obstet Gynecol. 2011 Jun;204(6):544.e9-17. doi: 10.1016/j.ajog.2011.01.045. Epub 2011 Apr 16.
The objective of the study was to investigate the expression and regulation of polycomb group (PcG) proteins in human neural tube defects (NTDs).
PcG proteins in human NTD fetuses and age-matched controls were detected by Western blot. The relation between PcG proteins and microribonucleic acids was predicted and confirmed by the bioinformatics method, real-time polymerase chain reaction (PCR), dual-luciferase activity assay, and Western blot. The trimethyl condition of histone H3 Lys27 (H3K27) was detected by immunohistochemical and immunofluorescence.
Embryonic ectoderm development protein (EED) was differentially detected in placenta, cerebral cortex, and spinal cord from NTDs and age-matched controls. MiR-30b can interact with 3'-untranslated region (UTR) of Eed and regulate endogenous EED expression in neural tissues. In addition, we found an inverse relationship between the miR-30b expression and the amount of trimethyl H3K27.
Differential expression of EED exists in the nerves system in human NTDs and that is regulated by miR-30b.
本研究旨在探讨多梳组（PcG）蛋白在人神经管缺陷（NTD）中的表达和调控。
通过 Western blot 检测人 NTD 胎盘中 PcG 蛋白的表达。采用生物信息学方法、实时聚合酶链反应（PCR）、双荧光素酶活性测定和 Western blot 预测和验证 PcG 蛋白与微小 RNA（miRNA）的关系。通过免疫组化和免疫荧光检测组蛋白 H3 赖氨酸 27（H3K27）的三甲基化状态。
在 NTD 患者的胎盘、大脑皮层和脊髓中检测到胚胎外胚层发育蛋白（EED）的差异表达。miR-30b 可与 Eed 的 3'-非翻译区（UTR）相互作用，调节神经组织中内源性 EED 的表达。此外，我们发现 miR-30b 的表达与三甲基 H3K27 的量呈负相关。
在人 NTD 中神经系统中存在 EED 的差异表达，其受 miR-30b 调控。
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