| 王 怡,何学佳,曾雨冰,刘 帆,裴 培,王 珊.SIRT2基因通过调节WNT通路在RA诱导的神经管畸形的初步试验研究[J].,2024,(11):2009-2014 |
| SIRT2基因通过调节WNT通路在RA诱导的神经管畸形的初步试验研究 |
| Preliminary Experimental Study of SIRT2 Gene in RA-induced Neural Tube Defects by Regulating WNT Pathway |
| 投稿时间：2023-12-23 修订日期：2024-01-18 |
| DOI：10.13241/j.cnki.pmb.2024.11.002 |
| 中文关键词: 神经管畸形 RNA测序 SIRT2敲除 WNT通路 胚胎发育 |
| 英文关键词: Neural Tube Defects RNA Sequencing SIRT2 knockdown WNT pathway Embryonic development |
| 基金项目:国家自然科学基金面上项目（82071690）；首都儿科研究所所级基金项目（CXYJ-2-21-09） |
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| 中文摘要: |
| 摘要 目的：基于敲除sirtuin2（SIRT2）基因的人胚胎肾细胞（HEK293）模型的转录组学结果，在细胞水平和动物模型中探究SIRT2基因是否通过调控WNT通路参与维甲酸（Retinoic acid，RA）诱导的神经管畸形（neural tube defects， NTDs）。方法：培养SIRT2敲除组（KO-SIRT2）和正常组（KO-Con）的HEK293细胞，提取总RNA，应用转录组测序技术（RNA-seq）对上述两组细胞进行分析，寻找差异表达基因（P<0.05,|log2FoldChange|≥1）并进行GO（Gene Ontology）以及KEGG（Kyoto Encyclopedia of Genes and Genomes）分析。RA分别诱导KO-SIRT2组和KO-Con组的HEK293细胞，应用Western Blot（WB）技术检测WNT5B表达水平。建立RA诱导的NTDs小鼠模型，应用免疫组织化学染色技术（IHC）检测孕龄10.5天（E10.5）的胚鼠脑组织Sirt2和Wnt5b蛋白水平。结果：与KO-Con组相比，KO-SIRT2组的HEK293细胞中显著改变的差异表达基因有209个，KEGG分析集中在WNT、Hippo、PI3K-Akt等信号通路。敲除SIRT2组，KO-SIRT2组的WNT5B蛋白水平升高。RA诱导HEK293细胞，KO-Con组的WNT5B蛋白水平升高。IHC结果显示，RA诱导的NTDs胎鼠脑组织Sirt2蛋白和Wnt5b蛋白水平增高。结论：SIRT2基因可以通过调节WNT通路参与RA诱导NTDs的发生。 |
| 英文摘要: |
| ABSTRACT Objective: To explore whether sirtuin2 (SIRT2) regulates retinoic acid (RA) -induced neural tube defects (NTDs) at the cellular level and in animal models through WNT pathway based on the transcriptomics results of human embryonic kidney cell (HEK293) model with SIRT2 gene knockout. Methods: SIRT2 knockdown group (KO-SIRT2) and SIRT2 control group (KO-Con) were cultured and total RNA was extracted. RNA-seq technology was used to analyze the transcriptomics of the two groups of cells. Search for differentially expressed genes (P<0.05, | log2FoldChange |≥1). GO(Gene Ontology) and KEGG（Kyoto Encyclopedia of Genes and Genomes）analysis were conducted. HEK293 cells in KO-SIRT2 group and KO-Con group were induced by RA and the level of key signaling pathway protein (WNT5B) was detected by Western Blot (WB). A mouse model of NTDs induced by RA was established and the level of Sirt2 and Wnt5b proteins of brain tissues in E10.5 fetal mice was detected by immunohistochemical staining (IHC). Results: Compared with KO-Con group, 209 differentially expressed genes (107 up-regulated and 102 down-regulated) were significantly changed in KO-SIRT2 group. Enrichment analysis involved multiple signaling pathways such as WNT, Hippo and PI3K-Akt. After SIRT2 knockout, the WNT5B protein was up-regulated in the KO-SIRT2 group. After RA induction, the level of WNT5B in KO-Con group was increased. IHC showed increased levels of Sirt2 and Wnt5b proteins in the RA-induced NTDs model. Conclusion: SIRT2 gene can affect embryonic development by regulating the translation of WNT pathway genes. |
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