mmu-mir-221/222/500及其靶基因在叶酸缺乏小鼠神经管畸形中相关性的研究

The study on mmu-mir-221/222/500 and its target genes in folate deficiency mouse neural tube defects

目的寻找叶酸缺乏相关的神经管畸形(NTDs)候选miRNAs及其靶基因。方法在前期NTDs叶酸缺乏动物模型基础上,对已发现的3个高置信度基因组拷贝数变异(CNVs)和小鼠miRNAs基因组位置比对得到CNV覆盖下相应的miRNAs;与已建立的人类和小鼠NTD相关基因数据库比对,应用系统生物信息学分析筛选出NTDs候选miRNAs及其靶基因;并用q PCR验证miRNAs及候选靶基因。结果与已知小鼠miRNAs基因组位置比对发现8个miRNAs;进一步生物信息学分析发现,有3个NTD候选miRNAs(mmu-mir-221,mmu-mir-222,mmu-mir-500)及其12个候选靶基因;q PCR结果显示,与正常胚胎神经组织相比,NTD胚胎神经组织中9个候选基因表达显著下调(P<0.05),但在其亲代组织及胚胎其他组织中并未发现下调。结论 3个miRNAs及其9个候选靶基因与叶酸缺乏NTDs的发生密切相关。

Objective： To find candidate miRNAs of folate-deficient neural tube defects （NTDs） . Methods： Based on the folate deficiency NTD mouse model, we compared the genomic positions of known mouse miRNAs with the CNVs to find the CNV-covered miRNAs; matched these target genes with the NTD-related genes and then in-depth bioinformatics analysis to find the miRNAs and its target genes which were identified as potential susceptibility candidates in NTDs; and qPCR was used to verify the results. Results： By comparing the genomic positions of known mouse miRNAs with the CNVs, 8 miRNAs were identified. After the bioinformatics analysis, we found that there were 3 miRNAs （mmu-mir-221, mmu-mir-222, and mmu-mir-500） and 12 targets among the NTD gene list. 9 putative genes showed downregulated expression in the single NTD embryonic neural tissues compared to single normal embryonic neural tissues in our qPCR experiments. No differences were found in the parental brain tissues or the remaining embryonic tissues （except neural tissues） . Conclusion： 3 miRNAs and 9 candidate target genes were closely related with the development of folate-deficient NTDs.