微小RNA-3960在过量全反式维甲酸诱导腭裂小鼠模型中的作用

The functions of miR-3960 in all-trans-retinoic acid induced cleft palate mouse model

目的探讨微小RNA-3960(miR-3960)在过量全反式维甲酸(atRA)诱导腭裂小鼠模型中的作用。方法6只ICR系受孕雌鼠随机分为对照组与实验组,每组3只。在E14.5,对实验组小鼠用atRA灌胃1次,对照组以相应体积植物油灌胃1次。建立过量atRA诱导腭裂小鼠模型,并收集腭突组织。采用qRT-PCR检测miR-3960在对照组和实验组腭突中的表达。利用miRBase分析miR-3960的序列特征。利用TargetScanMouse Prediction of microRNA targets预测miR-3960的靶基因。利用DAVID v6.8数据库,对miR-3960预测的靶基因进行生物信息学分析。结果miR-3960在实验组中表达上调。通过miRBase分析,只获得miR-3960在2个物种中的序列,并且mmu-miR-3960与hsa-miR-3960的序列一致。预测的靶基因共有320个,功能主要富集在细胞增殖、细胞分化、胚胎发育和组织发育等(P<0.05),信号通路主要富集在钙信号通路、cAMP信号通路和cGMP-PKG信号通路等(P<0.05)。结论在过量atRA诱导的腭裂小鼠中,上调的miR-3960可能是通过抑制腭突间充质细胞的增殖和分化而导致腭裂的发生。

Objective To research the functions of miR-3960 in all-trans-retinoic acid(atRA)induced cleft palate mouse model in order to provide the theoretical basis for gene therapy of cleft palate.Methods Excessive atRA induced cleft palate mouse model was established and palatine process tissues were collected.qRT-PCR was used to detect the expression of miR-3960.miRBase was used to analyse the characteristics of miR-3960 sequence.TargetScanMouse Prediction of microRNA targetswas used to predict the target genes of miR-3960.DAVID v6.8 database was used to perform bioinformatics analysis of target genes.Results miR-3960 was up-regulated in the experimental group.From the analysis of miRBase,we only got the sequences of miR-3960 in two species,and the two sequences were the same.There were 320 predicted target genes,the functions were mainly concentrated in cell proliferation,cell differentiation,embryo development and tissue development and so on(P<0.05),the signaling pathways were mainly concentrated in the calcium signaling pathway,cAMP signaling pathway and cGMP-PKG signaling pathway and so on(P<0.05).Conclusions In excessive atRA induced cleft palate mouse models,the up-regulated miR-3960 may result in cleft palate by inhibiting the proliferation and differentiation of palatal mesenchymal cells.