牙周膜细胞内miR-23a,30a发挥靶向调控作用的基因筛选

Screening and verification of the target genes of miR-23a and miR-30a in human periodontal ligament cells

目的:预测验证牙周膜细胞内miR-23a和miR-30a的靶基因。方法:通过计算机生物信息学软件miRanda、Target Scan和Pic Tar预测miR-23a和miR-30a可能的靶基因;继而体外模拟体内炎性微环境并干预炎症牙周膜细胞内miR-23a和miR-30a的表达水平;检测已经预测到的可能靶基因及其相关蛋白的表达情况,从而分别筛选出在牙周膜细胞中miR-23a和miR-30a发挥调控作用的靶基因。结果:生物信息学分析结果显示,CAMTA1、CNTTBIP、PPPAR、Runx2、STAT1、SATB1以及STAT5B可能是miR-23a和miR-30a的靶基因,qRT-PCR筛选结果显示,STAT5B和Runx2可能分别是miR-23a和miR-30a的靶基因之一;Western Blot结果所呈现的趋势与qRT-PCR结果一致,表明STAT5B是miR-23a的靶基因之一,Runx2是miR-30a的靶基因之一。结论:miR-23a以及miR-30a参与调控炎性微环境中的牙周膜细胞的生物学活性。在牙周膜细胞中,miR-23a和miR-30a分别与STAT5B和Runx2存在靶向调控关系。

b AIM：To screen and verify the target genes of miR-23a and miR -30a in human periodontal ligament cells （PDLCs）.METHODS：Bioinformatic approaches including miRanda,TargetScan and PicTar were used to predict the potential target genes of miR-23a and miR-30a in PDLCs.Following miR-23a and miR -30a ex-pression intervention by simulated inflammation microenvironment in vitro,the mRNA and protein expression of the predicted target genes were examined in order to screen and verify the target genes of miR -23a and miR -30a.RE-SULTS：CAMTA1,CNTTBIP,PPPAR,Runx2,STAT1,STAT5B and SATB1 were predicted as the putative target genes of miR-23a and miR-30a.Real-time PCR showed that miR-23a and miR-30a gene intervention respectively af-fected the mRNA expression of STAT5B and Runx2.Western blot showed that the protein expression of STAT5B and Runx2 was respectively influenced by miR-23a and miR-30a gene intervention.CONCLUTION：MiR-23a and miR-30a are involved in the regulation of PDLCs bioactivities in the inflammatory microenvironment.STAT5B and Runx2 are respectively the target genes of miR -23a and miR -30a.