lincRNA-EPS/miR-24-3p/BMP2信号轴对牙釉质发育影响的体外实验研究  

Effect of lincRNA-EPS/miR-24-3p/BMP2 signal axis on enamel development:An experimental study in vitro

作　　者：俞秀君 苏俭生[1] YU Xiujun;SU Jiansheng(Shanghai Engineering Research Center of Tooth Restoration and Regeneration&Tongji Research Institute of Stomatology&Department of Prosthodontics,Shanghai Tongji Stomatological Hospital and Dental School,Tongji University,Shanghai 200072,China)

机构地区：[1]上海市同济口腔医院口腔修复科,同济大学口腔医学院,上海牙组织修复与再生工程技术研究中心,同济大学口腔医学研究所,上海200072

出　　处：《口腔颌面外科杂志》2024年第6期427-434,共8页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(82170913);上海市科学技术委员会项目(201409006200)。

摘　　要：目的:在体外探究长链基因间非编码RNA-EPS(long intergenic non-coding RNA-EPS,lincRNA-EPS)/微小RNA(microRNA,miRNA)-24-3p/骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)信号轴对牙釉质发育的影响。方法:将lincRNA-EPS过表达质粒和miR-24-3p mimics转染至LS8细胞中,并通过实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)分别检测其转染效率;应用RT-qPCR分别检测转染后成釉相关基因(AMELX、AMBN、AMTN、MMP20)和BMP2的mRNA相对表达水平;应用蛋白质印迹法(Western blotting)检测转染后BMP2的蛋白表达水平;采用独立样本t检验进行组间定量比较。结果:显微镜下荧光图像、RT-qPCR结果显示,lincRNA-EPS过表达质粒和miR-24-3p mimics均转染成功;lincRNA-EPS过表达质粒转染后成釉相关基因(AMELX、AMBN、AMTN、MMP20)、BMP2的mRNA及蛋白表达均显著升高,miR-24-3p mimics转染后成釉相关基因、BMP2的mRNA及蛋白表达均显著降低,lincRNA-EPS能在一定程度上抑制miR-24-3p对BMP2的调控作用,差异具有统计学意义(P<0.05)。结论:牙釉质发育受lincRNA-EPS/miR-24-3p/BMP2信号轴的调控,lincRNA-EPS可以影响miR-24-3p对下游BMP2的表达抑制,从而促进釉质发育。Objective:To investigate the effect of long intergenic non-coding RNA-EPS(lincRNA-EPS)/microRNA(miRNA)-24-3p/bone morphogenetic protein 2(BMP2)signal axis on enamel development in vitro.Methods:The overexpressed plasmid of lincRNA-EPS and miR-24-3p mimics were transfected into LS8 cells and their transfection efficiency was detected by real-time quantitative polymerase chain reaction(RT-qPCR).RT-qPCR was used to detect the relative mRNA expression of amelogenesis-related genes(AMELX,AMBN,AMTN,MMP20)and BMP2 after transfection.Western blotting was used to detect the protein expression of BMP2 after transfection.Independent sample t test was used for quantitative comparison between groups.Results:Fluorescence images under microscope and RT-qPCR results showed that lincRNA-EPS overexpression plasmid and miR-24-3p mimics were transfected successfully.The relative mRNA expression of amelogenesis-related genes and BMP2 as well as the protein expression of BMP2 were significantly increased after transfection with lincRNA-EPS overexpression plasmid,while the relative mRNA expression of amelogenesis-related genes and BMP2 as well as the protein expression of BMP2 were significantly decreased after transfection with miR-24-3p mimics.To a certain extent,lincRNA-EPS can inhibit the regulatory effect of miR-24-3p on BMP2,and the difference was statistically significant(P<0.05).Conclusion:Enamel development is regulated by lincRNA-EPS/miR-24-3p/BMP2 signal axis,and lincRNA-EPS can affect the inhibition of miR-24-3p on downstream BMP2 expression,thus promoting enamel development.

关 键 词：长链基因间非编码RNA-EPS 微小RNA-24-3p 骨形态发生蛋白2 牙釉质发育 

分 类 号：R782[医药卫生—口腔医学]