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
俞秀君 1苏俭生1
作者信息
- 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
- 折叠
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.
Abstract
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Key words
long intergenic noncoding RNA-EPS/microRNA-24-3p/bone morphogenetic protein 2/enamel development引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据