【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。更多还原

【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 amelogenesisrelated 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.更多还原