WDR63促进人根尖牙乳头干细胞成神经分化功能  

WDR63 promotes the neural differentiation potential of human apical papilla stem cells

作　　者：周家玮 范志朋[1] ZHOU Jiawei;FAN Zhipeng(Laboratory of Beijing Stomatological Hospital,School of Stomatology,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京口腔医院研究所,北京100050

出　　处：《口腔生物医学》2024年第4期193-199,共7页Oral Biomedicine

基　　金：北京市自然科学基金项目(7222075)。

摘　　要：目的:研究WD重复结构域63(WDR63)对人根尖牙乳头干细胞(SCAPs)体外成神经分化能力的作用。方法:通过实时荧光定量PCR检测未转染慢病毒SCAPs中WDR63基因在成神经分化诱导3、6、9 d的表达变化,利用慢病毒转染分别获得WDR63稳定过表达和敲低的SCAPs,通过类神经球形成实验对各组SCAPs进行神经分化诱导,对类神经球的直径进行定量分析,免疫荧光染色实验检测SCAPs中巢蛋白(Nestin)和βⅢ-微管蛋白(βⅢ-tubulin)的表达,实时荧光定量PCR实验检测神经分化相关指标βⅢ-tubulin、神经源性分化蛋白(NeuroD)、神经细胞黏附分子(NCAM)和Nestin基因的表达。结果:未转染慢病毒SCAPs中WDR63的表达量在成神经分化后的3、6、9 d逐渐增加(P<0.05);过表达WDR63后,WDR63基因(P<0.001)和蛋白表达增加,神经球直径增加(P<0.01),Nestin和βⅢ-tubulin神经球样细胞荧光强度增加(P<0.01),βⅢ-tubulin、NeuroD、NCAM和Nestin基因表达升高(P<0.05);敲低WDR63后,WDR63基因(P<0.01)和蛋白的表达降低,神经球直径降低(P<0.05),Nestin和βⅢ-tubulin神经球样细胞荧光强度减少(P<0.05),βⅢ-tubulin、NeuroD、NCAM和Nestin基因表达降低(P<0.05)。结论:WDR63可促进SCAPs的成神经分化功能。Objective:To investigate the effect of WDR63 on the differentiation ability of human stem cells from the apical papilla(SCAPs)and the expression of neural differentiation-related markers in vitro.Methods:qRT-PCR was used to detect the expression changes of WDR63 gene in untransfected lentivirus SCAPs at 3,6 and 9 days after induction of neurogenic differentiation.Using lentiviral transfection,stable over-expression and knockdown of WDR63 in SCAPs were obtained.The neural differentiation of the cells was induced through a neurosphere formation experiment,and the diameter of the neurospheres was quantitatively analyzed.The expression of Nestin andβⅢ-tubulin in SCAPs was detected by immunofluorescence staining and quantitatively analyzed.qRT-PCR was used to detect the expression of neural differentiation-related indicators such asβⅢ-tubulin,NeuroD,NCAM,and Nestin genes.Results:The expression level of WDR63 in untransfected lentivirus SCAPs gradually increased at 3,6,and 9 days after neuronal differentiation(P<0.05).Overexpression of WDR63 led to the increase of WDR63 gene(P<0.001)and protein expression,significant increase in neuronal ball diameter(P<0.01),and obvious increased in Nestin andβⅢ-tubulin-positive neural ball-like cells with significantly enhanced fluorescence intensity(P<0.01).βⅢ-tubulin,NeuroD,NCAM,and Nestin gene expression also increased(P<0.05).Conversely,knockdown of WDR63 led to decrease WDR63 gene(P<0.01)and protein expression,significant reduction in neuronal ball diameter(P<0.05),and significant decrease in Nestin andβⅢ-tubulin-positive neural ball-like cells(P<0.05)with decreasedβⅢ-tubulin,NeuroD,NCAM,and Nestin gene expression(P<0.05).Conclusions:WDR63 promotes the neural differentiation potential of SCAPs.

关 键 词：根尖牙乳头干细胞 成神经分化 WDR63 类神经球 神经分化指标 

分 类 号：R781[医药卫生—口腔医学]