LncRNA HCG18/miR-125b/TP53调控牙髓干细胞向神经元细胞分化的研究
LncRNA HCG18/miR-125b/TP53 regulates the differentiation of dental pulp stem cells into neurons
吴建坤 1张建 1刘学丽1
作者信息
- 1. 061000 沧州 沧州市中心医院口腔科
- 折叠
摘要
目的 旨在探讨LncRNA HCG18 通过负调控miR-125b上调TP53 对人牙髓干细胞(DPSCs)向神经元样细胞分化的影响.方法 从正畸牙拔除术中获取的前磨牙中分离人DPSCs,将DPSCs诱导分化为神经元样细胞.qRT-PCR检测分化过程不同阶段HCG18、miR-125b的表达.在DPSCs细胞中转染oe-HCG18、pc-TP53、miR-125b mimic及阴性对照,以 qRT-PCR及WB方法检测各组Nestin、β-Ⅲ tubulin、NSE的mRNA和蛋白,高尔基染色法评定各组树突棘密度以检测神经元发育情况.结果 与未经诱导的DPSCs相比,DPSCs诱导分化组细胞的Nestin在分化期间先增高后降低,NSE、β-Ⅲ tubulin及miR-125b表达水平逐渐升高,HCG18 水平下调(P<0.05).HCG18 能抑制DPSCs向神经元样细胞分化,而miR-125b mimic能部分逆转HCG18 对DPSCs神经分化的抑制,TP53 能部分逆转miR-125b mimic对DPSCs神经分化的影响.结论 HCG18 能够通过负调控miR-125b激活TP53,抑制人DPSCs向神经元样细胞分化.
Abstract
Objective The purpose of this study was to explore the effects of LncRNAHCG18 on differentiation of human dental pulp stem cells(DPSCs)into neuron cells by up-regulating TP53 via negatively regulating miR-125b.Methods Human DPSCs were isolated from premolars during orthodontic operation and differentiated into neuron cells.The expression of HCG18 and miR-125b at different stages of differentiation process was measured by qRT-PCR.oe-HCG18,pc-TP53,miR-125b mimic and negative control were transfected into dPSCs.The mRNA and protein of Nestin,β-Ⅲ tubulin and NSE of neural stem cells in each group were detected by qRT-PCR and WB.The density of dendritic spines in each group was evaluated by Golgi staining to detect the development of neurons.Results Compared with undifferentiated DPSCs,the Nestin increased first and then decreased,and the expression level of NSE,β-Ⅲ tubulin and miR-125b increased gradually while the HCG18 level was inhibited during differentiation period(P<0.05).HCG18 inhibited the differentiation of DPSCs into neuron cells,while miR-125b mimics partially reverse the inhibition effects of HCG18 on DPSCs neural differentiation,TP53 partially reversed the effects of miR-125b mimic on DPSCs neural differentiation.Conclusions LncRNA HCG18 while inhibited the differentiation of DPSCs into neurons by up-regulating TP53 via negatively regulating miR-125b.
关键词
人牙髓干细胞/牙髓神经再生/lncRNA/HCG18/神经元细胞Key words
Dental pulp stem cells/Regeneration of dental nerve/lncRNA HCG18/Neuron cell引用本文复制引用
出版年
2024
NETL
NSTL
万方数据