NEAT1/miR-124轴维持ESCs干性在卵巢早衰细胞治疗中的作用机制研究

Stem cell therapy provides new ideas for the treatment of premature ovarian failure, but maintenance of stemness is still one of the problems that need to be solved. LncRNAs and miRNAs play an important role in maintaining the ability and pluripotency of stem cells. Our results demonstrated that two bindingsites for miR-124 on NEAT1 by bioinformatics prediction. In our pre-experiment, we found NEAT1-overexpressed and miR-124 low-expressed in mESCS. By interference NEAT1, miR-124 expression was increased. Based on the above theory and preliminary experimental basis, we speculate that NEAT1 maintains the naive state characteristics of pluripotent stem cells by regulating miR-124. In this study, the expression of NEAT1 and miR-124 were detected by interfering with the expression of NEAT1 in mouse embryonic stem cells. The binding sites and interaction between NEAT1 and miR-124 would be understood by experiments of proliferation, differentiation, cloning and tumor formation in nude micw. The effect of NEAT1 on maintenance of stem cells revealed the interaction between NEAT1/miR-124 and the regulation mechanism of NEAT1/miR-124 axis in the maintenance of naive state characteristics of pluripotent stem cells.

干细胞治疗为卵巢早衰提供了治疗新思路，但干细胞干性维持依然是目前需要解决的问题之一。LncRNAs与miRNAs在维持干细胞自我更新能力和多能性方面均具有重要作用，我们前期在生物信息学预测发时现NEAT1上有两个miR-124结合位点，小鼠胚胎干细胞(mESCs)中NEAT1高表达而miR-124低表达，干扰NEAT1导致miR-124下游靶蛋白表达降低。基于此，我们推测NEAT1通过调控miR-124影响多能干细胞的干性。本课题拟通过干扰小鼠胚胎干细胞NEAT1表达，检测miR-124下游蛋白改变，通过增殖、分化、克隆形成及小鼠体内畸胎瘤实验等，探讨NEAT1与miR-124的靶向结合及相互作用、NEAT1对干细胞自我更新及增殖分化的影响，揭示NEAT1/miR-124之间的相互作用及NEAT1/miR-124轴在多能干细胞干性维持中的调控机制，为多能干细胞干性维持研究提供新思路。

干细胞治疗为卵巢早衰提供了治疗新思路，但干细胞干性维持依然是目前需要解决的问题之一。LncRNAs与miRNAs在维持干细胞自我更新能力和多能性方面均具有重要作用。在本研究中，我们发现：①miR-124可抑制mESC细胞增殖和促进其凋亡，干扰小鼠胚胎干细胞miR-124可使下游靶蛋白C-myc表达降低。②LNCRNA在不同来源的干细胞向HiPSCs分化过程中具有表达差异，并且其中126个LNCRNAs参与这3种不同来源的干细胞的分化过程，KEGG分析发现JAK-STAT、MARK信号通路参与LNCRNAs在不同类型的干细胞分化中的调控过程；③lncRNA-NEAT1在CC组织和细胞中的表达上调, 且lncRNA-NEAT1的表达与UCEC患者的肿瘤临床分期和远处转移呈正相关；④LncRNA-NEAT1促进HeLa和SiHa细胞的迁移和侵袭，LncRNA-NEAT1促进Ishikawa和239T细胞增殖；⑤lncRNA-NEAT1调控Ishikawa和239T细胞的EMT发生；⑥LncRNA-NEAT1促进Ishikawa和239T细胞中的CDCP1蛋白表达；⑦LncRNA-NEAT1上有两个miR-124结合位点，小鼠胚胎干细胞(mESCs)中LncRNA-NEAT1高表达而miR-124低表达，干扰LncRNA-NEAT1导致miR-124下游靶蛋白表达降低，lLncRNA-NEAT1可以在Ishikawa和239T细胞中负调节其靶标miR-124。综合上述结果证实lncRNA-NEAT通过调控miR-124可以影响多能干细胞的干性，本研究成果为多能干细胞干性维持在卵巢早衰治疗研究提供新思路。