LncRNA H19与miR200a构建ceRNA网络介导胰腺癌干细胞失干性的机制

LncRNA与miRNA在肿瘤干细胞中发挥重要调控作用，它们通过一种全新调控模式：竞争性内源性RNA (ceRNA)调控网络，精准调控靶基因表达，LncRNA H19与miR200a可互为ceRNA；前期研究发现miR200a通过TGFβ2信号通道调控胰腺癌干细胞（pCSCs）干性特征，而H19在pCSCs中与miR200a对应表达。在此基础上，本项目拟完成：1、通过分别调控pCSCs中H19和miR200a活性，观测其相互竞争性表达，检测其下游靶基因的表达变化，观察干性基因的表达和干性生物学特性的变化；2、研究H19与miR200a通过构建ceRNA网络对话精准靶控下游基因表达，介导pCSCs发生间质-上皮转化(MET)并失去干性的机制；3、构建由胰腺脱细胞支架、生物反应器和灌注培养液为主体的肿瘤细胞三维培养体系，为pCSCs提供模拟肿瘤微环境的研究平台。以此为胰腺癌的治疗探寻新靶点。

Long non-coding RNA(lncRNA) and microRNA can play an important role in cancer stem cells, they can accurately regulate the expression of target gene through a new regulatory model: competing endogenous RNA(ceRNA). LncRNA H19 and miR200a can be ceRNAs each other. In the previous research, we found that miR200a could regulate the stemness characteristics of pancreatic cancer stem cells (pCSCs) by TGFβ2 signaling pathway and the expression of H19 in pCSCs was corresponding with miR200a. Based on this basis, we intend to complete the following projects:1. We observe the competitive expression of H19 and miR200a by regulation the activity of of them in pCSCs respectively, then detect the expression of its downstream target genes, observe the changes of expression of stemness genes and biological stemness characteristics. 2.Study the mechanism of H19 and miR200a controlling the expression of downstream target gene precisely through ceRNA network, mediating the mesenchymal epithelial transformation (MET) and the loss of stemness of pCSCs.3.Construct the three dimensional culture system consisting of pancreatic decellularized scaffold, bioreactor and medium to provide the simulated microenvironment for pCSCs.Then explore new targets for the treatment of pancreatic cancer.