miR-20a通过调控自噬网络影响子宫内膜异位症发病的机制研究

Endometriosis is a kind of chronic benign disease, which has similar features of malignant tumors like proliferation, invasion and angiogenesis. The pathogenesis of endometriosis is still poorly understood. It has been recently reported that miRNA play an important role in the development and progression of various diseases including malignant tumors through regulating autophagy.In the previous study, we found that miR-20a in serum of patients with endometriosis was significantly down-regulated by using high-throughput sequencing technology and Real-time PCR. PTEN、DUSP-2、ATG7、ATG16L1, genes closely related to cell autophagy, were predicted to be the potential target genes of miR-20a by bioinformatics methods. Based on our previous findings, we use Luciferase Reporter Assay to demonstrate the binding of miR-20a and target genes. We will do a series of experiments in vitro and in vivo and establish the model of autophagy to evaluate the hypothesis, which indicates that through inhibiting its target genes in the autophagy-related signaling pathway and autophagy process, miR-20a can regulate the autophagy level of endometrial cells, contributing to a decrease in proliferation, invasion, migration, angiogenesis capacity of endometrial cells, hindering the occurrence and development of endometriosis. This study will provide new insights into the mechanisms of endometriosis and lay the foundation to explore potential therapeutic targets for endometriosis.

子宫内膜异位症(EMs)是一种具有增殖、侵袭、血管形成等恶性肿瘤特点的慢性良性疾病，其发病机制至今仍无明确定论。目前研究证实miRNA通过对细胞自噬水平的调控，在恶性肿瘤等多种疾病发生、发展中起重要作用。课题组前期采用高通量miRNA测序和PCR检测方法，发现miR-20a在EMs患者血清中表达显著下调；通过生物信息学分析发现PTEN、DUSP-2、ATG7、ATG16L1与miR-20a存在着潜在的靶向调控关系，且功能与细胞自噬紧密相关。本研究拟以此为切入点，以荧光素酶报告基因技术验证以上基因是miR-20a的直接靶点；利用体内外实验及细胞自噬模型以期验证假说：miR-20a通过靶向抑制以上靶基因调控自噬相关信号通路及自噬的延伸过程，导致子宫内膜细胞自噬水平改变，进而抑制其增殖、侵袭、迁移、血管生成，阻碍EMs发生发展。此将阐明EMs发病的新分子机制，为发现新的EMs的防治靶标奠定基础。

子宫内膜异位症(EMs)是一种具有增殖、侵袭等恶性肿瘤特点的慢性疾病，其临床表现多样、发病机制至今仍无明确定论。MiR-20a参与肿瘤的侵袭、转移，但在EMs中的作用尚不清楚。本研究构建了EMs的小鼠模型、培养原代EMs间质细胞、收集各组织标本，采用RT-PCR, western blotting, CCK8和transwell等实验, 证实miR-20a通过调控细胞的自噬水平、增殖和迁移能力，参与EMs的发生发展。我们的实验结果显示：MiR-20a在EMs病灶组织和间质细胞中表达均显著上调；MiR-20a抑制EMs细胞自噬和凋亡水平、促进细胞的增殖和迁移能力，同时促进小鼠内异症病灶的形成；MiR-20a通过调控自噬促进EMs细胞的增殖和迁移；生物信息学和细胞实验发现RAB22A及EZH1与miR-20a存在潜在的靶向调控关系，且参与EMs细胞的增殖和迁移。本研究利用体内外实验探究miR-20a在EMs发生发展中的作用和相关机制，为EMs的诊断、治疗提供新思路和治疗靶点。