影响多种滋养细胞侵袭异常相关疾病的新靶点：miR-181b的作用与机制研究

MiR-181b is common differentially expressed in the tissue of diseases induced by trophoblast abnormal invasion using miRNA microarray. Preliminary study indicated an inverse correlation between the expression of miR-181b and trophoblast invasion. Trophoblast cells invasion ability was impaired when miR-181b was over expressed. MiR-181b was regulated by AP-2α. S1PR1 was a potential target gene of miR-181b by using target gene prediction soft wares. S1PR1 can be combined with ligand S1P and acted MEK - ERK - MMP - 2 pathway which affected trophoblast cells invasion. Based on these data, we will use gene over-expression and miRNA sponge technology to deeply explore the mechanism of miR-181b on the invasion ability of trophoblast cells and analyze the relation between miR-181b and AP-2α. Our study will help to provide a potential intervention target for the prevention and treatment of pregnancy related diseases induced by trophoblast abnormal invasion .

miR-181b是我们采用芯片筛选发现的、普遍差异表达于滋养细胞侵袭力异常相关妊娠疾病组织中的、一条高度保守的miRNA。前期发现：miR-181b表达水平与滋养细胞侵袭力呈负相关，过表达miR-181b可致滋养细胞侵袭力下降； miR-181b受胎盘特异转录因子AP-2α调控；软件分析与实验提示S1PR1为其靶点，而文献报道S1PR1可通过与配体S1P结合活化MEK-ERK-MMP-2通路影响滋养细胞侵袭力，提示 miR-181b可能通过调控S1PR1参与滋养细胞侵袭异常。研究拟：借助临床样本进一步评估miR-181b与侵袭力关系，探索其影响因素；采用过表达、敲低和挽救实验策略，明确miR-181b调控滋养细胞侵袭的作用与机制；分析miR-181b与AP-2α具体作用关系。miR-181b影响滋养细胞侵袭的作用与机制未见报道，研究可为滋养细胞侵袭力异常相关妊娠疾病的防治提供新靶标。

miR-181b是我们采用芯片筛选发现的、普遍差异表达于滋养细胞侵袭力异常相关妊娠疾病组织中的、一条高度保守的miRNA。本研究发现：1、 miR-181b 在不同物种间高度保守，且miR-181b在不同滋养细胞侵袭异常相关疾病中的表达水平与滋养细胞侵袭力呈负相关，提示其潜在强大作用；2、体外过表达实验证实miR-181b确能影响滋养细胞的侵袭力和迁移能力，并能下调侵袭相关蛋白的活性。另外，本研究亦发现miR-181b能抑制滋养细胞增殖，并影响细滋养胞周期；3、miR-181b 通过调控 S1PR1影响滋养细胞力。本研究通过萤光素酶报告基因及蛋白试验验证了S1PR1是miR-181b的靶基因，并通过过表达和挽救试验，进一步探讨了miR-181b与 S1PR1的相互作用及可能的调控机制。研究可为滋养细胞侵袭力异常相关妊娠疾病的防治提供新靶标。我们的研究工作基本上遵照合同规定进行，按项目计划，目前，已发表文章三篇，其中SCI两篇，参与编撰妇产科专著一本，申请专利一项，培养研究生两名。