Stathmin-1对母-胎界面滋养层细胞侵袭润性的调控作用及其分子机制研究

Gestational associated diseases are complex complication of human pregnancy, and are intractable diseases with huge financial and personal implications. The etiology of these diseases is related with impaired trophoblastic invasion. Our studies have demonstrated that stathamin-1 (STMN1) regulates the invasion ability of trophoblastic cells. This project aims to investigate the function of STMN1 in trophoblastic invasion and the underlying molecular mechanisms. Further，This project will detect these signaling pathway involved in invasion ability of trophoblastic by knockdown of STMN1 and further screen gene expression of the trophoblastic by using mRNA microarray. By using bioinformatics, animal experiment, CHIP, EMSA, western blot, immunofluorescence, flow cytometry and transwell methods, this project will confirm on the up- and down-stream molecules of STMN1 regulated trophoblastic invasion. Finally, these results will clarify the molecular mechanisms of STMN1 regulation trophoblastic invasion. In addition, this project will help to discover the biological function of STMN1, expand our understanding of the pathological mechanisms of gestational associated diseases, and provide experimental evidence for the prevention and treatment of gestational associated diseases.

妊娠相关疾病是人类怀孕期间综合症且顽固难治，给育龄妇女造成肉体和精神上的巨大痛苦。这些疾病的发生常常与滋养细胞生物学行为的改变有关。我们的研究发现一种细胞骨架调控蛋白stathmin-1 (STMN1)具有调控母-胎界面滋养细胞侵润性的作用。因此，本课题拟深入研究STMN1调控滋养细胞侵润的上下游分子机制。本项目首先检测STMN1对滋养细胞侵润重要信号通路的调控，并通过基因芯片筛查敲低STMN1对滋养细胞中侵润相关基因表达的影响。进一步运用生物信息学， western blot，细胞免疫荧光，流式细胞检测，transwell，ChIP, EMSA及动物实验等方法确认STMN1调控滋养细胞侵润的上下游分子，以阐明STMN1调控滋养层细胞侵润的分子机制。本项目的研究有助于阐明STMN1的生物学功能，拓展我们对妊娠相关疾病发生发展的认识，为妊娠相关疾病的预防和诊治，提供实验依据。

在项目的支持下，我们报道了低表达的STMN1通过调控E-cardherin/b-catein信号通路抑制细胞的增值和迁移能力，M1型巨噬细胞通过分泌TNF-α抑制STMN1蛋白的表达。我们的结果证实STMN1是一个细胞迁移和增值相关的微管调控蛋白从而在复发性流产的病理中扮演重要角色。这些结果已经发表在病理学权威杂志美国病理学上面发表。此外，我们发现在复发性流产病人的绒毛组织中YY1表达显著降低，敲低YY1显著的降低滋养层细胞的侵袭和增值能力。在绒毛外植体模型中敲低YY1可以显著的降低MMP2蛋白的表达。这些结果已经发表在病理学权威杂志病理学杂志上面发表。更进一步，我们使用RT2非编码RNA PCR芯片实验，发现改变YY1蛋白表达可以显著影响多个长链非编码RNA表达，特别是HOTAIR表达发生显著改变。我们进一步探索了YY1和HOTAIR表达改变和复发性流产之间的关联。这些结果已经发表在生物治疗权威杂志Molecular Therapy上面发表。