增殖相关分子MCM和TOB1参与调控子宫内膜蜕膜化的分子机制研究

The endometrium-decidua transformation is a key event of uterine receptivity, which determines the success of blastocyst implantation and thus the term pregnancy outcome in women. However, the mechanisms governing the initiation and maintenance of endometrium-decidua transformation during human menstrual cycle remained largely unanswered. In this proposal, we propose to address this issue exploring the molecule basis of the cell cycle regulation and the maintenance of homeostasis via employing the in vivo gene knockout mice model and in vitro stromal cell culture model in combination with in situ hybridization, immunochemistry, qRT-PCR and RNA knockdown techniques. The main reseach plan includes: 1) spatiotemporal expression patterns of cell cycle associated molecule MCM and Tob1 during the periimplantation period in mice; 2) regulatory mechanisms of MCM and Tob1 during stromal decidualization in mice; 3）dynamic expression patterns of MCM and Tob1 in different stages of human endometrium; 4) concrete regulations of MCM and Tob1 during stromal decidualization in human; 5) the different pattern and potential role of MCM and Tob1 in human endometrial diseases. This project strives for systematic study on the physiological and pathological orchestration of endometrial-decidual transformation, and will provide new idea and theoretical foundation for infertility tretements and improvememnt of ART pregnancy outcome.

子宫内膜容受性是决定妊娠失败与否的主要原因，其关键标志是子宫内膜-蜕膜转变，但是这一过程是如何启动、维持，以及在人的生理周期中是如何被精确调控的尚不清楚。本项目拟解决“子宫内膜-蜕膜转化进程中细胞周期调节和稳态维持的分子机制是什么？”这一科学问题。利用在体基因敲除小鼠模型及体外基质细胞培养模型，结合原位杂交、免疫组化、qRT-PCR、RNA敲降等技术深入探讨：1）细胞增殖正向调节分子MCM家族和负向调节分子TOB1在小鼠子宫围植入期的动态表达模式；2）MCM家族和TOB1对小鼠子宫基质细胞蜕膜化的调节机制；3）MCM家族和TOB1在人子宫内膜不同阶段的动态表达模式；4）MCM和TOB1对人子宫基质细胞蜕膜化的调节机制；5）MCM和TOB1在人子宫内膜疾病中的表达变化及作用，力争系统研究子宫内膜-蜕膜转变中的生理和病理调节机制，为揭示不良妊娠机制及不孕不育的诊治提供理论基础。

子宫内膜蜕膜化是由激素调节、多种细胞周期蛋白参与的有序增殖过程，但这一过程调控机制尚不清楚。本研究主要探究了抗增殖蛋白TOB1与促增殖蛋白MCM6调控子宫内膜蜕膜化的分子机制。在研究中，我们首先利用转录本测序发现TOB1分子和MCM6分子在这一进程中表达差异显著；进一步通过对不同时期的子宫内膜临床标本检测，证实了随着子宫内膜蜕膜化进程，TOB1分子不断增加，而MCM6蛋白显著降低；为了探索其中的机制，我们体外构建了子宫内膜基质细胞蜕膜化细胞模型，干扰TOB1分子和过表达MCM6分子能够显著抑制子宫内膜蜕膜化；与此同时，细胞周期结果显示干扰TOB1分子和过表达MCM6分子能够显著加快G1-S转化过程，诱导周期蛋白表达，其具体通路机制目前还尚不清楚。结果表明MCM6蛋白以及 TOB1蛋白在子宫内膜蜕膜化进程中具有重要作用，为揭示不良妊娠机制及子宫内膜营养不良，子宫内膜增生（癌变）等疾病的分子学机制提供理论基础和新思路。