ASPECTS OF UTERINE CELL CYCLE REGULATION IN IMPLANTATION

着床过程中子宫细胞周期调节的各个方面

• 批准号: 6498826
• 负责人: SANJOY K. DAS
• 金额: $ 11.01万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-15 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6498826
• 关键词: cell cycle; cell differentiation; cell growth regulation; cyclin dependent kinase; cyclins; decidua; embryo implantation; enzyme activity; enzyme complex; enzyme inhibitors; epidermal growth factor; flow cytometry; gene expression; immunocytochemistry; immunoprecipitation; in situ hybridization; laboratory mouse; messenger RNA; northern blottings; phosphorylation; polyploidy; protein structure function; stromal cells; tissue /cell culture; western blottings

DESCRIPTION: (Adapted from the applicant's abstract) The process of embryo implantation is associated with uterine stromal cell decidualization, an event that is critical to the success of pregnancy. The decidualization process is characterized by stromal cell growth and transformation into decidual cells, and formation of stromal cell polyploidy which occurs due to endoreduplication without cell division. The process of cellular division is tightly regulated in the cell cycle at two particular checkpoints, G1-S and G2-M. Several cell cyclins (A, B, D and E type cyclins) are known to play key roles with regard to cellular growth in a phase specific manner during these transitions. In this regard, the PI has recently demonstrated that the upregulation of cyclin D3 is tightly associated with decidualization of the uterus. It is well accepted that the dynamic control of the cell cycle is executed by an interplay of cyclins, cyclin-dependent kinases (cdks) and cdk inhibitors (CKIs). The objectives of this revised application are to focus on the cell cycle regulators that participate in the onset and progression of decidualization aspects of the implantation process. The hypothesis is that cell cycle regulators are expressed in the uterus in a spatiotemporal manner and their coordinated interactions play key roles in these processes. The specific aims are: 1) To characterize the spatiotemporal expression of cyclins (A, B, D2 and E), cdks (cdk1, cdk2, cdk4 and cdk6) and CKIs (p27 and p21) in the periimplantation (days 1-8) uterus of wild-type mice; 2) To study the role of cell cycle molecules in decidualization and the effects of the EGF family of growth factors in this process; 3) To study the regulation and functional status of cdks, respective cyclins and CKIs in the uterus during implantation and decidualization in cyclin D2, D3 or p27 null mice; and 4) To assess the status of cyclins (A, B, D2, D3 and E), cdks (cdk1, cdk2, cdk4 and cdk6) and CKIs (p27 and p21) in the uteri of Hoxa-10 null mice. The investigators will characterize mRNA expression by Northern and in situ hybridization and protein expression by immunohistochemistry, Western blotting and immunoprecipitation. The functional activity of cyclin-cdk-CKI associated complexes will be determined by in vitro phosphorylation activity. The results obtained from these studies will provide important insights regarding the potential roles of key cell cycle regulators in uterine biology during implantation and decidualization. Since these processes are analogous to pseudomalignancy, the results should also provide basic information relevant to cancer biology.

描述：（根据申请人的摘要改编）胚胎的过程 植入与子宫基质细胞分解有关，事件 这对于怀孕的成功至关重要。 deDializatization过程是 以基质细胞的生长和转化为decidual细胞的特征， 并形成由于内置而发生的基质细胞多倍体 没有细胞分裂。细胞分裂的过程在 在两个特定检查点G1-S和G2-M处的细胞周期。几个细胞 众所周知，细胞周期蛋白（A，B，D和E型细胞周期蛋白）在 在这些过渡期间以相位特异性方式的细胞生长。在这个 考虑到，PI最近证明了细胞周期蛋白D3的上调 与子宫的切断性紧密相关。很公认 细胞周期的动态控制由细胞周期蛋白的相互作用执行 细胞周期蛋白依赖性激酶（CDKS）和CDK抑制剂（CKIS）。目标的目标 该修订的应用是将重点放在细胞周期调节器上 参与派生和进展 植入过程。假设是细胞周期调节剂是 以时空的方式在子宫中表达 交互在这些过程中起关键作用。具体目的是：1） 表征细胞周期蛋白（a，b，d2和e）的时空表达，cdks （CDK1，CDK2，CDK4和CDK6）和CKIS（p27和p21） （第1-8天）野生型小鼠的子宫； 2）研究细胞周期的作用 分子在deidatizatization中的分子和EGF生长家族的影响 在此过程中的因素； 3）研究调节和功能状态 CDK，在植入期间子宫中各自的细胞周期蛋白和CKI 细胞周期蛋白D2，D3或P27 NULL小鼠中的分解； 4）评估状态 细胞周期蛋白（A，B，D2，D3和E），CDKS（CDK1，CDK2，CDK4和CDK6）和CKIS（P27） 和p21）在Hoxa-10无效小鼠的子宫中。调查人员将描述 北部和原位杂交和蛋白质表达的mRNA表达 免疫组织化学，蛋白质印迹和免疫沉淀。功能 Cyclin-CDK-CKI相关复合物的活性将由体外确定 磷酸化活性。从这些研究中获得的结果将提供 关于关键细胞周期调节器的潜在作用的重要见解 在植入和剖析过程中的子宫生物学中。由于这些 过程类似于伪标记，结果也应提供 与癌症生物学有关的基本信息。