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: 6753645
• 负责人: SANJOY K. DAS
• 金额: $ 23.78万
• 依托单位: VANDERBILT UNIVERSITY
• 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 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6753645
• 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：(改编自申请者的摘要)胚胎发育过程 着床与子宫基质细胞蜕膜化有关，这是一个事件 这对怀孕的成功至关重要。去势化的过程是 以间质细胞生长和转化为蜕膜细胞为特征， 以及由于内复制而发生的基质细胞多倍体的形成 没有细胞分裂。细胞分裂的过程受到严格的调控。 细胞周期在两个特定的检查点，G1-S和G2-M。几个细胞 已知细胞周期蛋白(A、B、D和E型细胞周期蛋白)在以下方面发挥关键作用 在这些转变过程中，细胞以特定于阶段的方式生长。在这 在这方面，PI最近证明了Cyclin D3的上调是 与子宫蜕膜化密切相关。人们普遍认为 细胞周期的动态控制通过周期蛋白的相互作用来执行， 细胞周期蛋白依赖性蛋白激酶(CDK)和细胞周期蛋白依赖性蛋白依赖性蛋白激酶抑制剂(CKI)。的目标 这项修订后的申请重点放在细胞周期调节器上 参与子宫蜕膜化的发生和发展 植入过程。假设细胞周期调节器是 在子宫中表达的时空方式及其相互协调 互动在这些过程中扮演着关键角色。具体目标是：1) Cyclins(A、B、D2和E)、CDK的时空表达特征 (CDK1、CDK2、CDK4、CDK6)和CKI(p27、p21)在围着床期的表达 (1-8天)野生型小鼠子宫；2)研究细胞周期的作用 蜕膜化中的分子与EGF家族生长的影响 在这一过程中的因素；3)研究其调节和功能状态 着床期子宫中CDK、细胞周期蛋白和细胞周期蛋白I的表达 周期蛋白D2、D3或p27基因缺失小鼠的蜕膜化；以及4)评估 Cyclins(A、B、D2、D3和E)、CDKs(CDK1、CDK2、CDK4和CDK6)和CKI(p27 和p21)在Hoxa-10基因缺失小鼠的子宫中。调查人员将会描述 Northern和原位杂交检测基因表达及蛋白表达 免疫组织化学、免疫印迹和免疫沉淀。功能界别 Cyclin-CDK-CKI结合络合物的体外活性测定 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因子潜在作用的重要见解 在着床和蜕膜形成过程中的子宫生物学。因为这些 过程类似于伪对齐，结果也应该提供 与癌症生物学相关的基本信息。