Regulation of Cadherins by MLK3

MLK3 对钙粘蛋白的调节

• 批准号: 10793060
• 负责人: Deborah N Chadee
• 金额: $ 45.15万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10793060
• 关键词: Adherens Junction; Adhesions; Biological Process; Breast; Cadherins; Carcinoma; Cell Adhesion; Cell-Cell Adhesion; Cells; Co-Immunoprecipitations; Complex; Development; E-Cadherin; Epithelial Cells; Epithelium; Experimental Designs; Gene Expression; Growth Factor Receptors; Human; Intercellular Junctions; Invaded; MAP Kinase Kinase Kinase; Malignant Neoplasms; Mammalian Cell; Matrix Metalloproteinases; Mediating; Mesenchymal; Mitogen-Activated Protein Kinases; Morphogenesis; Multiprotein Complexes; N-Cadherin; Neurofibromin 2; Ovarian; Phosphorylation; Phosphotransferases; Process; Proliferating; Protein Kinase; Proteins; Receptor Signaling; Regulation; Research; Role; Signal Pathway; Small Interfering RNA; Structure; Testing; Tissues; Transcription Repressor; beta catenin; cancer cell; cell motility; insight; knock-down; migration; mixed lineage kinase 3; neoplastic cell; novel; ovarian neoplasm; overexpression; protein complex; slug

ABSTRACT Mixed lineage kinase (MLK) 3 is a mitogen-activated protein kinase (MAPK) kinase kinase (MAP3K) that regulates multiple MAPK signaling pathways in mammalian cells. In addition, MLK3 has been demonstrated to regulate proliferation and invasion of different types of human epithelial cells (normal and cancer) including ovarian and breast. In the current study, we identified a novel function for MLK3 in cell-cell adhesion of ovarian epithelial TOV112D tumor cells. MLK3 siRNA knockdown disrupted cell-cell adhesion in TOV112D spheroids, reduced the cell junction protein, E-cadherin, and dramatically increased the level of Slug, a transcriptional repressor of E-cadherin. Furthermore, MLK3 was co-immunoprecipitated with E-cadherin in TOV112D cell lysates, which indicates that MLK3 may be associated with adherens junction protein complexes. Additionally, MLK3 overexpression induced the cleavage of E-cadherin resulting in an 80 kDa soluble form of E-cadherin, which has been found to activate growth factor receptor signaling. In TOV112D cells, MLK3 overexpression also strongly induced the expression of N-cadherin, a cell junction protein that is normally expressed in mesenchymal cells. Our central hypothesis is that MLK3 regulates E-cadherin and N-cadherin through multiple mechanisms, and this regulation is essential for proper cell adhesion and migration in ovarian epithelial cells. The proposed experiments are designed to define the mechanism(s) by which MLK3 regulates E- and N- cadherin, and the impact of this regulation on ovarian cell spheroid adhesion and migration. Three specific aims are proposed to test the hypothesis. In Aim 1, we will investigate MLK3 functions in E-cadherin multi- protein complexes and ovarian spheroid cell-cell adhesion. In aim 2, we will examine MLK3-dependent regulation of Slug and E-cadherin expression. In Aim 3, we will evaluate MLK3-dependent regulation of E- Cadherin cleavage, N-Cadherin expression and ovarian spheroid structure. This research will reveal novel insight into the mechanisms by which MLK3 functions to regulate E- and N-cadherin and modulate cell-cell adhesion and migration, processes which are essential for development, tissue morphogenesis, migration and cancer cell invasion.

摘要 混合谱系激酶(MLK)3是一种有丝分裂原激活的蛋白激酶(MAPK)， 在哺乳动物细胞中调节多种MAPK信号通路。此外，MLK3已经被证明 调节不同类型的人上皮细胞(正常和癌症)的增殖和侵袭，包括 卵巢和乳房。在本研究中，我们发现了MLK3在卵巢细胞间黏附中的新功能。 上皮性TOV112D肿瘤细胞。MLK3 siRNA敲除TOV112D球体中的细胞-细胞黏附， 减少细胞连接蛋白，E-钙粘素，并显著提高Slug的水平，转录 E-钙粘附素的抑制因子。此外，MLK3与E-钙粘蛋白在TOV112D细胞中免疫共沉淀 裂解产物，表明MLK3可能与黏附连接蛋白复合体有关。另外， MLK3过表达诱导E-钙粘蛋白的裂解，导致80 kDa的可溶性E-钙粘蛋白， 已经发现它可以激活生长因子受体信号。在TOV112D细胞中，MLK3过表达 也强烈诱导N-钙粘蛋白的表达，这是一种细胞连接蛋白，通常在 间充质细胞。我们的中心假设是MLK3通过多种途径调节E-钙粘蛋白和N-钙粘蛋白 这种调节对于卵巢上皮细胞的正常黏附和迁移是必不可少的。 拟议的实验旨在确定MLK3调节E-和N-的机制(S)。 钙粘附素，以及这一调节对卵巢细胞球体黏附和迁移的影响。三个具体的 为了检验这一假说，本文提出了目标。在目标1中，我们将研究MLK3在E-钙粘附素多途径中的作用。 蛋白质复合体与卵巢球状细胞-细胞黏附。在目标2中，我们将研究MLK3依赖 鼻涕蛋白和E-钙粘附素的表达调控。在目标3中，我们将评估MLK3依赖的E- 钙粘附素切割、N-钙粘附素表达与卵巢球体结构。这项研究将揭示小说 MLK3调节E-和N-钙粘蛋白及调节细胞-细胞机制的研究 黏附和迁移，对发育、组织形态形成、迁移和 癌细胞的侵袭。