ROLES OF NRP-1 IN BREAST CANCER DEVELOPMENT

NRP-1 在乳腺癌发展中的作用

• 批准号: 7381086
• 负责人: SNIGDHA BANERJEE
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381086
• 关键词: ROLES; NRP; BREAST; CANCER; DEVELOPMENT

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Angiogenic switch is a prerequisite for the tumor progression and presumably for the acquisition of an invasive property. Mobilization of vascular smooth muscle cells (SMCs) is the key event for new blood vessel formation. Studies have demonstrated that PDGF, secreted by various tumor cells including breast, is an important regulator of vascular smooth muscle cells motility. Recently, we have shown that NRP-1 expressed differentially in Vascular SMCs. However, the role of NRP-1 in regulation of vascular SMCs motility has not yet been fully established. We anticipate that NRP-1 may contribute in the mobilization of vascular SMCs that could be mediated through tumor cell-derived-PDGF. To test the hypothesis, we determined if PDGF physically interacts with NRP-1 to potentiate the migration of vascular SMCs. PDGF secretion level was determined by western blot analysis in different tumor cell-derived media. To determine the stimulatory effect of PDGF, cells were grown in basal media with or without PDGF in the Boyden chamber and migration was determined by counting the Giemsa stained cells. NRP-1 expression in vascular SMCs with or without PDGF was established by RT-PCR and western blot analysis. The importance of NRP-1 in PDGF-induced migration was established by silencing of NRP-1 using shRNA. Interaction of PDGF and NRP-1 was determined by immunoprecipitation/immunobloting. The studies indicate that breast tumor cells-secreted B-forms of PDGF augments the migration and NRP-1 expression in vascular SMCs. Silencing of NRP-1 by shRNA blocks the PDGF-induced migration of SMCs. B-forms of PDGF physically interacts with NRP-1 of SMCs to exert the migratory response. NRP-1 is a major component in PDGF signaling pathway in vascular SMCs migration. Next, we will determine the down-stream signaling molecules associated with this event. In particular, we will determine the role of PDGF receptors in PDGF-NRP-1 mediated vascular SMCs migration.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。血管生成开关是肿瘤进展的先决条件，并且可能是获得侵袭性的先决条件。血管平滑肌细胞（SMC）的动员是新血管形成的关键事件。研究表明，包括乳腺在内的多种肿瘤细胞分泌的PDGF是血管平滑肌细胞运动的重要调节因子。最近，我们发现 NRP-1 在血管 SMC 中表达差异。然而，NRP-1 在调节血管 SMC 运动中的作用尚未完全确定。 我们预计 NRP-1 可能有助于通过肿瘤细胞衍生的 PDGF 介导的血管 SMC 的动员。为了检验这一假设，我们确定了 PDGF 是否与 NRP-1 发生物理相互作用以增强血管 SMC 的迁移。 PDGF 分泌水平通过不同肿瘤细胞来源的培养基中的蛋白质印迹分析来测定。为了确定 PDGF 的刺激作用，细胞在 Boyden 室中在含有或不含 PDGF 的基础培养基中生长，并通过计数吉姆萨染色的细胞来确定迁移。通过 RT-PCR 和蛋白质印迹分析确定有或没有 PDGF 的血管 SMC 中 NRP-1 的表达。 NRP-1 在 PDGF 诱导的迁移中的重要性是通过使用 shRNA 沉默 NRP-1 来确定的。 PDGF 和 NRP-1 的相互作用通过免疫沉淀/免疫印迹测定。 研究表明，乳腺肿瘤细胞分泌的 B 型 PDGF 增强了血管 SMC 中的迁移和 NRP-1 表达。 shRNA 沉默 NRP-1 可阻断 PDGF 诱导的 SMC 迁移。 B 型 PDGF 与 SMC 的 NRP-1 发生物理相互作用，从而发挥迁移反应。 NRP-1 是血管 SMC 迁移过程中 PDGF 信号通路的主要成分。接下来，我们将确定与该事件相关的下游信号分子。特别是，我们将确定 PDGF 受体在 PDGF-NRP-1 介导的血管 SMC 迁移中的作用。