PKCzeta与VASP相互作用调节细胞骨架和细胞运动的相关机制研究

Chemotaxis plays an important role in the process of tumor metastasis. PKCzeta is a key molecular in EGF induced breast cancer cell chemotaxis signal pathway, when we use siRNA technology depleted the expression of PKCzeta, the chemotaxis ability of the cell was impaired, and the actin polymerization was reduced，but the detail machnism how PKCzeta regulated cytoskeleton was not clear. We used co-immunoprecipitation coupled with mass spectrometry, we found VASP was one of PKCzeta binding proteins，which play important role in regulating cytoskeleton. This project based on our previous studis is disinged for the following aims:1)we will investigate the binding site between PKCzeta and VASP to verify the interaction between PKCzeta and VASP and study how PKCzeta regulated activity of VASP then regulating cytoskeleton; 2)To explore the influence of VASP on EGF induced breast cancer cell chemotaxis, to further explore the role of PKCzeta in regulating cytoskeleton and cell migration. It may also suggest a potential target or biomarker for therapy patients with tumor metastasis.

在肿瘤转移过程中趋化运动起着关键作用，申请人读博期间所在课题组已发表文章表明PKCzeta是EGF诱导的乳腺癌细胞趋化运动信号通路中的关键信号分子，采用siRNA技术降低PKCzeta表达后，可以明显抑制EGF诱导的趋化运动，细胞骨架肌动蛋白聚合能力明显减弱。但目前关于趋化运动过程中PKCzeta调控的下游细胞骨架信号通路具体机制并不清楚，我们前期研究采用免疫共沉淀偶联质谱分析的方法发现VASP是PKCzeta的一个结合蛋白，VASP在调节细胞骨架中发挥重要作用，本研究拟在前期工作基础上进一步阐明VASP与PKCzeta的结合位点，明确PKCzeta对VASP及细胞骨架的调节作用，并明确VASP在EGF诱导的乳腺癌细胞趋化运动中的作用,进一步明确PKCzeta在调节细胞骨架和细胞运动中的作用，为筛选治疗乳腺癌转移提供新的评估指标和靶点，对完善乳腺癌转移的治疗提供重要线索。