Plk2与ArgBP2相互作用在骨肉瘤细胞迁移中的作用及其机制研究

The Polo-like kinase 2(Plk2) is a serine-threonine kinase involved in many cellular processes includig cell-cycle regulation and cell survival.The Plk2 plays a crucial role in cellular signal transduction. Using yeast two-hybrid screen, we identified a novel Plk2 binding protein, ArgBP2, which was associated with cytoskeleton organization. In our previous study we confirmed Plk2 binding to the ArgBP2 specifically by GST pull-down assay, and found Plk2 phosphorylated ArgBP2 directly by kinase assay. Furthermore,Plk2 was found to be overexpressed in many osteosarcoma cell lines and had a close relationship with osteosarcoma clinical stages. So we presumed that the interaction of Plk2 and ArgBP2 may play an important role in osteosarcoma cell migration and invasion. Our project aimed to prove that Plk2 interacts with ArgBP2 and define the ArgBP2 phosphorylation sites by Plk2. Moreover,we detected the effect of this interaction in osteosarcoma cell migration and invasion and analysed the relationship of the expression of Plk2 and ArgBP2 in osteosarcoma tissues with clinical pathological characteristics and prognosis. Taken together, our study provided a novel therapeutic target for osteosarcoma.

Plk2是丝氨酸/苏氨酸激酶，参与调节细胞周期、细胞生存等多种细胞活动，是癌细胞信号转导网络的枢纽。ArgBP2是我们用酵母双杂交技术从人胎脑文库中筛选到的Plk2新的互作蛋白，在调节细胞骨架重组中起重要作用。前期我们通过GST pull-down实验显示Plk2与ArgBP2相互作用，并且激酶实验显示Plk2直接磷酸化ArgBP2，而且Plk2在多种骨肉瘤细胞系中高表达，并与骨肉瘤临床分期密切相关，推测 Plk2与ArgBP2相互作用可能在骨肉瘤细胞迁移及侵袭中起重要作用。本课题旨在验证Plk2与ArgBP2体内的相互作用，确定Plk2磷酸化ArgBP2的位点，并检测这种相互作用对骨肉瘤细胞迁移及侵袭的影响，分析临床骨肉瘤样本中Plk2和ArgBP2的表达与骨肉瘤临床病理和预后的关系，为骨肉瘤治疗提供新的靶点。

Plk2是一类保守的丝/苏氨酸激酶，参与调节细胞周期、细胞生存等多种细胞活动，是癌细胞信号转导网络的枢纽。ArgBP2是我们用酵母双杂交技术从人胎脑文库cDNA中筛选到的Plk2新的互作蛋白，在参与调节细胞骨架重组中起重要作用。前期工作我们通过酵母交配实验和GST pull down实验显示Plk2与ArgBP2发生相互作用，激酶实验显示Plk2直接磷酸化ArgBP2，而且Plk2在多种骨肉瘤细胞系中高表达，并与骨肉瘤临床分期密切相关，推测 Plk2与ArgBP2在骨肉瘤细胞迁移及侵袭中起重要作用。本课题进一步验证Plk2与ArgBP2体内的相互作用，确定Plk2磷酸化ArgBP2的位点，并检测这种相互作用对骨肉瘤细胞迁移及侵袭的影响，分析临床骨肉瘤样本中Plk2和ArgBP2的表达与骨肉瘤临床病理和预后的关系，为骨肉瘤治疗提供新的靶点。