微管调控蛋白CDK5RAP2在口腔鳞状细胞癌转移中的作用及其机制

The microtubule regulatory protein CDK5RAP2 plays an important role in cell migration and invasion. However, its mechanism in oral squamous cell carcinoma (OSCC) has remained unknown. We proposed that the expression of CDK5RAP2 is regulated by Wnt/β-catenin signaling pathway. High expression of CDK5RAP2 in OSCC affects tumor cell symmetric division through regulating spindle orientation in mitosis. The high expression of CDK5RAP2 promotes the cell proliferation, migration, invasion and stemness in OSCC. We aimed to determine the expression level of CDK5RAP2 in both OSCC tissues and cell lines. Through drug treatment, knock down or overexpression, we will investigate the function of CDK5RAP2 in tumor cell proliferation, migration, invasion and stemness. Moreover, we will confirm the regulation of CDK5RAP2 expression via Wnt/β-catenin signaling pathway and reveal the molecular mechanism of CDK5RAP2 in regulating symmetric cell division in mitosis. This study will not only provide new insights in the function and molecular mechanism of CDK5RAP2 in OSCC, but also suggest novel potential target for evaluation, monitor and treatment of OSCC.

微管调控蛋白CDK5RAP2对细胞迁移和侵袭有重要的作用，但其在口腔鳞状细胞癌中的作用尚无报道。根据预实验结果，我们提出“CDK5RAP2的表达受Wnt/β-catenin信号通路调控，在口腔鳞状细胞癌中，高表达的CDK5RAP2通过影响细胞有丝分裂时期纺锤体角度调控细胞的对称分裂，促进口腔癌细胞的增殖、迁移、侵袭和干细胞活性，起到促进肿瘤发生发展作用”的科学假说。本项目拟在口腔鳞癌患者组织和细胞系中证实CDK5RAP2的表达变化；通过工具药、敲低或过表达等方法，明确CDK5RAP2对肿瘤细胞的增殖、迁移、侵袭和干细胞活性等生物学行为的作用；揭示Wnt/β-catenin信号通路对CDK5RAP2表达的调控；明确CDK5RAP2对有丝分裂对称性的调控。本研究不仅有助于阐明CDK5RAP2在口腔鳞状细胞癌中的作用及机制，也为以CDK5RAP2为靶点，评估、监测和治疗口腔鳞状细胞癌提供新思路。

口腔鳞状细胞癌是一种具有高度侵袭性和致命性的恶性肿瘤，微管调节蛋白CDK5RAP2在其中的作用和机制尚不清楚。在本研究中，我们通过数据库分析和免疫组化验证CDK5RAP2在口腔鳞癌中的表达。采用药物处理、染色质免疫沉淀、荧光素酶报告实验等方法研究Wnt信号通路对CDK5RAP2的调控。通过集落形成实验、划痕实验、transwell实验、体内肿瘤发生实验和球形成实验研究CDK5RAP2在口腔鳞癌中的生物学作用。此外，通过RNA测序和生物信息学分析，鉴定CDK5RAP2缺失细胞的差异基因表达谱。最后，通过免疫染色观察纺锤体定位。结果发现CDK5RAP2在口腔鳞癌中高表达，且其表达与临床分期和淋巴结转移相关。CDK5RAP2的表达受Wnt信号通路的调控。CDK5RAP2的缺失抑制口腔鳞癌细胞的生长和迁移，并改变口腔鳞癌干细胞特征。同时，CDK5RAP2表达的抑制破坏了有丝分裂过程中的纺锤体定位。总的来说，这些结果表明CDK5RAP2是一个潜在的肿瘤干细胞标志物，并揭示了一种控制口腔鳞癌肿瘤干细胞群体的机制。