子宫内膜癌存在多种分子分型，其中DNA聚合酶基因POLE突变型预后较好。本课题组基于患者样本检测到P286R和V411L热点突变。然而突变型肿瘤预后较好的机制仍知之甚少。研究发现，其突变可能会增加DNA复制应激（DRS），而化疗药亦可激发DRS。DRS可诱导WAPL蛋白依赖的染色体内聚缺陷，引发并维持有丝分裂阻滞和有丝分裂灾难。故推测，POLE突变的内膜癌患者预后较好的机制可能是POLE-P286R突变和V411L突变引起的DRS协同化疗药物诱导的DRS作用，促进细胞杀伤作用，其分子机制可能是通过增加DNA损伤以及WAPL依赖的内聚蛋白复合物功能缺陷，促进有丝分裂灾难的发生。本项目拟通过构建P286R突变和V411L突变细胞，结合WAPL基因干预，运用时差显微镜、免疫荧光共染、荧光标记的融合蛋白细胞构建等技术，研究上述机制。为解释上述临床现象并提供潜在的分子干预靶点提供理论和实验依据。

Patients with endometrial cancer have multiple molecular types, and patient with the POLE mutant has a better prognosis. Our team have detected P286R and V411L hotspot mutations in POLE gene based on patient samples. However, the mechanism of better prognosis with mutation is still poorly understood. Studies have found that mutations may increase DNA replication stress (DRS), and chemotherapeutics can also trigger DRS. While DRS can induce WAPL protein-dependent chromosomal cohesion fatigue, induced mitotic arrest and mitotic catastrophe. Therefore, it is speculated that POLE-P286R and V411L mutation may increase DRS. POLE mutation may synergy with chemotherapy to induced cell death. The molecular mechanism may be through increasing DRS and then induced WAPL-dependent cohesion fatigure to promote mitotic catastrophe. This project intends to study the above mechanism by constructing P286R mutant and V411L mutant cells, combined with WAPL gene intervention, and using techniques such as time lapse microscope, immunofluorescence co-staining, and fluorescently labeled fusion protein cell construction. This study will provide theoretical and experimental basis for explaining the above clinical phenomena and providing potential molecular intervention targets for endometrial cancer patients.