WDR54介导转酮醇酶（TKT）启动结直肠癌代谢重编程的分子机制研究

肿瘤代谢重编程在CRC发生发展中扮演着重要角色。申请者前期工作中通过BioPlex，TCGA和GEO等数据库，发现在361个WD40蛋白家族成员中，WDR54具有较高的蛋白-蛋白相互作用指数，在肠癌组织中表达水平显著升高并显示了患者较差的预后；敲低该基因能够显著抑制CRC细胞增殖、侵袭和迁移能力；在对与WDR54蛋白相互作用的30个蛋白分析中，WDR54与磷酸戊糖途径非氧化途径的转酮醇酶有着较强的结合能力且敲低WDR54能够显著下调AKT，并能降低PPP途径下游产物---5-磷酸核糖和乳糖等产物的产生。为探索WDR54的促癌机制及与TKT之间的相互调控关系，本课题将通过分子间相互作用、coIP及EMSA等方式探索WDR54与TKT之间的分子调控机制，期望构建能够用于针对KRAS和BRAF突变的CRC发生、发展、复发转移和生存预后的生物标志，为CRC的靶向治疗和预后提供新的预测和干预靶点。

Tumor metabolic reprogramming plays an important role in the development of CRC. In preliminary work, through BioPlex, TCGA, and GEO databases, we found that, among 361 WD40 protein family members, WDR54 has a high protein-protein interaction index and is significantly elevated in CRC. IHC analysis with tissue microarray showed that patients with high WDR54 have poor prognosis. WDR54 knockdown significantly inhibits CRC cell proliferation, invasion, and migration. Among 30 proteins that interact with WDR54, TKT (transketolase), a key enzyme of the non-oxidative pathway of pentose phosphate pathway (PPP), has strong binding capacity with WDR54. We also found that WDR54 knockdown significantly down-regulates AKT and reduce the production of 5-riboribose and lactose which are downstream products of the PPP pathway. In order to explore the molecular mechanism of cancer promotion and the regulatory relationship between WDR54 and TKT, molecular interactions, coIP, and EMSA will be applied to verify the molecular regulation mechanisms between WDR54 & TKT and to find new biomarkers that target the occurrence, development, recurrence, metastasis, and prognosis of CRC with BRAF and KRAS mutations.