异常上调的胆碱磷脂代谢是癌症的一种新兴标志，磷脂酰胆碱生产的关键酶脯氨酸激酶α（CHKα）通过未定义的机制在许多类型的人类癌症中过度表达。在这里，我们证明了糖酵解酶烯醇酶-1（ENO1）的表达水平与人类胶质母细胞瘤标本中的CHKα表达水平呈正相关，并且通过翻译后的调节严密控制CHKα的表达水平。从机械上讲，我们揭示了ENO1和泛素E3连接酶TRIM25与CHKα相关联。肿瘤细胞中高表达的ENO1与CHKα的I199 / F200结合，从而废除了CHKα和TRIM25之间的相互作用。这种作用导致抑制K195处的TRIM25介导的CHKα的多泛素化，增加了CHKα的稳定性，增强了胶质母细胞瘤细胞的胆碱代谢，促进了脑肿瘤的生长。此外，ENO1和CHKα的表达水平与胶质母细胞瘤患者的不良预后有关。这些发现强调了ENO1在胆碱磷脂代谢中的关键活化作用，并为糖化和脂质酶之间相互作用的综合调节提供了前所未有的见解。

Aberrantly upregulated choline phospholipid metabolism is a novel emerging hallmark of cancer, and choline kinase α (CHKα), a key enzyme for phosphatidylcholine production, is overexpressed in many types of human cancer through undefined mechanisms. Here, we demonstrate that the expression levels of the glycolytic enzyme enolase-1 (ENO1) are positively correlated with CHKα expression levels in human glioblastoma specimens and that ENO1 tightly governs CHKα expression via posttranslational regulation. Mechanistically, we reveal that both ENO1 and the ubiquitin E3 ligase TRIM25 are associated with CHKα. Highly expressed ENO1 in tumor cells binds to I199/F200 of CHKα, thereby abrogating the interaction between CHKα and TRIM25. This abrogation leads to the inhibition of TRIM25-mediated polyubiquitylation of CHKα at K195, increased stability of CHKα, enhanced choline metabolism in glioblastoma cells, and accelerated brain tumor growth. In addition, the expression levels of both ENO1 and CHKα are associated with poor prognosis in glioblastoma patients. These findings highlight a critical moonlighting function of ENO1 in choline phospholipid metabolism and provide unprecedented insight into the integrated regulation of cancer metabolism by crosstalk between glycolytic and lipidic enzymes.