在肝细胞癌（HCC）中，主要与Wnt/β-连环蛋白信号通路的异常激活相关。然而，在Wnt信号异常的HCC细胞中，细胞膜脂质组分的改变仍然不清楚。通过全面的脂质组学分析，我们揭示了6种不同HCC细胞系的细胞膜脂质组分，这些细胞系中Wnt/β-连环蛋白信号通路的组分发生了突变，导致它们的内源性信号活性发生差异。在不同调控的脂质中，二酰甘油（DAG）和鞘烷酰胺（ceramide）在Wnt3a处理后的HCC细胞膜下调。DAG和ceramide通过诱导洞蛋白介导的经典Wnt受体复合物内吞来增强Wnt/β-连环蛋白信号通路，而它们的耗竭则抑制了SNU475和HepG2细胞中的信号活性，同时减少了洞蛋白介导的内吞作用。此外，DAG和ceramide的耗竭显著阻碍了SNU475和HepG2细胞的增殖、肿瘤增长和体内迁移能力。该研究通过在HCC细胞中开创性地进行细胞膜脂质组学分析，展示了脂质在纠正癌症中异常信号通路和停止异常肿瘤生长方面的巨大潜力。本文受版权保护。版权所有。

Hepatocellular carcinoma (HCC) is largely associated with aberrant activation of Wnt/β-catenin signaling. Nevertheless, how membrane lipid composition is altered in HCC cells with abnormal Wnt signaling remains elusive. Here, by exploiting comprehensive lipidome profiling, we unravel the membrane lipid composition of six different HCC cell lines with mutations in components of Wnt/β-catenin signaling, leading to differences in their endogenous signaling activity. Among the differentially regulated lipids are diacylglycerol (DAG) and ceramide, which were downregulated at the membrane of HCC cells after Wnt3a treatment. DAG and ceramide enhanced Wnt/β-catenin signaling by inducing caveolin-mediated endocytosis of the canonical Wnt-receptor complex, while their depletion suppressed the signaling activity along with a reduction of caveolin-mediated endocytosis in SNU475 and HepG2 cells. Moreover, depletion of DAG and ceramide significantly impeded the proliferation, tumor growth, and in vivo migration capacity of SNU475 and HepG2 cells. This study, by pioneering plasma membrane lipidome profiling in HCC cells, exhibits the remarkable potential of lipids to correct dysregulated signaling pathways in cancer and stop abnormal tumor growth.This article is protected by copyright. All rights reserved.