肝母细胞瘤（HB）是一种高度侵袭性的儿科恶性肿瘤，其中存在大量的癌干细胞（CSCs），与肿瘤复发和化疗耐药性有关。脑表达的X染色体蛋白1（BEX1）在纤毛生成、轴突再生和神经干细胞分化中起着关键作用。然而，在HB的代谢和干细胞程序中BEX1的作用尚不清楚。使用NCBI GEO的基因表达谱数据和免疫组化验证分析了人类和小鼠HB中的BEX1表达。使用海马驻留时间分析仪、超高压液相色谱-质谱（LC-MS）、流式细胞术、荧光定量逆转录聚合酶链式反应（qRT-PCR）、蛋白质印迹、球形形成实验和稀释的异种移植肿瘤形成实验来分析代谢和干细胞特征。我们的结果表明，BEX1的过表达显著增强了HB细胞的华尔堡效应。此外，糖酵解抑制大致削弱了BEX1对HB细胞生长和自我更新的影响，表明BEX1通过调节华尔堡效应促进了HB细胞的干细胞性维持。在机制上，BEX1通过下调过氧化物酶体增殖物激活受体联合gamma（PPARγ）来增强华尔堡效应。此外，丙酮酸脱羟酶激酶同工酶1（PDK1）在HB中对PPARγ介导的华尔堡效应抑制起到了必要的作用。此外，BEX1通过以PPARγ/PDK1依赖的方式增强华尔堡效应来支持HB的干细胞特性。BEX1和PDK1表达高的HB患者预后不良。BEX1通过调节华尔堡效应来促进HB细胞的干细胞性维持，其依赖于PPARγ/PDK1轴。百格列汀可以通过上调PPARγ来靶向BEX1介导的HB中的干细胞性特征。© 2023. 作者，Exclusive to Springer Nature Limited 授权。

Hepatoblastoma (HB) is a highly aggressive paediatric malignancy that exhibits a high presence of cancer stem cells (CSCs), which related to tumour recurrence and chemotherapy resistance. Brain expressed X-linked protein 1 (BEX1) plays a pivotal role in ciliogenesis, axon regeneration and differentiation of neural stem cells. However, the role of BEX1 in metabolic and stemness programs in HB remains unclear.BEX1 expression in human and mouse HB was analyzed using gene expression profile data from NCBI GEO and immunohistochemical validation. Seahorse extracellular flux analyzer, ultra-high-performance liquid-chromatography mass spectrometry (LC-MS), flow cytometry, qRT-PCR, Western Blot, sphere formation assay, and diluted xenograft tumour formation assay were used to analyze metabolic and stemness features.Our results indicated that overexpression of BEX1 significantly enhanced the Warburg effect in HB cells. Furthermore, glycolysis inhibition largely attenuated the effects of BEX1 on HB cell growth and self-renewal, suggesting that BEX1 promotes stemness maintenance of HB cells by regulating the Warburg effect. Mechanistically, BEX1 enhances Warburg effect through the downregulation of peroxisome proliferator-activated receptor-gamma (PPARγ). Furthermore, pyruvate dehydrogenase kinase isozyme 1 (PDK1) is required for PPARγ-induced inhibition of Warburg effect in HB. In addition, BEX1 supports the stemness of HB by enhancing Warburg effect in a PPARγ/PDK1 dependent manner.HB patients with high BEX1 and PDK1 expression had a poor prognosis. BEX1 promotes the stemness maintenance of HB cells via modulating the Warburg effect, which depends on PPARγ/PDK1 axis. Pioglitazone could be used to target BEX1-mediated stemness properties in HB by upregulating PPARγ.© 2023. The Author(s), under exclusive licence to Springer Nature Limited.