Geng Jiong, Shao Yun, Pu Yi, Wu Yiping, Yang Zhengxiang
Department of Emergency, Nanjing Medical University Affiliated Wuxi People's Hospital, Wuxi, Jiangsu, 214023, China.
Department of Neurosurgery, Nanjing Medical University Affiliated Wuxi People's Hospital, 299 Qingyang Road, Wuxi, Jiangsu, 214023, China.
Neurochem Res. 2025 Feb 13;50(2):100. doi: 10.1007/s11064-024-04321-7.
Glioblastoma stem like cells (GSCs) are a group of cells with strong tumorigenicity that exist in glioblastoma (GBM). Wilms tumor 1-associated protein (WTAP) is thought to promote the malignant process of GBM. However, whether WTAP regulates GSCs function to mediate GBM process is still unclear. The expression levels of WTAP and CCAAT/enhancer-binding protein delta (CEBPD) were examined by qRT-PCR and western blot. GSCs stemness, proliferation, apoptosis, and migration were assessed using sphere formation assay, CCK8 assay, EdU assay, colony formation assay, flow cytometry and transwell assay. Cell glycolysis was evaluated by testing glucose consumption and lactification. The regulation of CEBPD on WTAP was confirmed by ChIP assay and dual-luciferase reporter assay. In vivo experiments were performed to explore the effect of CEBPD/WTAP on the tumorigenicity of GSCs. WTAP and CEBPD had increased expression in GBM tissues and GSCs. Silencing of WTAP suppressed GSCs stemnness, proliferation, migration, glycolysis and promoted apoptosis. CEBPD could bind to WTAP promoter region to enhance its transcription. Besides, WTAP overexpression reversed the suppressive effect of CEBPD knockdown on GSCs stemnness, growth, migration and glycolysis in vitro, as well as the reducing effect on tumorigenicity of GSCs in vivo. CEBPD/WTAP axis played a vital role in regulating GSCs function, providing a potential therapy target for GBM.
胶质母细胞瘤干细胞样细胞(GSCs)是存在于胶质母细胞瘤(GBM)中的一组具有强致瘤性的细胞。肾母细胞瘤1相关蛋白(WTAP)被认为促进GBM的恶性进程。然而,WTAP是否通过调节GSCs功能来介导GBM进程仍不清楚。通过qRT-PCR和蛋白质印迹法检测WTAP和CCAAT/增强子结合蛋白δ(CEBPD)的表达水平。使用成球试验、CCK8试验、EdU试验、集落形成试验、流式细胞术和Transwell试验评估GSCs的干性、增殖、凋亡和迁移。通过检测葡萄糖消耗和乳酸生成来评估细胞糖酵解。通过染色质免疫沉淀试验(ChIP)和双荧光素酶报告基因试验证实CEBPD对WTAP的调控作用。进行体内实验以探究CEBPD/WTAP对GSCs致瘤性的影响。WTAP和CEBPD在GBM组织和GSCs中表达增加。敲低WTAP可抑制GSCs的干性、增殖、迁移、糖酵解并促进凋亡。CEBPD可与WTAP启动子区域结合以增强其转录。此外,WTAP过表达可逆转CEBPD敲低对体外GSCs干性、生长、迁移和糖酵解的抑制作用,以及对体内GSCs致瘤性的降低作用。CEBPD/WTAP轴在调节GSCs功能中起关键作用,为GBM提供了一个潜在的治疗靶点。