TCF4 Promotes Neuroblastoma Proliferation and Inhibits Ferroptosis by Transactivating GPX4 Expression

Neuroblastoma, an aggressive pediatric malignancy, exhibits aberrant expression of transcription factors implicated in tumor progression. Here, we investigated the functional role of transcription factor 4 (TCF4) in neuroblastoma, focusing on its impact on cellular proliferation and ferroptosis-a regulated form of iron-dependent cell death, and elucidated the underlying molecular mechanism. Firstly, the expressions of TCF4 in neuroblastoma tissues and cell lines were analyzed, and the expressions of TCF4 mRNA and protein were significantly up-regulated. Functional analysis demonstrated that sh-TCF4 could significantly proliferate neuroblastoma cells, which was measured by CCK-8 kit, EdU staining, and clone formation experiments. Concurrently, TCF4 knockdown significantly elevated ROS accumulation and lipid peroxidation levels. Besides, sh-TCF4 decreased the levels of FTH1 and increased the TFR1 expression. Mechanistically, bioinformatic analysis using the JASPAR database predicted TCF4 binding sites within GPX4 promoter, a key ferroptosis regulator. ChIP and dual-luciferase reporter assays confirmed direct TCF4 occupancy and transcriptional activation of GPX4. Rescue experiments further validated the axis, as GPX4 overexpression abrogated the anti-proliferative and pro-ferroptotic effects induced by sh-TCF4. Collectively, the findings revealed TCF4 as a critical promoter of neuroblastoma growth and ferroptosis resistance, acting through direct up-regulation of GPX4. Targeting the TCF4-GPX4 axis may offer a novel therapeutic strategy to enhance ferroptosis sensitivity in neuroblastoma, warranting further preclinical exploration.