背景：多形性胶质母细胞瘤（GBM）是胶质瘤最常见的恶性形式，但低氧微环境中 GBM 进展的分子机制仍不清楚。本研究旨在探讨缺氧反应基因在GBM进展中的病理功能及其下游信号通路。方法：在常氧和缺氧 U87 细胞中进行 RNA-seq，以鉴定缺氧下的差异表达基因（DEG）。根据CGGA、TCGA和Rembrandt数据库的转录信息，分析胶质瘤临床样本中缺氧反应基因F3的mRNA表达水平。进行EdU、Transwell和伤口愈合实验来评价F3在缺氧条件下对GBM增殖和迁移的病理功能。进行 RNA-seq 和基因集富集分析，以分析与对照相比过表达 F3 的 LN229 细胞中的富集途径。 GBM细胞用NF-κB抑制剂PDTC处理，并进行细胞实验来评估PDTC对OE-F3-LN229和OE-F3-U87细胞的影响。进行蛋白质印迹以验证下游途径。结果：F3被鉴定为GBM细胞中的缺氧反应基因。 F3 mRNA表达水平与胶质瘤患者的总生存率呈负相关，且在IV级和GBM中较低级别或其他组织学的胶质瘤显着升高。 F3的过表达增强了缺氧U87和LN229细胞的增殖和迁移，而敲低则抑制它们。在 OE-F3-LN229 细胞中，NF-κB 通路被激活，磷酸化 p65 水平增加。 PDTC治疗有效地挽救了缺氧条件下OE-F3-LN229细胞增殖和迁移的增强，表明F3对GBM进展的影响可能依赖于NF-κB通路。结论：缺氧诱导的F3通过上调磷酸化p65激活NF-κB通路，从而促进缺氧条件下GBM细胞的增殖和迁移，可能成为GBM治疗的潜在治疗靶点。©作者。

Background: Glioblastoma multiforme (GBM) is the most common malignant form of glioma, but the molecular mechanisms underlying the progression of GBM in hypoxic microenvironment remain elusive. This study aims to explore the pathological functions of hypoxia-responsive genes on GBM progression and its downstream signaling pathways. Methods: RNA-seq was performed in normoxic and hypoxic U87 cells to identify the differentially expressed genes (DEGs) under hypoxia. The mRNA expression levels of hypoxia-responsive gene F3 in glioma clinical samples were analyzed according to the transcriptional information from CGGA, TCGA and Rembrandt databases. EdU, transwell and wound-healing assays were conducted to evaluate the pathological functions of F3 on GBM proliferation and migration under hypoxia. RNA-seq and gene set enrichment analysis were conducted to analyze the enriched pathways in LN229 cells overexpressed F3 compared to controls. GBM cells were treated with NF-κB inhibitor PDTC, and cell experiments were performed to evaluate the effects of PDTC on OE-F3-LN229 and OE-F3-U87 cells. Western blot was performed to validate the downstream pathways. Results: F3 was identified as a hypoxia responsive gene in GBM cells. The mRNA expression level of F3 was negatively correlated with the overall survival of glioma patients, and significantly increased in grade IV and GBM than lower grade or other histology of glioma. Overexpression of F3 enhanced the proliferation and migration of hypoxic U87 and LN229 cells, while knockdown inhibited them. In OE-F3-LN229 cells, the NF-κB pathway was activated, with an increased level of phosphorylated p65. PDTC treatment effectively rescued the enhanced proliferation and migration of OE-F3-LN229 cells under hypoxia, indicating that the effect of F3 on GBM progression is probably dependent on the NF-κB pathway. Conclusion: Hypoxia-induced F3 activates NF-κB pathway through upregulation of the phosphorylated p65, thus promoting the proliferation and migration of GBM cells under hypoxia, which might be a potential therapeutic target for GBM treatment.© The author(s).