Yu Aixin, Wang Yiqi, Qin Jun, Lei Junrong, Bao Wendai, Dong Zhiqiang
College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430000, China.
Hubei Clinical Research Center of Central Nervous System Repair and Functional Reconstruction, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 430000, China.
J Cancer. 2024 Jun 17;15(14):4477-4489. doi: 10.7150/jca.97357. eCollection 2024.
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. 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 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 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. was identified as a hypoxia responsive gene in GBM cells. The mRNA expression level of 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 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 on GBM progression is probably dependent on the NF-κB pathway. Hypoxia-induced 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.
多形性胶质母细胞瘤（GBM）是最常见的恶性胶质瘤形式，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对GBM进展及其下游信号通路的病理功能。在常氧和低氧的U87细胞中进行RNA测序，以鉴定低氧条件下的差异表达基因（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的mRNA表达水平。进行EdU、transwell和伤口愈合试验，以评估低氧条件下其对GBM增殖和迁移的病理功能。对过表达F3的LN229细胞与对照细胞进行RNA测序和基因集富集分析，以分析富集的信号通路。用NF-κB抑制剂PDTC处理GBM细胞，并进行细胞实验以评估PDTC对OE-F3-LN229和OE-F3-U87细胞的影响。进行蛋白质免疫印迹法以验证下游信号通路。 被鉴定为GBM细胞中的缺氧反应基因。其mRNA表达水平与胶质瘤患者的总生存期呈负相关，在IV级和GBM中显著高于低级别或其他组织学类型的胶质瘤。 的过表达增强了低氧U87和LN229细胞的增殖和迁移，而敲低则抑制了它们。在OE-F3-LN229细胞中，NF-κB信号通路被激活，磷酸化p65水平升高。PDTC处理有效地挽救了低氧条件下OE-F3-LN229细胞增殖和迁移增强的情况，表明其对GBM进展的影响可能依赖于NF-κB信号通路。低氧诱导的 通过上调磷酸化p65激活NF-κB信号通路，从而促进低氧条件下GBM细胞的增殖和迁移，这可能是GBM治疗的一个潜在靶点。
