替莫唑胺在神經膠質母細胞瘤化療中得到廣泛應用，其作用機制尚不完全清楚。本文旨在探討涉及LINC00470的替莫唑胺治療神經膠質母細胞瘤的潛在分子機制。通過生物信息學分析，預測了LINC00470在神經膠質母細胞瘤中的潛在作用機制，並通過雙荧光素酶报告基因、RIP、ChIP和RNA pull-down實驗進行驗證。從神經膠質母細胞瘤患者收集的腦組織中檢測到LINC00470表達和預測的下游轉錄因子早期生長反應2（EGR2）。在神經膠質母細胞瘤細胞中進行替莫唑胺處理和/或增強和失去功能方法，測量細胞存活率、侵襲、遷移、周期分佈、血管生成、自噬和凋亡。此外，通過西方墨點法評估間葉表面標記蛋白的表達。在裸鼠中進行腫瘤移植以進行體內驗證。機制分析和生物信息學分析顯示，LINC00470透過轉錄因子EGR2轉錄活化SRY相關高運動性群組盒4（SOX4）。LINC00470和EGR2在神經膠質母細胞瘤患者的腦組織中高度表達。隨著替莫唑胺濃度的增加，LINC00470和EGR2 mRNA表達逐漸下降，替莫唑胺和LINC00470 shRNA抑制也降低了SOX4的表達。替莫唑胺治療抑制了神經膠質母細胞瘤的生長和血管生成，並增加了細胞凋亡和自噬反應，過度表達LINC00470或SOX4則抵消了這一趨勢，LINC00470 shRNA加重了這一趨勢。總之，替莫唑胺通過抑制LINC00470/EGR2/SOX4軸抑制了神經膠質母細胞瘤的進展。©2023年约翰威利和儿子有限公司出版的CNS神经科学和治疗学。

Temozolomide is extensively applied in chemotherapy for glioblastoma with unclear exact action mechanisms. This article seeks to address the potential molecular mechanisms in temozolomide therapy for glioblastoma involving LINC00470.Bioinformatics analysis was conducted to predict the potential mechanism of LINC00470 in glioblastoma, which was validated by dual-luciferase reporter, RIP, ChIP, and RNA pull-down assays. LINC00470 expression and the predicted downstream transcription factor early growth response 2 (EGR2) were detected in the collected brain tissues from glioblastoma patients. Following temozolomide treatment and/or gain- and loss-of-function approaches in glioblastoma cells, cell viability, invasion, migration, cycle distribution, angiogenesis, autophagy, and apoptosis were measured. In addition, the expression of mesenchymal surface marker proteins was assessed by western blot. Tumor xenograft in nude mice was conducted for in vivo validation.Mechanistic analysis and bioinformatics analysis revealed that LINC00470 transcriptionally activated SRY-related high-mobility-group box 4 (SOX4) through the transcription factor EGR2. LINC00470 and EGR2 were highly expressed in brain tissues of glioblastoma patients. LINC00470 and EGR2 mRNA expression gradually decreased with increasing concentrations of temozolomide in glioblastoma cells, and SOX4 expression was reduced in cells by temozolomide and LINC00470 knockdown. Temozolomide treatment induced cell cycle arrest, diminished cell viability, migration, invasion, and angiogenesis, and increased apoptosis and autophagy in glioblastoma, which was counteracted by overexpressing LINC00470 or SOX4 but was further promoted by LINC00470 knockdown. Temozolomide restrained glioblastoma growth and angiogenesis in vivo, while LINC00470 or SOX4 overexpression nullified but LINC00470 knockdown further facilitated these trends.Conclusively, temozolomide repressed glioblastoma progression by repressing the LINC00470/EGR2/SOX4 axis.© 2023 The Authors. CNS Neuroscience & Therapeutics published by John Wiley & Sons Ltd.