替莫唑胺通过抑制 LINC00470 介导的转录因子 EGR2 下调 SOX4 来防止胶质母细胞瘤的进展。

Temozolomide protects against the progression of glioblastoma via SOX4 downregulation by inhibiting the LINC00470-mediated transcription factor EGR2.

机构信息

Department of Neurosurgery, The Second Xiangya Hospital of Central South University, Changsha, China.

Department of Oncology, The Second Xiangya Hospital of Central South University, Changsha, China.

出版信息

CNS Neurosci Ther. 2023 Aug;29(8):2292-2307. doi: 10.1111/cns.14181. Epub 2023 Mar 29.

DOI:10.1111/cns.14181

PMID:36987665

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10352878/

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

Conclusively, temozolomide repressed glioblastoma progression by repressing the LINC00470/EGR2/SOX4 axis.

摘要

目的

替莫唑胺广泛应用于胶质母细胞瘤的化疗，但确切作用机制尚不清楚。本文旨在探讨 LINC00470 在替莫唑胺治疗胶质母细胞瘤中的潜在分子机制。

方法

通过生物信息学分析预测 LINC00470 在胶质母细胞瘤中的潜在作用机制，并通过双荧光素酶报告基因、RIP、ChIP 和 RNA 下拉实验进行验证。收集胶质母细胞瘤患者的脑组织样本，检测 LINC00470 表达及预测的下游转录因子早期生长反应因子 2（EGR2）。在胶质母细胞瘤细胞中采用替莫唑胺处理和/或过表达和敲低手段，检测细胞活力、侵袭、迁移、细胞周期分布、血管生成、自噬和凋亡。此外，通过 Western blot 检测间充质表面标记蛋白的表达。在裸鼠体内进行肿瘤异种移植以进行体内验证。

结果

机制分析和生物信息学分析表明，LINC00470 通过转录因子 EGR2 转录激活 SRY 相关高迁移率族盒 4（SOX4）。LINC00470 和 EGR2 在胶质母细胞瘤患者的脑组织中高表达。随着替莫唑胺浓度的增加，胶质母细胞瘤细胞中 LINC00470 和 EGR2 的 mRNA 表达逐渐降低，而细胞中 SOX4 的表达则随替莫唑胺和 LINC00470 敲低而降低。替莫唑胺处理诱导细胞周期停滞，降低细胞活力、迁移、侵袭和血管生成，并增加胶质母细胞瘤的细胞凋亡和自噬，而过表达 LINC00470 或 SOX4 可拮抗这些作用，而 LINC00470 敲低则进一步促进这些作用。替莫唑胺在体内抑制胶质母细胞瘤的生长和血管生成，而过表达 LINC00470 或 SOX4 则可消除这种作用，而 LINC00470 敲低则进一步促进这种作用。