Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
Neurosurgical Institute, Fudan University, Shanghai 200040, China.
Genes (Basel). 2022 Oct 17;13(10):1882. doi: 10.3390/genes13101882.
: GBM astrocytes may adopt fetal astrocyte transcriptomic signatures involved in brain development and migration programs to facilitate diffuse tumor infiltration. Our previous data show that ETS variant 6 (ETV6) is highly expressed in human GBM and fetal astrocytes compared to normal mature astrocytes. We hypothesized that ETV6 played a role in GBM tumor progression. : Expression of ETV6 was first examined in two American and three Chinese tissue microarrays. The correlation between ETV6 staining intensity and patient survival was calculated, followed by validation using public databases-TCGA and REMBRANDT. The effect of ETV6 knockdown on glioma cell proliferation (EdU), viability (AnnexinV labeling), clonogenic growth (colony formation), and migration/invasion (transwell assays) in GBM cells was tested. RNA sequencing and Western blot were performed to elucidate the underlying molecular mechanisms. : ETV6 was highly expressed in GBM and associated with an unfavorable prognosis. ETV6 silencing in glioma cells led to increased apoptosis or decreased proliferation, clonogenicity, migration, and invasion. RNA-Seq-based gene expression and pathway analyses revealed that ETV6 knockdown in U251 cells led to the upregulation of genes involved in extracellular matrix organization, NF-κB signaling, TNF-mediated signaling, and the downregulation of genes in the regulation of cell motility, cell proliferation, PI3K-AKT signaling, and the Ras pathway. The downregulation of the PI3K-AKT and Ras-MAPK pathways were further validated by immunoblotting. : Our findings suggested that ETV6 was highly expressed in GBM and its high expression correlated with poor survival. ETV6 silencing decreased an aggressive in vitro phenotype probably via the PI3K-AKT and Ras-MAPK pathways. The study encourages further investigation of ETV6 as a potential therapeutic target of GBM.
胶质母细胞瘤中的星形胶质细胞可能会采用涉及脑发育和迁移程序的胎儿星形胶质细胞转录组特征，以促进肿瘤的弥漫性浸润。我们之前的数据表明，与正常成熟星形胶质细胞相比，ETV6 在人类胶质母细胞瘤和胎儿星形胶质细胞中高度表达。我们假设 ETV6 在胶质母细胞瘤肿瘤进展中发挥作用。
首先在两个美国和三个中国组织微阵列中检查了 ETV6 的表达。计算了 ETV6 染色强度与患者生存之间的相关性，随后使用公共数据库 TCGA 和 REMBRANDT 进行了验证。在胶质母细胞瘤细胞中，通过 EdU（增殖）、AnnexinV 标记（存活）、集落形成（克隆形成）和 Transwell 测定（迁移/侵袭）测试了 ETV6 敲低对肿瘤细胞增殖、活力、集落形成和迁移/侵袭的影响。进行了 RNA 测序和 Western blot 以阐明潜在的分子机制。
ETV6 在胶质母细胞瘤中高表达，并与不良预后相关。在神经胶质瘤细胞中沉默 ETV6 导致细胞凋亡增加或增殖、克隆形成、迁移和侵袭减少。基于 RNA-Seq 的基因表达和通路分析表明，在 U251 细胞中敲低 ETV6 导致参与细胞外基质组织、NF-κB 信号、TNF 介导的信号以及调节细胞运动、细胞增殖、PI3K-AKT 信号和 Ras 通路的基因上调。PI3K-AKT 和 Ras-MAPK 通路的下调进一步通过免疫印迹验证。
我们的研究结果表明，ETV6 在胶质母细胞瘤中高表达，其高表达与不良预后相关。沉默 ETV6 降低了体外侵袭表型，可能通过 PI3K-AKT 和 Ras-MAPK 通路。该研究鼓励进一步研究 ETV6 作为胶质母细胞瘤的潜在治疗靶点。
