ABSTRACT

Glioblastoma (GBM) is the most frequent tumor in the central nervous system. Long non-coding RNAs (lncRNAs) have been widely accepted as essential participators in cancer progression. Nonetheless, the specific role and mechanism of lncRNA SRY-box transcription factor 2 overlapping transcript (SOX2-OT) in GBM have not been studied. We evaluated expression levels of SOX2-OT, miR-192-5p and Ras-related protein Rab-2A (RAB2A) in GBM cells via qRT-PCR. To investigate the roles of SOX2-OT in GBM cells, CCK-8, JC-1, EdU, and western blot assays were performed. The connection among SOX2-OT, miR-192-5p and RAB2A in GBM cells was explored through pull down, luciferase reporter, and RIP assays. Western blot and qRT-PCR were employed to analyze the activity of extracellular-signal-regulated kinase (ERK) signaling pathway. SOX2-OT expression was higher in GBM cell lines than in normal cells. SOX2-OT knockdown repressed proliferation and promoted apoptosis of GBM cells. Mechanism assays revealed that SOX2-OT could sponge miR-192-5p. Moreover, RAB2A was certified to be the target gene of miR-192-5p. Overexpression of RAB2A reversed the repressive function of SOX2-OT knockdown on GBM cell growth. Furthermore, SOX2-OT activated ERK signaling pathway in GBM cells. SOX2-OT regulated miR-192-5p/RAB2A axis and ERK pathway to promote GBM cell growth.

摘要

胶质母细胞瘤 (GBM) 是中枢神经系统中最常见的肿瘤。长链非编码 RNA (lncRNA) 已被广泛接受为癌症进展的重要参与者。尽管如此，lncRNA SRY-box转录因子2重叠转录物（SOX2-OT）在GBM中的具体作用和机制尚未得到研究。我们通过 qRT-PCR 评估了 GBM 细胞中 SOX2-OT、miR-192-5p 和 Ras 相关蛋白 Rab-2A (RAB2A) 的表达水平。为了研究 SOX2-OT 在 GBM 细胞中的作用，进行了 CCK-8、JC-1、EdU 和蛋白质印迹分析。通过下拉、荧光素酶报告基因和 RIP 分析探索了 GBM 细胞中 SOX2-OT、miR-192-5p 和 RAB2A 之间的联系。Western印迹和qRT-PCR用于分析细胞外信号调节激酶（ERK）信号通路的活性。GBM 细胞系中的 SOX2-OT 表达高于正常细胞。SOX2-OT 敲低抑制增殖并促进 GBM 细胞的凋亡。机制分析表明，SOX2-OT 可以海绵 miR-192-5p。此外，RAB2A被证明是miR-192-5p的靶基因。RAB2A 的过表达逆转了 SOX2-OT 敲低对 GBM 细胞生长的抑制功能。此外，SOX2-OT 激活 GBM 细胞中的 ERK 信号通路。SOX2-OT 调节 miR-192-5p/RAB2A 轴和 ERK 通路促进 GBM 细胞生长。RAB2A 的过表达逆转了 SOX2-OT 敲低对 GBM 细胞生长的抑制功能。此外，SOX2-OT 激活 GBM 细胞中的 ERK 信号通路。SOX2-OT 调节 miR-192-5p/RAB2A 轴和 ERK 通路促进 GBM 细胞生长。RAB2A 的过表达逆转了 SOX2-OT 敲低对 GBM 细胞生长的抑制功能。此外，SOX2-OT 激活 GBM 细胞中的 ERK 信号通路。SOX2-OT 调节 miR-192-5p/RAB2A 轴和 ERK 通路促进 GBM 细胞生长。