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环状 POLR2A 通过上调 POU3F2 促进 SOX9 转录从而促进多形性胶质母细胞瘤细胞的增殖并抑制其凋亡。

CircPOLR2A Promotes Proliferation and Impedes Apoptosis of Glioblastoma Multiforme Cells by Up-regulating POU3F2 to Facilitate SOX9 Transcription.

机构信息

Department of Interventional Radiology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong, 510080, China.

Department of Interventional Radiology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong, 510080, China.

出版信息

Neuroscience. 2022 Nov 1;503:118-130. doi: 10.1016/j.neuroscience.2022.03.035. Epub 2022 Apr 6.

Abstract

Glioblastoma multiforme (GBM) is the most common cancer in nervous system around the world. Little advancement has been achieved in promoting prognosis of GBM patients. Circular RNAs (circRNAs) are suggested as crucial effectors in modulating GBM development. Hsa_circRNA_092437 (circPOLR2A), an up-regulated circRNA in GBM cells, has not been studied yet. In this study, RT-qPCR and western blot assays were applied to detect RNA and protein levels. Cell proliferation and apoptosis were analyzed via functional assays. Subcellular fractionation assay was carried out to determine circPOLR2A distribution in cells. Bioinformatics analysis and mechanism assays were done for detecting relationships among different factors. Rescue assays were performed to confirm validity of circPOLR2A/SOX9 axis. According to experimental results, circPOLR2A was up-regulated in GBM cells and promoted GBM cell proliferation while inhibiting GBM cell apoptosis. CircPOLR2A mainly existed in cell cytoplasm and sponged miR-2113 to positively regulate POU3F2 expression. POU3F2 activated the transcription of SOX9 through interacting with SOX9 promoter (1-500). Rescue assays validated that circPOLR2A influenced GBM cell proliferation and apoptosis via SOX9. To conclude, circPOLR2A enhanced the transcription of SOX9 through miR-2113/POU3F2 axis, thus exacerbating GBM cells growth.

摘要

多形性胶质母细胞瘤(GBM)是全球神经系统最常见的癌症。在提高 GBM 患者的预后方面几乎没有取得进展。环状 RNA(circRNA)被认为是调节 GBM 发展的关键效应因子。hsa_circRNA_092437(circPOLR2A)是一种在 GBM 细胞中上调的 circRNA,尚未进行研究。在本研究中,应用 RT-qPCR 和 Western blot 检测 RNA 和蛋白水平。通过功能测定分析细胞增殖和凋亡。进行亚细胞分离测定以确定细胞中环 POLR2A 的分布。进行生物信息学分析和机制测定以检测不同因素之间的关系。进行挽救实验以确认 circPOLR2A/SOX9 轴的有效性。根据实验结果,circPOLR2A 在 GBM 细胞中上调,并促进 GBM 细胞增殖,同时抑制 GBM 细胞凋亡。circPOLR2A 主要存在于细胞质中,并通过海绵吸附 miR-2113 来正向调节 POU3F2 表达。POU3F2 通过与 SOX9 启动子(1-500)相互作用激活 SOX9 的转录。挽救实验验证了 circPOLR2A 通过 SOX9 影响 GBM 细胞增殖和凋亡。总之,circPOLR2A 通过 miR-2113/POU3F2 轴增强 SOX9 的转录,从而加剧 GBM 细胞的生长。

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