Jung Hyun-Gug, Kim Ajung, Kim Seung-Chan, Park Jae-Yong, Hwang Eun Mi
Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul 02792, Korea.
School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Korea.
Brain Sci. 2021 Jan 11;11(1):85. doi: 10.3390/brainsci11010085.
AEG-1, also called MTDH, has oncogenic potential in numerous cancers and is considered a multifunctional modulator because of its involvement in developmental processes and inflammatory and degenerative brain diseases. However, the role of AEG-1 in astrocytes remains unknown. This study aimed to investigate proteins directly regulated by AEG-1 by analyzing their RNA expression patterns in astrocytes transfected with scramble shRNA and AEG-1 shRNA. AEG-1 knockdown down-regulated TWIK-1 mRNA. Real-time quantitative PCR (qPCR) and immunocytochemistry assays confirmed that AEG-1 modulates TWIK-1 mRNA and protein expression. Electrophysiological experiments further revealed that AEG-1 further regulates TWIK-1-mediated potassium currents in normal astrocytes. An RNA immunoprecipitation assay to determine how AEG-1 regulates the expression of TWIK-1 revealed that AEG-1 binds directly to TWIK-1 mRNA. Furthermore, TWIK-1 mRNA stability was significantly increased upon overexpression of AEG-1 in cultured astrocytes ( < 0.01). Our findings show that AEG-1 serves as an RNA-binding protein to regulate TWIK-1 expression in normal astrocytes.
AEG-1，也称为MTDH，在多种癌症中具有致癌潜力，由于其参与发育过程以及炎症性和退行性脑疾病，被认为是一种多功能调节剂。然而，AEG-1在星形胶质细胞中的作用仍然未知。本研究旨在通过分析用乱序短发夹RNA（shRNA）和AEG-1 shRNA转染的星形胶质细胞中的RNA表达模式，来研究受AEG-1直接调控的蛋白质。敲低AEG-1可下调TWIK-1 mRNA。实时定量PCR（qPCR）和免疫细胞化学分析证实，AEG-1可调节TWIK-1 mRNA和蛋白质表达。电生理实验进一步表明，AEG-1可进一步调节正常星形胶质细胞中TWIK-1介导的钾电流。一项用于确定AEG-1如何调节TWIK-1表达的RNA免疫沉淀分析表明，AEG-1直接与TWIK-1 mRNA结合。此外，在培养的星形胶质细胞中过表达AEG-1后，TWIK-1 mRNA稳定性显著增加（<0.01）。我们的研究结果表明，AEG-1作为一种RNA结合蛋白，在正常星形胶质细胞中调节TWIK-1的表达。
