The long noncoding RNA (lncRNA) rhabdomyosarcoma 2-associated transcript (RMST) silencing has been demonstrated to protect against ischemic brain injury in vivo and neuron injury in vitro. However, its underlying mechanisms in the progression of ischemic stroke have not been well explored. The expression of RMST in oxygen-glucose deprivation (OGD)-treated HT-22 hippocampal neuron cell line was examined using quantitative Real-Time PCR (qRT-PCR). CCK-8 cell viability and apoptotic cell detection using Annexin V-FITC and PI staining coupled with flow cytometry were performed to determine the pro-apoptotic role of RMST in HT-22 hippocampal neuron cell line. Furthermore, RNA pull-down, RNA immunoprecipitation (RIP), coimmunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP) and dual-Luciferase reporter assays were performed to determine the mechanism of RMST in OGD-induced HT-22 cell apoptosis. In the results, RMST was highly expressed in OGD-treated HT-22 cells. Altered RMST expression led to marked changes in HT-22 cell proliferation and apoptosis. Mechanistically, RMST indirectly activated p53/miR-107 signaling pathway via interacting with heterogeneous nuclear ribonucleoprotein K (hnRNPK) and fulfilled its pro-apoptotic function in HT-22 cells. In conclusion, our data indicated that the RMST/hnRNPK/p53/miR-107/Bcl2l2 axis plays an important role in regulating neuronal apoptosis.

中文翻译：

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LncRNA RMST介导的miR-107转录通过与hnRNPK相互作用促进OGD诱导的神经元凋亡。

长期的非编码RNA（lncRNA）横纹肌肉瘤2相关转录（RMST）沉默已被证明可以防止体内缺血性脑损伤和体外神经元损伤。然而，其在缺血性中风进展中的潜在机制尚未得到很好的探索。使用定量实时PCR（qRT-PCR）检测了RMST在氧葡萄糖剥夺（OGD）处理的HT-22海马神经元细胞系中的表达。使用膜联蛋白V-FITC和PI染色结合流式细胞仪进行CCK-8细胞活力和凋亡细胞检测，以确定RMST在HT-22海马神经元细胞系中的促凋亡作用。此外，RNA下拉，RNA免疫沉淀（RIP），共免疫沉淀（Co-IP），进行了染色质免疫沉淀（ChIP）和双重荧光素酶报告基因检测，以确定RMST在OGD诱导的HT-22细胞凋亡中的机制。结果，RMST在OGD处理的HT-22细胞中高表达。RMST表达的改变导致HT-22细胞增殖和凋亡的明显变化。从机理上讲，RMST通过与异质核糖核蛋白K（hnRNPK）相互作用间接激活p53 / miR-107信号通路，并在HT-22细胞中实现其促凋亡功能。总之，我们的数据表明RMST / hnRNPK / p53 / miR-107 / Bcl2l2轴在调节神经元凋亡中起重要作用。RMST表达的改变导致HT-22细胞增殖和凋亡的明显变化。从机理上讲，RMST通过与异质核糖核蛋白K（hnRNPK）相互作用间接激活p53 / miR-107信号通路，并在HT-22细胞中实现其促凋亡功能。总之，我们的数据表明RMST / hnRNPK / p53 / miR-107 / Bcl2l2轴在调节神经元凋亡中起重要作用。RMST表达的改变导致HT-22细胞增殖和凋亡的明显变化。从机理上讲，RMST通过与异质核糖核蛋白K（hnRNPK）相互作用间接激活p53 / miR-107信号通路，并在HT-22细胞中实现其促凋亡功能。总之，我们的数据表明RMST / hnRNPK / p53 / miR-107 / Bcl2l2轴在调节神经元凋亡中起重要作用。