LncRNA RMST-mediated miR-107 transcription promotes OGD-induced neuronal apoptosis via interacting with hnRNPK
LncRNA RMST-mediated miR-107 transcription promotes OGD-induced neuronal apoptosis via interacting with hnRNPK
复制标题
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
DOI：
10.1016/j.neuint.2019.104644
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发表时间：
2020-02-01
影响因子：
4.2
通讯作者：
Sun, Xiu-Lan
中科院分区：
文献类型：
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作者：
Cheng, Hong;Sun, Mei;Sun, Xiu-Lan
横纹肌肉瘤2相关转录物（rhabdomyosarcoma 2-associated transcript，RMST）的长链非编码RNA（longnoncodingRNA，lncRNA）沉默已被证明在体内和体外对缺血性脑损伤和神经元损伤具有保护作用。然而，其在缺血性卒中进展中的潜在机制尚未得到很好的探讨。采用实时荧光定量PCR（qRT-PCR）检测缺氧缺糖（OGD）处理后HT-22海马神经元细胞系中RMST的表达。采用Annexin V-FITC和PI染色结合流式细胞术检测CCK-8细胞活力和凋亡细胞，以确定RMST对HT-22海马神经元细胞系的促凋亡作用。通过RNA pull-down、RNA免疫沉淀（RIP）、免疫共沉淀（Co-IP）、染色质免疫沉淀（ChIP）和双荧光素酶报告基因分析，探讨RMST在OGD诱导HT-22细胞凋亡中的作用机制。结果表明，RMST在OGD处理的HT-22细胞中高度表达。改变RMST表达导致HT-22细胞增殖和凋亡的显着变化。在HT-22细胞中，RMST通过与异质性核核糖核蛋白K（hnRNPK）相互作用，间接激活p53/miR-107信号通路，发挥其促凋亡作用。总之，我们的数据表明，RMST/hnRNPK/p53/miR-107/Bcl 212轴在调节神经元凋亡中起重要作用。
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/Bcl212 axis plays an important role in regulating neuronal apoptosis.