LncRNA PVT1 regulates ferroptosis through miR-214-mediated TFR1 and p53
LncRNA PVT1 regulates ferroptosis through miR-214-mediated TFR1 and p53
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IncRNA PVT1通过miR-214介导的TFR1和P53调节铁性下垂
DOI：
10.1016/j.lfs.2020.118305
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发表时间：
2020-11-01
期刊：
影响因子：
6.1
通讯作者：
Lu, Hong
中科院分区：
文献类型：
--
作者：
Lu, Jingjing;Xu, Feng;Lu, Hong
目的：本研究旨在体内外探讨LncRNA和miRNA在脑缺血/再灌注（I/R）铁死亡中的作用。材料与方法：采用qPCR检测急性缺血性脑卒中（AIS）患者中lncRNA PVT1和miR-214的表达。然后，我们建立了脑I/R小鼠模型和OGD/R PC12细胞模型来分析铁死亡的机制。 I/R 小鼠通过侧脑室接受 lncRNA PVT 沉默或 miR-214 过表达慢病毒治疗。通过ITC染色分析梗死面积，并通过普鲁士蓝染色、铁试剂盒、MDA试剂盒、谷胱甘肽试剂盒、GPx活性试剂盒和Western blotting（WB）检测铁死亡指标。双荧光素酶报告基因测定用于评估 miR-214 是否与 PVT1、TP53 或 TFR1 结合。 Co-IP 分析了 p53 与 SLC7A11 的相互作用。主要发现：我们发现 AIS 患者血浆中 PVT1 水平上调，miR-214 水平下调。 NIHSS 评分与 PVT1 水平呈正相关，但与 miR-214 水平呈负相关。 PVT1 沉默或 miR-214 过表达可显着减少体内梗死面积并抑制铁死亡。 miR-214 过表达显着降低了 PVT1 水平。具体来说，miR-214可以结合PVT1、TP53或TFR1的3'非翻译区(3'UTR)。 PVT1 过表达或 miR-214 沉默显着消除了 Ferrostatin-1 对铁死亡指标（TFR1 表达除外）的影响。此外，miR-214沉默抵消了PVT1敲低对铁死亡相关蛋白的影响。结论：PVT1通过miR-214介导的TFR1和TP53表达调节铁死亡。 1ncRNA PVT1/miR-214/p53 可能存在正反馈环。
Aim: The study aims to investigate the roles of LncRNA and miRNA in ferroptosis in brain ischemia/reperfusion (I/R) in vivo and in vitro.Materials and methods: qPCR assay was used to analyze lncRNA PVT1 and miR-214 expressions in acute ischemic stroke (AIS) patients. Then, we established brain I/R mice models and OGD/R PC12 cell models to analyze the mechanism of ferroptosis. I/R mice were treated by lncRNA PVT silencing or miR-214 overexpressing lentivirus via lateral ventricles. Infarct size was analyzed by ITC staining, accompanied by the detection of ferroptosis indicators through Perls'Prussian blue staining, iron kit, MDA kit, glutathione kit, GPx activities kit and Western blotting (WB). Dual luciferase reporter assay was used to assess whether miR-214 bound to PVT1, TP53 or TFR1. Co-IP analyzed the interplay of p53 with SLC7A11.Key findings: We found that the levels of PVT1 were upregulated and miR-214 levels were downregulated in plasma of AIS patients. NIHSS score was positively correlated with PVT1 levels but was negatively with miR-214 levels. PVT1 silencing or miR-214 overexpression significantly reduced infarct size and suppressed ferroptosis in vivo. miR-214 overexpression markedly decreased PVT1 levels. Specifically, miR-214 could bind to 3'untranslated region (3'UTR) of PVT1, TP53 or TFR1. PVT1 overexpression or miR-214 silencing markedly abolished the effects of Ferrostatin-1 on ferroptosis indicators except for TFR1 expression. Besides, miR-214 silencing counteracted the effects of PVT1 knockdown on the ferroptosis-related proteins.Conclusion: PVT1 regulated ferroptosis through miR-214-mediated TFR1 and TP53 expression. There was a positive feedback loop of 1ncRNA PVT1/miR-214/p53 possibly.