Long non-coding RNAs (lncRNAs) play a key role in a variety of disease processes. Plasmacytoma variant translocation 1 (PVT1), a lncRNA, is known to regulate cell functions and play a key role in the pathogenesis of many malignant tumors. The function and molecular mechanisms of lncRNA-PVT1 in cerebral ischemia remain unknown. Real-time PCR (qRT-PCR) was used to detect lncRNA-PVT1 and microRNA-30c-5p (miR-30c-5p) expression in the brain tissues of mice underwent middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen–glucose deprivation/reperfusion (OGD/R)-treated mouse primary brain neurons. Gain- or loss-of-function approaches were used to manipulate PVT1, miR-30c-5p, and Rho-associated protein kinase 2 (Rock2). The mechanism of PVT1 in ischemic stroke was evaluated both in vivo and in vitro via bioinformatics analysis, CCK-8, flow cytometry, TUNEL staining, luciferase activity assay, RNA FISH, and Western blot. PVT1 was upregulated in the brain tissues of mice treated with MCAO/R and primary cerebral cortex neurons of mice treated with OGD/R. Mechanistically, PVT1 knockdown resulted in a lower infarct volume and ameliorated neurobehavior in MCAO mice. Consistent with in vivo results, PVT1 upregulation significantly decreased the viability and induced apoptosis of neurons cultured in OGD/R. Moreover, we demonstrated that PVT1 acts as a competitive endogenous RNA (ceRNA) that competes with miR-30c-5p, thereby negatively regulating its endogenous target Rock2. Overexpression of miR-30c-5p significantly promoted cell proliferation and inhibited apoptosis. Meanwhile, PVT1 was confirmed to target miR-30c-5p, thus activating Rock2 expression, which finally led to the activation of MAPK signaling. We demonstrated that PVT1, as a ceRNA of miR-30c-5p, could target and regulate the level of Rock2, which aggravates cerebral I/R injury via activation of the MAPK pathway. These findings reveal a new function of PVT1, which helps to broadly understand cerebral ischemic stroke and provide a new treatment strategy for this disease.

长链非编码 RNA (lncRNA) 在多种疾病过程中发挥着关键作用。浆细胞瘤变异易位 1 (PVT1) 是一种 lncRNA，已知可调节细胞功能并在许多恶性肿瘤的发病机制中起关键作用。lncRNA-PVT1在脑缺血中的功能和分子机制尚不清楚。采用实时荧光定量 PCR (qRT-PCR) 检测大脑中动脉闭塞/再灌注 (MCAO/R) 和氧疗小鼠脑组织中 lncRNA-PVT1 和 microRNA-30c-5p (miR-30c-5p) 的表达– 葡萄糖剥夺/再灌注 (OGD/R) 处理的小鼠原代脑神经元。使用增益或功能丧失方法来操纵 PVT1、miR-30c-5p 和 Rho 相关蛋白激酶 2 (Rock2)。通过生物信息学分析、CCK-8、流式细胞术、TUNEL 染色、荧光素酶活性测定、RNA FISH 和蛋白质印迹。PVT1 在用 MCAO/R 治疗的小鼠的脑组织和用 OGD/R 治疗的小鼠的初级大脑皮层神经元中上调。从机制上讲，PVT1 敲低导致 MCAO 小鼠的梗塞体积减少并改善了神经行为。与体内结果一致，PVT1 上调显着降低了在 OGD/R 中培养的神经元的活力并诱导了细胞凋亡。此外，我们证明 PVT1 作为一种竞争性内源性 RNA (ceRNA)，与 miR-30c-5p 竞争，从而负调控其内源性靶标 Rock2。miR-30c-5p的过表达显着促进细胞增殖并抑制细胞凋亡。同时，PVT1 被证实靶向 miR-30c-5p，从而激活 Rock2 的表达，这最终导致了 MAPK 信号的激活。我们证明 PVT1 作为 miR-30c-5p 的 ceRNA，可以靶向和调节 Rock2 的水平，从而通过激活 MAPK 通路加重脑 I/R 损伤。这些发现揭示了 PVT1 的新功能，有助于广泛了解脑缺血性卒中并为该疾病提供新的治疗策略。