Gao Xiao-Zeng, Ma Ru-Hua, Zhang Zhao-Xia
Department of Anesthesiology, North China University of Science and Technology, Tangshan, China.
Emergency Department, Rizhao Hospital of Traditional Chinese Medicine, Rizhao, China.
Front Neurol. 2020 Jun 10;11:436. doi: 10.3389/fneur.2020.00436. eCollection 2020.
Ischemic stroke (IS) is a common cerebrovascular disease characterized by insufficient blood blow to the brain and the second leading cause of death as well as disability worldwide. Recent literatures have indicated that abnormal expression of miR-339 is closely related to IS. In this study, we attempted to assess the biological function of miR-339 and its underlying mechanism in IS. By accessing the GEO repository, the expression of miR-339, FGF9, and CACNG2 in middle cerebral artery occlusion (MCAO) and non-MCAO was evaluated. PC12 cells after oxygen-glucose deprivation/reoxygenation (OGD/R) treatment were prepared to mimic the IS model. The levels of miR-339, FGF9, CACNG2, and MAPK-related markers were quantitatively measured by qRT-PCR and Western blot. CCK-8 and flow cytometry analyses were performed to examine cell viability and apoptosis, respectively. IS-related potential pathways were identified using KEGG enrichment analysis and GO annotations. Bioinformatics analysis and dual-luciferase reporter assay were used to predict and verify the possible target of miR-339. Our results showed that miR-339 expression was significantly increased in MCAO and OGD/R-treated PC12 cells. Overexpression of miR-339 inhibited cell viability of PC12 cells subjected to OGD/R treatment. FGF9 and CACMG2 are direct targets of miR-339 and can reverse the aggressive effect of miR-339 on the proliferation and apoptosis of OGD/R-treated PC12 cells. Moreover, miR-339 mediated the activation of the MAPK pathway, which was inhibited by the FGF9/CACNG2 axis in PC12 cells treated by OGD/R stimulation. In summary, these findings suggested that miR-339 might act as a disruptive molecule to accelerate the IS progression via targeting the FGF9/CACNG2 axis and mediating the MAPK pathway.
缺血性中风（IS）是一种常见的脑血管疾病，其特征是脑部血液供应不足，是全球范围内第二大致死和致残原因。最近的文献表明，miR-339的异常表达与缺血性中风密切相关。在本研究中，我们试图评估miR-339的生物学功能及其在缺血性中风中的潜在机制。通过访问基因表达综合数据库（GEO），评估了大脑中动脉闭塞（MCAO）组和非MCAO组中miR-339、成纤维细胞生长因子9（FGF9）和钙通道蛋白γ2（CACNG2）的表达。制备氧糖剥夺/复氧（OGD/R）处理后的PC12细胞以模拟缺血性中风模型。通过qRT-PCR和蛋白质免疫印迹法对miR-339、FGF9、CACNG2和丝裂原活化蛋白激酶（MAPK）相关标志物的水平进行定量测定。分别进行CCK-8和流式细胞术分析以检测细胞活力和细胞凋亡。使用京都基因与基因组百科全书（KEGG）富集分析和基因本体（GO）注释来确定与缺血性中风相关的潜在通路。利用生物信息学分析和双荧光素酶报告基因检测来预测和验证miR-339可能的靶标。我们的结果表明，在MCAO组和OGD/R处理后的PC12细胞中，miR-339的表达显著增加。miR-339的过表达抑制了OGD/R处理后的PC12细胞的活力。FGF9和CACMG2是miR-339的直接靶标，并且可以逆转miR-339对OGD/R处理后的PC12细胞增殖和凋亡的促进作用。此外，miR-339介导了MAPK通路的激活，在OGD/R刺激处理的PC12细胞中，该通路被FGF9/CACNG2轴抑制。总之，这些发现表明，miR-339可能作为一种破坏分子，通过靶向FGF9/CACNG2轴并介导MAPK通路来加速缺血性中风的进展。
