Department of Kidney Transplantation, The Affiliated Hospital of Qingdao University, Qingdao, China.
J Cell Mol Med. 2020 Apr;24(8):4707-4715. doi: 10.1111/jcmm.15142. Epub 2020 Mar 11.
Circular RNA YAP1 (circYAP1) was reported to participate in progression of gastric cancer. However, the role of circYAP1 in acute kidney injury (AKI) remains obscure. We attempted to examine the effects of circYAP1 on ischaemia/reperfusion-stimulated renal injury. AKI model was established by treating HK-2 cells in ischaemia/reperfusion (I/R) environment. CircYAP1 expression in blood of AKI patients and I/R-treated HK-2 cells was evaluated via RT-qPCR. CCK-8, flow cytometry, ELISA and ROS assay were executed to test the impact of circYAP1 on cell viability, apoptosis, inflammatory cytokines and ROS generation. Bioinformatic analysis was executed to explore miRNA targets. The relativity between circYAP1 and miR-21-5p was verified by RT-qPCR and luciferase assay. The functions of miR-21-5p in I/R-triggered injury were reassessed. PI3K/AKT/mTOR pathway was detected by Western blot. Down-regulated circYAP1 was observed in AKI blood samples and I/R-treated HK-2 cells. CircYAP1 overexpression expedited cell growth and weakened secretion of inflammatory factors and ROS generation in I/R-disposed cells. Besides, we found circYAP1 could sponge to miR-21-5p. Interestingly, miR-21-5p overexpression overturned the repressive effects of circYAP1 on cell injury. Moreover, PI3K/AKT/mTOR pathway was activated by circYAP1 via inhibiting miR-21-5p. We demonstrated that circYAP1 activated PI3K/AKT/mTOR pathway and secured HK-2 cells from I/R injury via sponging miR-21-5p.
环状 RNA YAP1（circYAP1）被报道参与胃癌的进展。然而，circYAP1 在急性肾损伤（AKI）中的作用仍不清楚。我们试图研究 circYAP1 对缺血/再灌注刺激的肾损伤的影响。通过在缺血/再灌注（I/R）环境中处理 HK-2 细胞来建立 AKI 模型。通过 RT-qPCR 评估 AKI 患者和 I/R 处理的 HK-2 细胞中 circYAP1 的表达。通过 CCK-8、流式细胞术、ELISA 和 ROS 测定来测试 circYAP1 对细胞活力、细胞凋亡、炎症细胞因子和 ROS 生成的影响。执行生物信息学分析以探索 miRNA 靶标。通过 RT-qPCR 和荧光素酶测定验证 circYAP1 与 miR-21-5p 的相关性。重新评估 miR-21-5p 在 I/R 触发损伤中的作用。通过 Western blot 检测 PI3K/AKT/mTOR 通路。在 AKI 血液样本和 I/R 处理的 HK-2 细胞中观察到下调的 circYAP1。circYAP1 的过表达加速了细胞生长，并减弱了 I/R 处理细胞中炎症因子的分泌和 ROS 的产生。此外，我们发现 circYAP1 可以与 miR-21-5p 结合。有趣的是，miR-21-5p 的过表达推翻了 circYAP1 对细胞损伤的抑制作用。此外，circYAP1 通过抑制 miR-21-5p 激活 PI3K/AKT/mTOR 通路。我们证明 circYAP1 通过与 miR-21-5p 结合激活 PI3K/AKT/mTOR 通路，保护 HK-2 细胞免受 I/R 损伤。
