Department of Respiratory and Asthma, Xi'an Children's Hospital , Xi'an, Shanxi, China.
Cell Cycle. 2020 Jun;19(11):1275-1284. doi: 10.1080/15384101.2020.1746874. Epub 2020 Apr 19.
YAP1 was previously reported to regulate the development of multiple tumors, angiogenesis included. Angiogenesis was a specific process of remodeling in asthma. In a recent study, YAP1 was correlated with the progression of asthma. However, the role of YAP1 in airway smooth muscle cell and the asthmatic airway angiogenesis was unclear. In the present study, we used cytokine-stimulated airway smooth muscle cells as asthma cell model in vitro. The results showed a significant up-regulation of YAP1 in asthmatic airway smooth muscle tissue and cytokine-stimulated asthmatic cell model by Western blot. The experimental results of YAP1 loss-of-function combined with STAT3 inhibitor (WP1066) showed that YAP1 knockdown inhibited the expression of VEGF by deactivating STAT3 in cytokine-stimulated ASM cells, which hindered the pro-angiogenesis ability of ASM cells. Besides, by combining prediction and binding site mutation along with luciferase reporter gene experiments, we confirmed direct binding between miR-375 and YAP1. Based on that, the decreased expression level of miR-375 was found to be correlated with the pathogenesis of asthma. Finally, miR-375 was verified to participate in the YAP1-regulated pro-angiogenesis ability of ASM cells. To sum up, we provided the evidence that YAP1 directly binds to miR-375 and takes part in the regulation of the pro-angiogenic ability of ASM cells by activating STAT3 and VEGF signaling.
YAP1 先前被报道可调节多种肿瘤的发展,包括血管生成。血管生成是哮喘中一种特定的重塑过程。最近的一项研究表明,YAP1 与哮喘的进展有关。然而,YAP1 在气道平滑肌细胞和哮喘气道血管生成中的作用尚不清楚。在本研究中,我们使用细胞因子刺激的气道平滑肌细胞作为体外哮喘细胞模型。Western blot 结果显示,哮喘气道平滑肌组织和细胞因子刺激的哮喘细胞模型中 YAP1 的表达显著上调。YAP1 功能丧失与 STAT3 抑制剂(WP1066)联合实验的结果表明,YAP1 敲低通过失活 STAT3 抑制细胞因子刺激的 ASM 细胞中 VEGF 的表达,从而抑制 ASM 细胞的促血管生成能力。此外,通过预测和结合位点突变以及荧光素酶报告基因实验,我们证实了 miR-375 与 YAP1 之间的直接结合。基于此,发现 miR-375 的表达水平降低与哮喘的发病机制有关。最后,验证了 miR-375 参与 YAP1 调节的 ASM 细胞促血管生成能力。总之,我们提供了证据表明,YAP1 直接结合 miR-375,并通过激活 STAT3 和 VEGF 信号通路参与 ASM 细胞的促血管生成能力的调节。