Long Noncoding RNA Rps4l Mediates the Proliferation of Hypoxic Pulmonary Artery Smooth Muscle Cells
Long Noncoding RNA Rps4l Mediates the Proliferation of Hypoxic Pulmonary Artery Smooth Muscle Cells
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DOI：
10.1161/hypertensionaha.120.14644
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发表时间：
2020-10-01
期刊：
影响因子：
8.3
通讯作者：
Zhu, Daling
中科院分区：
文献类型：
--
作者：
Liu, Ying;Zhang, Hongyue;Zhu, Daling
肺动脉高压（PH）是一种罕见的致命性疾病，涉及由肺动脉平滑肌细胞（PASMCs）增殖增强介导的肺动脉血管重塑。长链非编码RNA是具有不同细胞功能的调节分子的一个亚类，但它们在PH中的作用在很大程度上尚未探索。我们的目的是鉴定和确定参与低氧诱导的PH和PASMC增殖的长非编码RNA的功能。缺氧小鼠模型中的RNA测序鉴定了缺氧调节的长非编码RNA，包括Rps 4l。Rps 41在PH模型小鼠和缺氧PASMCs中的表达显著降低。通过RNA荧光原位杂交和核质RNA定量检测Rps 41的亚细胞定位。rps 4l过表达挽救了肺动脉高压特征，如右心室肥大、右心室收缩压、血流动力学、心功能和血管重构所证明的。在细胞水平，Rps 41过表达削弱细胞活力和增殖，并抑制细胞周期进程。潜在的Rps 4l结合蛋白通过RNA下拉，然后通过质谱，RNA免疫沉淀，和微量热泳鉴定。这些结果表明Rps 41与ILF 3（白细胞介素增强子结合因子3）的稳定性相关并影响ILF 3的稳定性。Rps 41进一步调节HIF-1 α的水平，从而导致缺氧诱导的PASMC增殖和迁移。我们的研究结果表明，在缺氧PASMCs，Rps 41的表达下降，由于缺氧的调节。这种减少通过ILF 3/HIF-1 α影响PASMCs的增殖、迁移和细胞周期进程。这些结果为进一步研究缺氧性PH的病理机制提供了理论基础，并可能为开发新的治疗方法提供见解。
Pulmonary hypertension (PH) is a rare and fatal disorder involving the vascular remodeling of pulmonary arteries mediated by the enhanced proliferation of pulmonary artery smooth muscle cells (PASMCs). Long noncoding RNAs are a subclass of regulatory molecules with diverse cellular functions, but their role in PH remains largely unexplored. We aimed to identify and determine the functions of long noncoding RNAs involved in hypoxia-induced PH and PASMC proliferation. RNA sequencing in a hypoxic mouse model identified hypoxia-regulated long noncoding RNAs, including Rps4l. Rps4l expression was significantly reduced in PH-model mice and hypoxic PASMCs. The subcellular localization of Rps4l was detected by RNA fluorescence in situ hybridization and quantification of nuclear/cytoplasmic RNA. Rps4l overexpression rescued pulmonary arterial hypertension features, as demonstrated by right ventricle hypertrophy, right ventricular systolic pressure, hemodynamics, cardiac function, and vascular remodeling. At the cellular level, Rps4l overexpression weakened cell viability and proliferation and suppressed cell cycle progression. Potential Rps4l-binding proteins were identified via RNA pull-down followed by mass spectrometry, RNA immunoprecipitation, and microscale thermophoresis. These results indicated that Rps4l is associated with and affects the stabilization of ILF3 (interleukin enhancer-binding factor 3). Rps41 further regulates the levels of HIF-1 alpha and consequently leads to hypoxia-induced PASMC proliferation and migration. Our results showed that in hypoxic PASMCs, Rps4l expression decreases due to regulation by hypoxia. This decrease affects the proliferation, migration, and cell cycle progression of PASMCs through ILF3/HIF-1 alpha. These results provide a theoretical basis for further investigations into the pathological mechanism of hypoxic PH and may provide insight for the development of novel treatments.