Inhibitory effect of NBL1 on PDGF-BB-induced human PASMC proliferation through blockade of PDGFβ-p38MAPK pathway.
Inhibitory effect of NBL1 on PDGF-BB-induced human PASMC proliferation through blockade of PDGFβ-p38MAPK pathway.
复制标题
NBL1 通过阻断 PDGFβ-p38MAPK 途径对 PDGF-BB 诱导的人 PASMC 增殖的抑制作用
DOI：
10.1042/bsr20160199
复制
发表时间：
2016-08
影响因子：
4
通讯作者：
Wei Y
中科院分区：
文献类型：
--
作者：
Cui C;Zhang H;Guo LN;Zhang X;Meng L;Pan X;Wei Y
肺动脉重构是肺动脉高压（PAH）病理过程中的一个重要特征。此外，肺动脉平滑肌细胞（PASMCs）的过度增殖在肺动脉重塑的发病机制中起着关键作用。神经母细胞瘤致瘤性抑制因子1（NBL 1）先前已显示可诱导肿瘤细胞的生长抑制。然而，NBL 1在调节人PASMC增殖中的作用仍不清楚。在培养的人PASMCs中，我们观察到NBL 1对血小板衍生生长因子（PDGF）-BB诱导的细胞生长、DNA合成和增殖细胞核抗原（PCNA）表达的剂量依赖性抑制作用，分别通过MTS分析、5-乙炔基-2-脱氧尿苷（EdU）分析和western blot测定。采用免疫印迹和免疫共沉淀法（IP）检测细胞周期正调控因子cyclin D1、cyclin E、CDK 2、CDK 4和CDK 6的表达和活性，以及细胞周期负调控因子p21和p27的表达和活性。我们的研究结果表明，NBL 1诱导的生长抑制与PDGF-BB处理的人PASMCs中细胞周期蛋白D1-CDK 4活性的降低和p27磷酸化的降低有关。通过使用磷特异性抗体的蛋白质印迹，我们进一步证明NBL 1诱导的生长抑制是通过阻断上游PDGF受体β（PDGFRβ）-p38丝裂原活化蛋白激酶（MAPK）介导的。结论：NBL 1可抑制PDGF-BB诱导的人PASMC增殖，其机制可能是通过阻断PDGFRβ-p38 MAPK信号通路，降低cyclin D1-CDK 4活性，上调p27的磷酸化水平。我们的研究结果可能为PAH提供一个潜在的治疗靶点。
Pulmonary artery remodelling is a key feature in the pathological progress of pulmonary arterial hypertension (PAH). Moreover, excessive proliferation of pulmonary arterial smooth muscle cells (PASMCs) plays a critical role in the pathogenesis of pulmonary artery remodelling. Neuroblastoma suppressor of tumorigenicity 1 (NBL1) has been previously shown to induce growth inhibition in tumour cells. However, the effect of NBL1 in the regulation of human PASMC proliferation remains unclear. In cultured human PASMCs, we observed a dose-dependent inhibitory effect of NBL1 on platelet derived growth factor (PDGF)-BB-induced cell growth, DNA synthesis and proliferating cell nuclear antigen (PCNA) expression, as measured by MTS assay, 5-ethynil-2-deoxyuridine (EdU) analysis and western blots respectively. We also detected the expression and activities of cell-cycle positive regulators (cyclin D1, cyclin E, CDK2, CDK4 and CDK6) and negative regulators (p21 and p27) in human PASMCs by western blots and co-immuoprecipitation (IP). Our results show that NBL1-induced growth suppression is associated with the decreased activity of cyclin D1–CDK4 and the decreased phosphorylation of p27 in PDGF-BB-treated human PASMCs. By western blots using the phosphor-specific antibodies, we further demonstrated that NBL1 induced growth suppression is mediated by blockade of the up-stream PDGF-receptor β (PDGFRβ)-p38 mitogen-activated protein kinase (MAPK). In conclusion, our results suggest that NBL1 could inhibit PDGF-BB-induced human PASMC proliferation, and the underlying mechanism is associated with the decreased cyclin D1–CDK4 activity and up-regulated p27 by decreasing the phosphorylation of p27 via blockade of PDGFRβ-p38MAPK signal cascade. Our findings may provide a potential therapeutic target for PAH.