let-7b-5p 通过下调 IGF1 抑制肺动脉平滑肌细胞的增殖和迁移。
let-7b-5p suppresses the proliferation and migration of pulmonary artery smooth muscle cells via down-regulating IGF1.
机构信息
Department of Respiratory Medicine, The Second People's Hospital of Hefei, Hefei Hospital Affiliated to Anhui Medical University, Hefei, Anhui 230011, China.
Department of Respiratory Medicine, The Second People's Hospital of Hefei, Hefei Hospital Affiliated to Anhui Medical University, Hefei, Anhui 230011, China.
出版信息
Clinics (Sao Paulo). 2022 May 27;77:100051. doi: 10.1016/j.clinsp.2022.100051. eCollection 2022.
OBJECTIVES
Some previous studies indicated that the excessive proliferation and migration of Pulmonary Artery Smooth Muscle Cells (PASMCs) could be observed in pulmonary artery intima after Pulmonary Embolism (PE) occurred. In addition, recent studies identified some miRNAs that are differentially expressed in the blood of PE patients, which might be used as a diagnostic biomarker for PE, including let-7a-5p, let-7b-5p, and miR-150-5p. Hence, the authors sought to explore the effects of let-7b-5p in PASMC proliferation and migration and the corresponding regulatory mechanism.
METHODS
Platelet-Derived Growth Factor (PDGF) was utilized to induce the hyper-proliferation model in PASMCs. The mRNA and protein expression levels were detected by RT-qPCR and western blot, respectively. The proliferation of PASMCs was evaluated by the detection of PCNA expression, as well as CCK-8 and Edu assays. Wound healing and Transwell assays were exploited to assess the migration ability of PASMCs. The targets of let-7b-5p were predicted based on two bioinformatics online tools. Dual-luciferase and Ago2 pull-down assays were applied to confirm the interaction between let-7b-5p and IGF1.
RESULTS
40 ng/mL PDGF was selected as the optimal concentration to induce PASMCs. let-7b-5p mimics suppressed the proliferation and migration of PDGF-induced PASMCs, while let-7b-5p inhibitor led to the opposite result. In further mechanism exploration, IGF1 was predicted and confirmed as the direct target gene of let-7b-5p. The promotion role of IGF1 overexpression on the proliferation and migration of PDGF-induced PASMCs was dramatically countered by let-7b-5p mimics.
CONCLUSION
let-7b-5p prohibits the proliferation and migration of PDGF-induced PASMCs by modulating IGF1.
目的
一些先前的研究表明，在肺栓塞（PE）发生后，肺动脉内膜中的肺动脉平滑肌细胞（PASMC）可观察到过度增殖和迁移。此外，最近的研究确定了一些在 PE 患者血液中表达差异的 miRNAs，它们可能被用作 PE 的诊断生物标志物，包括 let-7a-5p、let-7b-5p 和 miR-150-5p。因此，作者试图探讨 let-7b-5p 对 PASMC 增殖和迁移的影响及其相应的调节机制。
方法
血小板衍生生长因子（PDGF）用于诱导 PASMC 过度增殖模型。分别通过 RT-qPCR 和 Western blot 检测 mRNA 和蛋白表达水平。通过检测 PCNA 表达以及 CCK-8 和 Edu 测定来评估 PASMC 的增殖。利用划痕愈合和 Transwell 测定评估 PASMC 的迁移能力。利用两个生物信息学在线工具预测 let-7b-5p 的靶标。双荧光素酶和 Ago2 下拉测定用于证实 let-7b-5p 与 IGF1 之间的相互作用。
结果
选择 40ng/mL PDGF 作为诱导 PASMC 的最佳浓度。let-7b-5p 模拟物抑制 PDGF 诱导的 PASMC 增殖和迁移，而 let-7b-5p 抑制剂则导致相反的结果。在进一步的机制探索中，预测并证实 IGF1 是 let-7b-5p 的直接靶基因。IGF1 过表达对 PDGF 诱导的 PASMC 增殖和迁移的促进作用被 let-7b-5p 模拟物显著抵消。
结论
let-7b-5p 通过调节 IGF1 抑制 PDGF 诱导的 PASMC 增殖和迁移。
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