Department of Neurology, Rongcheng People's Hospital of Shandong Province, Rongcheng, 264300, Shandong, China.
Department of Orthopedic, Rongcheng People's Hospital of Shandong Province, Rongcheng, 264300, Shandong, China.
J Mol Neurosci. 2021 Nov;71(11):2353-2363. doi: 10.1007/s12031-020-01779-2. Epub 2021 Feb 11.
The dysregulation of microRNAs (miRNAs) is a crucial molecular signature of disease development. The potential implication of miRNAs in neurofibromatosis type 1 (NF1) remains poorly investigated. The expression levels of miR-27a-3p, miR-27b-3p, and neurofibromin 1 (NF1) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) analysis. The functional roles of miR-27a-3p and miR-27b-3p in NF1 were explored by CCK8 (Cell Counting Kit-8), 5-ethynyl-2'-deoxyuridine (EdU), terminal deoxynucleoitidyl transferase dUTP nick-end labeling (TUNEL), and transwell assays. Luciferase reporter, RNA pull-down, and RNA binding protein immunoprecipitation (RIP) assays were employed to study the probable target relationship between miRNA and messenger RNA (mRNA). MiR-27b-3p and miR-27a-3p were upregulated in dermal and plexiform human Schwann cells (HSC) from NF1 neurofibromas as well as cell lines of malignant peripheral nerve sheath tumors (MPNSTs). MiR-27a-3p/miR-27b-3p mimics promoted the proliferative, migratory, and invasive ability of dermal HSC and MPNST cell ST88-14, while inhibiting the apoptotic capacity. MiR-27a-3p/miR-27b-3p inhibitors elicited the opposite impacts on the above cellular behaviors in dermal HSC and ST88-14. Intriguingly, NF1 was revealed to be the target of both miR-27a-3p and miR-27b-3p, and was negatively modulated by them. MiR-27a-3p/miR-27b-3p upregulation suppressed the expression of NF1 in dermal HSC and ST88-14. Furthermore, NF1 depletion counterbalanced the functional alteration induced by miR-27a-3p/miR-27b-3p inhibition. Our study suggests that both miR-27b-3p and miR-27a-3p are involved in upstream molecular activity responsible for the depletion of NF1, representing promising targets for therapeutic application in NF1.
miRNAs(微小 RNA)的失调是疾病发展的一个关键分子特征。miRNAs 在神经纤维瘤病 1 型(NF1)中的潜在作用仍未得到充分研究。通过实时定量聚合酶链反应(RT-qPCR)分析检测 miR-27a-3p、miR-27b-3p 和神经纤维瘤蛋白 1(NF1)的表达水平。通过 CCK8(细胞计数试剂盒-8)、5-乙炔基-2'-脱氧尿苷(EdU)、末端脱氧核苷酸转移酶 dUTP 缺口末端标记(TUNEL)和 Transwell 测定法探索 miR-27a-3p 和 miR-27b-3p 在 NF1 中的功能作用。荧光素酶报告、RNA 下拉和 RNA 结合蛋白免疫沉淀(RIP)测定法用于研究 miRNA 和信使 RNA(mRNA)之间可能的靶标关系。NF1 神经纤维瘤中的皮肤和丛状人雪旺细胞(HSC)以及恶性外周神经鞘肿瘤(MPNST)细胞系中,miR-27b-3p 和 miR-27a-3p 上调。miR-27a-3p/miR-27b-3p 模拟物促进了皮肤 HSC 和 MPNST 细胞 ST88-14 的增殖、迁移和侵袭能力,同时抑制了凋亡能力。miR-27a-3p/miR-27b-3p 抑制剂在皮肤 HSC 和 ST88-14 中引起了上述细胞行为的相反影响。有趣的是,NF1 被揭示为 miR-27a-3p 和 miR-27b-3p 的靶标,并受其负调控。miR-27a-3p/miR-27b-3p 上调抑制了皮肤 HSC 和 ST88-14 中 NF1 的表达。此外,NF1 耗竭平衡了 miR-27a-3p/miR-27b-3p 抑制诱导的功能改变。我们的研究表明,miR-27b-3p 和 miR-27a-3p 都参与了负责 NF1 耗竭的上游分子活性,代表了 NF1 治疗应用的有前途的靶点。