MiR-27a-3p/miR-27b-3p Promotes Neurofibromatosis Type 1 via Targeting of NF1


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.

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Neurofibromatosis type 1


Neurofibromin 1


Malignant peripheral nerve sheath tumors


Human Schwann cells


Un-translated region


Messenger RNA




Dulbecco’s modified Eagle’s medium


Fetal bovine serum


Real-time quantitative polymerase chain reaction


Cell Counting Kit-8








Terminal deoxynucleotidyl transferase dUTP nick-end labeling


RNA binding protein immunoprecipitation






Standard deviation


Analysis of variance


  1. An T et al (2019) Comparison of Alterations in miRNA Expression in Matched Tissue and Blood Samples during Spinal Cord Glioma Progression Scientific reports 9:9169. https://doi.org/10.1038/s41598-019-42364-x

    CAS  Article  PubMed  Google Scholar 

  2. Barkan B, Starinsky S, Friedman E, Stein R, Kloog Y (2006) The Ras inhibitor farnesylthiosalicylic acid as a potential therapy for neurofibromatosis type 1 Clinical cancer research : an official journal of the American Association for Cancer Research 12:5533–5542. https://doi.org/10.1158/1078-0432.Ccr-06-0792

  3. Cappione AJ, French BL, Skuse GR (1997) A potential role for NF1 mRNA editing in the pathogenesis of NF1 tumors. Am J Hum Genet 60:305–312

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Cimino PJ, Gutmann DH (2018) Neurofibromatosis type 1. Handb Clin Neurol 148:799–811. https://doi.org/10.1016/b978-0-444-64076-5.00051-x

    Article  PubMed  Google Scholar 

  5. Frassanito MA et al (2019) Bone marrow fibroblasts overexpress miR-27b and miR-214 in step with multiple myeloma progression, dependent on tumour cell-derived exosomes. J Pathol 247:241–253. https://doi.org/10.1002/path.5187

    CAS  Article  PubMed  Google Scholar 

  6. Gong M, Ma J, Li M, Zhou M, Hock JM, Yu X (2012) MicroRNA-204 critically regulates carcinogenesis in malignant peripheral nerve sheath tumors. Neuro-oncology 14:1007–1017. https://doi.org/10.1093/neuonc/nos124

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. Gutmann DH et al (1997) The diagnostic evaluation and multidisciplinary management of neurofibromatosis 1 and neurofibromatosis 2. Jama 278:51–57

  8. Hoffmeyer S, Assum G, Griesser J, Kaufmann D, Nurnberg P, Krone W (1995) On unequal allelic expression of the neurofibromin gene in neurofibromatosis type 1. Hum Mol Genet 4:1267–1272. https://doi.org/10.1093/hmg/4.8.1267

    CAS  Article  PubMed  Google Scholar 

  9. Li Y et al (1995) Genomic organization of the neurofibromatosis 1 gene (NF1). Genomics 25:9–18. https://doi.org/10.1016/0888-7543(95)80104-t

    CAS  Article  PubMed  Google Scholar 

  10. Masliah-Planchon J et al (2013) MicroRNAome profiling in benign and malignant neurofibromatosis type 1-associated nerve sheath tumors: evidences of PTEN pathway alterations in early NF1 tumorigenesis BMC. Genomics 14:473. https://doi.org/10.1186/1471-2164-14-473

    CAS  Article  PubMed  Google Scholar 

  11. Metheny LJ, Cappione AJ, Skuse GR (1995) Genetic and epigenetic mechanisms in the pathogenesis of neurofibromatosis type I. J Neuropathol Exp Neurol 54:753–760. https://doi.org/10.1097/00005072-199511000-00001

    CAS  Article  PubMed  Google Scholar 

  12. Ottaviani S et al (2018) TGF-β induces miR-100 and miR-125b but blocks let-7a through LIN28B controlling PDAC progression. Nat Commun 9:1845. https://doi.org/10.1038/s41467-018-03962-x

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Riccardi VM, Womack JE, Jacks T (1994) Neurofibromatosis and related tumors. Natural occurrence and animal models. Am J Pathol 145:994–1000

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Rouhi A, Mager DL, Humphries RK, Kuchenbauer F (2008) MiRNAs, epigenetics, and cancer Mammalian genome : official journal of the International Mammalian Genome Society 19:517–525. https://doi.org/10.1007/s00335-008-9133-x

  15. Skuse GR, Cappione AJ (1997) NA processing and clinical variability in neurofibromatosis type I (NF1). Hum Mol Genet 6:1707–1712

    CAS  Article  Google Scholar 

  16. Sun X et al (2017) Glioma stem cells-derived exosomes promote the angiogenic ability of endothelial cells through miR-21/VEGF signal. Oncotarget 8:36137–36148. https://doi.org/10.18632/oncotarget.16661

    Article  PubMed  PubMed Central  Google Scholar 

  17. Wang X et al (2019) LncRNA NEAT1 promotes extracellular matrix accumulation and epithelial-to-mesenchymal transition by targeting miR-27b-3p and ZEB1 in diabetic nephropathy. J Cell Physiol 234:12926–12933. https://doi.org/10.1002/jcp.27959

    CAS  Article  PubMed  Google Scholar 

  18. Xu Q, Cui Y, Luan J, Zhou X, Li H, Han J (2018) Exosomes from C2C12 myoblasts enhance osteogenic differentiation of MC3T3-E1 pre-osteoblasts by delivering miR-27a-3p . Biochem Biophys Res Commun 498:32–37. https://doi.org/10.1016/j.bbrc.2018.02.144

    CAS  Article  PubMed  Google Scholar 

  19. Xu W et al (2013) miR-24–3p and miR-27a-3p promote cell proliferation in glioma cells via cooperative regulation of MXI1. Int J Oncol 42:757–766. https://doi.org/10.3892/ijo.2012.1742

    CAS  Article  PubMed  Google Scholar 

  20. Yang X et al (2019) MiR-27b-3p promotes migration and invasion in colorectal cancer cells by targeting HOXA10. Bioscience Rep 39. https://doi.org/10.1042/bsr20191087

  21. Zhang P et al (2014) EZH2-miR-30d-KPNB1 pathway regulates malignant peripheral nerve sheath tumour cell survival and tumourigenesis. J Pathol 232:308–318. https://doi.org/10.1002/path.4294

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. Zhao B, Zhang J, Chen X, Xu H, Huang B (2019) Mir-26b inhibits growth and resistance to paclitaxel chemotherapy by silencing the CDC6 gene in gastric cancer Archives of medical science : AMS 15:498–503. https://doi.org/10.5114/aoms.2018.73315

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Correspondence to Quantang Pang.

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The human tissues utilized in the study were collected under the subject protocol approved by Institutional Review Boards of Rongcheng People’s Hospital of Shandong Province.

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Supplementary file1 (TIF 1316 KB) Figure S1. A. The expression of miR-27a-3p/miR-27b-3p in normal HSC was analyzed after transfection with miR-27a-3p/miR-27b-3p mimics. B. The cell vitality was studied by CCK8 after upregulation of miR-27b-3p and miR-27a-3p. C. Transwell assay was carried out to assess the migratory ability of the transfected normal HSC. **P  <  0.01

Supplementary file2 (TIF 2089 KB) Figure S2. A. The expression of miR-27a-3p/miR-27b-3p in ST88-14 was analyzed after transfection with miR-27a-3p/miR-27b-3p mimics. B–C. The cell vitality and proliferation were studied by CCK8 and EdU assays, respectively, after upregulation of miR-27b-3p and miR-27a-3p. D. Transwell assay was carried out to assess the migratory ability of the transfected ST88-14. **P  <  0.01

Supplementary file3 (TIF 1677 KB) Figure S3. A. The interference effectiveness of miR-27a-3p/miR-27b-3p inhibitor in ST88-14 was detected by RT-qPCR analysis. B–F. A series of functional experiments were performed to elucidate the effects of miR-27a-3p/miR-27b-3p inhibitor on proliferation, apoptosis, and migration by CCK8 (B), TUNEL (C), flow cytometry analysis (D), caspase-3 detection assay (E), and transwell assay (F). **P <  0.01.

Supplementary file4 (TIF 2903 KB) Figure S4. A. RT-qPCR assay was performed to study the expression of NF1 mRNA in the transfected ST88-14 with miR-27a-3p/miR-27b-3p mimics. B. RNA pull-down assay was completed to elucidate the molecular relationship between miR-27a-3p/miR-27b-3p and NF1. C. RIP assay was utilized to verify the interaction between target miRNAs and NF1. D-I. The rescue impact of NF1 depletion on miR-27b-3p/miR-27a-3p inhibitor-induced cell viability, proliferation, apoptosis, and migration phenotype was studied via CCK8 (D), EdU assay (E), TUNEL (F), flow cytometry analysis (G), caspase-3 detection assay (H), and transwell assay (I). **P  <  0.01.

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Lu, H., Liu, P. & Pang, Q. MiR-27a-3p/miR-27b-3p Promotes Neurofibromatosis Type 1 via Targeting of NF1 . J Mol Neurosci (2021). https://doi.org/10.1007/s12031-020-01779-2

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  • Neurofibromatosis type 1
  • NF1
  • miR-27a-3p
  • miR-27b-3p