Journal of Biosciences

, Volume 42, Issue 1, pp 23–30 | Cite as

Overexpression of hsa-miR-939 follows by NGFR down-regulation and apoptosis reduction

  • Fahimeh Hosseini Aghdaei
  • Bahram M Soltani
  • Sadat Dokanehiifard
  • Seyed Javad Mowla
  • Masoud Soleimani


Neurotrophin receptors play a crucial role in neuronal survival, differentiation and regeneration. Nerve growth factor receptor (NGFR) or P75 NTR is a neurotrophin receptor that is involved in many pathological conditions including cancers. Genetic factors that are involved in regulation of neurotrophin receptors are under intense investigation. MiRNAs are novel regulators of signalling pathways that are candidates for regulation of neurotrophin receptors. Computational programs predicted that NGFR gene is a bona fide target for hsa-miR-939. RT-qPCR, Western analysis and dual luciferase assay evidences indicated that NGFR transcript is targeted by hsa-miR-939. Also, hsa-miR-939 overexpression brought about down-regulation of NGFR expression in U87 cell line, followed by cell death rate reduction, detected by flow cytometry. Taken together, here for the first time, hsa-miR-939 is introduced as a novel key regulator of NGFR expression and its involvement in cell death/survival processes is suggested.


Apoptosis hsa-miR-939 luciferase assay neurotrophin signalling NGFR 

Abbreviations used


MiRNA recognition element


nerve growth factor receptor


precursor miRNAs


untranslated region



The authors thank Dr Saman Hosseinkhani for his kind advice. This work was supported by TMU, ISTI and INSF financial aids.


  1. Allen M, Bjerke M, Edlund H, Nelander S and Westermark B 2016 Origin of the U87MG glioma cell line: good news and bad news. Sci. Transl. Med. 8 354re3–re3CrossRefPubMedGoogle Scholar
  2. Ambros V 2004 The functions of animal microRNAs. Nature 431 350–355CrossRefPubMedGoogle Scholar
  3. Blöchl A and Blöchl R 2007 A cell-biological model of p75NTR signaling. J. Neurochem. 102 289–305CrossRefPubMedGoogle Scholar
  4. Cantarella G, Lempereur L, Presta M, Ribatti D, Lombardo G, Lazarovici P, et al. 2002 Nerve growth factor-endothelial cell interaction leads to angiogenesis in vitro and in vivo. FASEB J. 16 1307–1309Google Scholar
  5. Chang C-C, Fang W-H, Chang H-A and Huang S-Y 2015 Functional Ser205Leu polymorphism of the nerve growth factor receptor (NGFR) gene is associated with vagal autonomic dysregulation in humans. Sci Rep 5 13136Google Scholar
  6. Cheng H-C, Sun Y, Lai L-C, Chen S-Y, Lee W-C, Chen J-H, et al. 2012 Genetic polymorphisms of nerve growth factor receptor (NGFR) and the risk of Alzheimer's disease. J. Negat. Results Biomed. 11 5CrossRefPubMedPubMedCentralGoogle Scholar
  7. Clark MJ, Homer N, O'Connor BD, Chen Z, Eskin A, Lee H, et al. 2010 U87MG decoded: the genomic sequence of a cytogenetically aberrant human cancer cell line. PLoS Genet. 6 e1000832CrossRefPubMedPubMedCentralGoogle Scholar
  8. Dennis G, Sherman B, Hosack D, Yang J, Gao W, Lane HC, et al. 2003 DAVID: database for annotation, visualization, and integrated discovery. Genome Biol. 4 P3CrossRefPubMedGoogle Scholar
  9. Du T and Zamore PD 2005 microPrimer: the biogenesis and function of microRNA. Development 132 4645–52CrossRefPubMedGoogle Scholar
  10. Dweep H, Sticht C, Pandey P and Gretz N 2011 miRWalk – database: prediction of possible miRNA binding sites by “walking” the genes of three genomes. J. Biomed. Inform. 44 839–47CrossRefPubMedGoogle Scholar
  11. Frade JM and Barde YA 1998 Nerve growth factor: two receptors, multiple functions. Bioessays 20 137–45CrossRefPubMedGoogle Scholar
  12. Friedman RC, Farh KK-H, Burge CB and Bartel DP 2009 Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19 92–105CrossRefPubMedPubMedCentralGoogle Scholar
  13. Galardi S, Mercatelli N, Giorda E, Massalini S, Frajese GV, Ciafrè SA, et al. 2007 miR-221 and miR-222 expression affects the proliferation potential of human prostate carcinoma cell lines by targeting p27Kip1. J. Biol. Chem. 282 23716–24CrossRefPubMedGoogle Scholar
  14. Gao X, Daugherty RL and Tourtellotte WG 2007 Regulation of low affinity neurotrophin receptor (p75NTR) by early growth response (Egr) transcriptional regulators. Mol. Cell. Neurosci. 36 501–14CrossRefPubMedPubMedCentralGoogle Scholar
  15. Gentry JJ, Barker PA, Carter BD 2004 The p75 neurotrophin receptor: multiple interactors and numerous functions; in Progress in brain research (eds) A Luigi, C Laura (Elsevier) pp 25–39Google Scholar
  16. Gramantieri L, Ferracin M, Fornari F, Veronese A, Sabbioni S, Liu C-G, et al. 2007 Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma. Cancer Res. 67 6092–9CrossRefPubMedGoogle Scholar
  17. Hefti F and Mash DC 1989 Localization of nerve growth factor receptors in the normal human brain and in Alzheimer's disease. Neurobiol. Aging 10 75–87CrossRefPubMedGoogle Scholar
  18. Hu Z, Luo J, Zhong S, Xue L, Chen Y and Fan R 2012 MicroRNAs expression in normal and dissected aortic tissue. Zhonghua Xin Xue Guan Bing Za Zhi. 40 406PubMedGoogle Scholar
  19. Irmady K, Jackman KA, Padow VA, Shahani N, Martin LA, Cerchietti L, et al. 2014 MiR-592 regulates the induction and cell death-promoting activity of p75NTR in neuronal ischemic injury. J. Neurosci. 34 3419–28CrossRefPubMedPubMedCentralGoogle Scholar
  20. Johnston ALM, Lun X, Rahn JJ, Liacini A, Wang L, Hamilton MG, et al. 2007 The p75 neurotrophin receptor is a central regulator of glioma invasion. PLoS Biol. 5 e212CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kent WJ 2002 BLAT—the BLAST-like alignment tool. Genome Res. 12 656–64CrossRefPubMedPubMedCentralGoogle Scholar
  22. Khvorova A, Reynolds A and Jayasena SD 2003 Functional siRNAs and miRNAs exhibit strand bias. Cell 115 209–16CrossRefPubMedGoogle Scholar
  23. Khwaja F, Tabassum A, Allen J and Djakiew D 2006 The p75 NTR tumor suppressor induces cell cycle arrest facilitating caspase mediated apoptosis in prostate tumor cells. Biochem. Biophys. Res. Commun. 341 1184–92CrossRefPubMedGoogle Scholar
  24. Krygier S and Djakiew D 2001 Molecular characterization of the loss of p75NTR expression in human prostate tumor cells. Mol. Carcinog. 31 46–55CrossRefPubMedGoogle Scholar
  25. Kumar A, Sinha RA, Tiwari M, Pal L, Shrivastava A, Singh R, et al. 2006 Increased pro-nerve growth factor and p75 neurotrophin receptor levels in developing hypothyroid rat cerebral cortex are associated with enhanced apoptosis. Endocrinology 147 4893–903CrossRefPubMedGoogle Scholar
  26. Lewis BP, Burge CB and Bartel DP 2005 Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are MicroRNA targets. Cell 120 15–20CrossRefPubMedGoogle Scholar
  27. Maragkakis M, Reczko M, Simossis VA, Alexiou P, Papadopoulos GL, Dalamagas T, et al. 2009 DIANA-microT web server: elucidating microRNA functions through target prediction. Nucleic Acids Res. 37 W273–W6CrossRefPubMedPubMedCentralGoogle Scholar
  28. Mestdagh P, Van Vlierberghe P, De Weer A, Muth D, Westermann F, Speleman F, et al. 2009 A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol. 10 R64CrossRefPubMedPubMedCentralGoogle Scholar
  29. Molloy NH, Read DE and Gorman AM 2011 Nerve growth factor in cancer cell death and survival. Cancer 3 510–30CrossRefGoogle Scholar
  30. Mufson EJ and Kordower JH 1992 Cortical neurons express nerve growth factor receptors in advanced age and Alzheimer disease. Proc. Natl. Acad. Sci. 89 569–73CrossRefPubMedPubMedCentralGoogle Scholar
  31. Nakabayashi H, Taketa K, Miyano K, Yamane T and Sato J 1982 Growth of human hepatoma cell lines with differentiated functions in chemically defined medium. Cancer Res. 42 3858–63PubMedGoogle Scholar
  32. Ramos A, Chi Ho W, Forte S, Dickson K, Boutilier J, Favell K, et al. 2007 Hypo-Osmolar stress induces p75NTR expression by activating Sp1-dependent transcription. J. Neurosci. 27 1498–506CrossRefPubMedGoogle Scholar
  33. Rani S, Gately K, Crown J, O’Byrne K and O’Driscoll L 2013 Global analysis of serum microRNAs as potential biomarkers for lung adenocarcinoma. Cancer Biol. Ther. 14 1CrossRefGoogle Scholar
  34. Reichardt LF 2006 Neurotrophin-regulated signalling pathways. Philos. Trans. R. Soc., B. 361 1545–64CrossRefGoogle Scholar
  35. Rocha AS, Risberg B, Magalhães J, Trovisco V, de Castro IV, Lazarovici P, et al. 2006 The p75 neurotrophin receptor is widely expressed in conventional papillary thyroid carcinoma. Hum. Pathol. 37 562–8CrossRefPubMedGoogle Scholar
  36. Rodriguez-Tebar A, Dechant G and Barde Y-A 1990 Binding of brain-derived neurotrophic factor to the nerve growth factor receptor. Neuron 4 487–92CrossRefPubMedGoogle Scholar
  37. Roux PP and Barker PA 2002 Neurotrophin signaling through the p75 neurotrophin receptor. Prog. Neurobiol. 67 203–33CrossRefPubMedGoogle Scholar
  38. Salis MB, Graiani G, Desortes E, Caldwell RB, Madeddu P and Emanueli C 2004 Nerve growth factor supplementation reverses the impairment, induced by Type 1 diabetes, of hindlimb post-ischaemic recovery in mice. Diabetologia 47 1055–63CrossRefPubMedGoogle Scholar
  39. Scarpini E, Conti G, Chianese L, Baron P, Pizzul S, Basellini A, et al. 1996 Induction of p75NGFR in human diabetic neuropathy. J. Neurol. Sci. 135 55–62CrossRefPubMedGoogle Scholar
  40. Troy CM, Friedman JE and Friedman WJ 2002 Mechanisms of p75-mediated death of hippocampal neurons role of caspases. J. Biol. Chem. 277 34295–302CrossRefPubMedGoogle Scholar
  41. Watanabe Y, Tomita M, Kanai A 2007 Computational methods for MicroRNA target prediction; in: Methods in enzymology (eds) JR John, JH Gregory (Academic Press) pp 65–86Google Scholar
  42. Zhang X, Chen C, Wu M, Chen L, Zhang J, Zhang X, et al. 2012 Plasma microRNA profile as a predictor of early virological response to interferon treatment in chronic hepatitis B patients. Antivir. Ther. 17 1243–53CrossRefPubMedGoogle Scholar

Copyright information

© Indian Academy of Sciences 2017

Authors and Affiliations

  • Fahimeh Hosseini Aghdaei
    • 1
  • Bahram M Soltani
    • 1
  • Sadat Dokanehiifard
    • 1
  • Seyed Javad Mowla
    • 1
  • Masoud Soleimani
    • 2
  1. 1.Molecular Genetics Department, Faculty of Biological SciencesTarbiat Modares UniversityTehranIran
  2. 2.Hematology Department, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran

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