Experimental verification of a conserved intronic microRNA located in the human TrkC gene with a cell type-dependent apoptotic function

Cell Mol Life Sci. 2015 Jul;72(13):2613-25. doi: 10.1007/s00018-015-1868-4. Epub 2015 Mar 14.

Abstract

Tropomyosin receptor kinase C (TrkC) is involved in cell survival, apoptosis induction and tumorigenesis. We hypothesized that, similar to p75(NTR) receptor, some of the diverse functions of TrkC could be mediated by a microRNA (miRNA) embedded within the gene. Here, we experimentally verified the expression and processing of two bioinformatically predicted miRNAs named TrkC-miR1-5p and TrkC-miR1-3p. Transfecting a DNA fragment corresponding to the TrkC-premir1 sequence in HEK293t cells caused ~300-fold elevation in the level of mature TrkC-miR1 and also a significant downregulation of its predicted target genes. Furthermore, endogenous TrkC-miR1 was detected in several cell lines and brain tumors confirming its endogenous generation. Furthermore, its orthologous miRNA was detected in developing rat brain. Accordingly, TrkC-miR1 expression was increased during the course of neural differentiation of NT2 cell, whereas its suppression attenuated NT2 differentiation. Consistent with opposite functions of TrkC, TrkC-miR1 overexpression promoted survival and apoptosis in U87 and HEK293t cell lines, respectively. In conclusion, our data report the discovery of a new miRNA with overlapping function to TrkC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Apoptosis / genetics*
  • Azacitidine
  • Brain / growth & development
  • Brain / metabolism*
  • Cell Differentiation / physiology
  • Cell Line, Tumor
  • Computational Biology
  • DNA Primers / genetics
  • Gene Expression Regulation / genetics*
  • HEK293 Cells
  • Humans
  • Introns / genetics*
  • MicroRNAs / genetics*
  • Neurons / metabolism
  • Neurons / physiology
  • Rats
  • Receptor, trkC / genetics*
  • Receptor, trkC / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Primers
  • MicroRNAs
  • Receptor, trkC
  • Azacitidine