Over-expression of miR-125a-5p inhibits proliferation in C2C12 myoblasts by targeting E2F3

Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):244-9. doi: 10.1093/abbs/gmv006. Epub 2015 Mar 1.

Abstract

MicroRNAs (miRNAs) are a class of small non-coding RNAs of 20-25 nucleotides in length. It has been shown that miRNAs play important roles in the proliferation of many types of cells, including myoblasts. In this study, we used real-time quantitative polymerase chain reaction, western blotting, EdU, flow cytometry, and CCK-8 assay to explore the role of miR-125a-5p during the proliferation of C2C12 myoblasts. It was found that the expression of miR-125a-5p was decreased during C2C12 myoblast proliferation. Over-expression of miR-125a-5p inhibited C2C12 myoblast proliferation as indicated by EdU staining, flow cytometry, and CCK8 assay. It was also found that miR-125a-5p could negatively regulate E2F3 expression at posttranscriptional level, via a specific target site in the 3' untranslated region. Knockdown of E2F3 showed a similar inhibitory effect on C2C12 myoblast proliferation. Thus, our findings suggest that miR-125a-5p may act as a negative regulator of C2C12 myoblast proliferation by targeting E2F3.

Keywords: E2F3; miR-125a-5p, proliferation; microRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • Animals
  • Blotting, Western
  • Cell Line
  • Cell Proliferation / genetics*
  • Cyclin D1 / genetics
  • Cyclin D1 / metabolism
  • DNA Replication / genetics
  • E2F3 Transcription Factor / genetics*
  • E2F3 Transcription Factor / metabolism
  • G1 Phase Cell Cycle Checkpoints / genetics
  • Gene Expression*
  • Gene Knockdown Techniques
  • Mice
  • MicroRNAs / genetics*
  • Microscopy, Fluorescence
  • Mutation
  • Myoblasts / cytology
  • Myoblasts / metabolism*
  • RNA Interference
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • 3' Untranslated Regions
  • E2F3 Transcription Factor
  • E2f3 protein, mouse
  • MIRN1202 microRNA, human
  • MicroRNAs
  • Mirn125 microRNA, mouse
  • Cyclin D1