miR-199a-5p regulates the expression of metastasis-associated genes in B16F10 melanoma cells

Int J Clin Exp Pathol. 2014 Sep 15;7(10):7182-90. eCollection 2014.

Abstract

MicroRNAs are regulatory factors that play important roles in tumor development, invasion and metastasis. Previously, we showed that miR-199a is abnormally expressed in clinical melanoma specimens and expression was closely associated with clinical features of metastasis. However, the exact molecular mechanisms by which miR-199a-5p influences melanoma invasion and metastasis remains unclear. In this study, we investigated gene expression changes of metastasis-associated genes in B16F10 melanoma cells following targeted silencing or overexpression of miR-199a-5p, using mouse tumor metastasis PCR arrays. Comparison of gene expression changes in miR-199a-5p-silenced versus overexpressing cells identified a set of upregulated genes (> 2-fold) including Cd44, Cdh1, Cxcr4, Etv4, Fxyd5, Rpsa, Mmp3, Myc, Rb1, Tcf20, Hprt1, Actb1 and downregulated genes (> 2-fold) including Ctsk, Itga7 and Tnfsf10. Regulation of a subset of these genes (Myc, Tnfsf10 and Cd44) following miR-199a-5p silencing or overexpression was validated by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. In conclusion, our study demonstrates that miR-199a-5p regulates melanoma metastasis-related genes, and may provide a basis for the development of novel, molecularly targeted drugs.

Keywords: Melanoma; metastasis PCR array; miR-199a-5p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Neoplastic*
  • Gene Silencing
  • Melanoma, Experimental / genetics*
  • Melanoma, Experimental / metabolism
  • Melanoma, Experimental / secondary*
  • Mice
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Neoplasm Invasiveness
  • Oligonucleotide Array Sequence Analysis
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Skin Neoplasms / genetics*
  • Skin Neoplasms / metabolism
  • Skin Neoplasms / pathology*
  • Transfection

Substances

  • MicroRNAs
  • Mirn199 microRNA, mouse