Induction of Mesenchymal-Epithelial Transitions in Sarcoma Cells

J Vis Exp. 2017 Apr 7:(122):55520. doi: 10.3791/55520.

Abstract

Phenotypic plasticity refers to a phenomenon in which cells transiently gain traits of another lineage. During carcinoma progression, phenotypic plasticity drives invasion, dissemination and metastasis. Indeed, while most of the studies of phenotypic plasticity have been in the context of epithelial-derived carcinomas, it turns out sarcomas, which are mesenchymal in origin, also exhibit phenotypic plasticity, with a subset of sarcomas undergoing a phenomenon that resembles a mesenchymal-epithelial transition (MET). Here, we developed a method comprising the miR-200 family and grainyhead-like 2 (GRHL2) to mimic this MET-like phenomenon observed in sarcoma patient samples.We sequentially express GRHL2 and the miR-200 family using cell transduction and transfection, respectively, to better understand the molecular underpinnings of these phenotypic transitions in sarcoma cells. Sarcoma cells expressing miR-200s and GRHL2 demonstrated enhanced epithelial characteristics in cell morphology and alteration of epithelial and mesenchymal biomarkers. Future studies using these methods can be used to better understand the phenotypic consequences of MET-like processes on sarcoma cells, such as migration, invasion, metastatic propensity, and therapy resistance.

Publication types

  • Video-Audio Media

MeSH terms

  • Cell Count
  • Cell Line, Tumor
  • DNA-Binding Proteins / metabolism*
  • Epithelial Cells / pathology
  • Epithelial-Mesenchymal Transition / physiology*
  • Gene Expression Regulation
  • Humans
  • MicroRNAs / metabolism*
  • Sarcoma / pathology*
  • Transcription Factors / metabolism*
  • Transfection

Substances

  • DNA-Binding Proteins
  • GRHL2 protein, human
  • MIRN200 microRNA, human
  • MicroRNAs
  • Transcription Factors